Team:Bielefeld-CeBiTec/Notebook/Primer
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+ | <div class="element"> | ||
+ | <table width="100%" cellspacing="10" align="center" style="background-color:transparent"> | ||
+ | <tr> | ||
+ | <td style="font-family:Georgia"><div class="element" style="margin:0px; text-align:center"><a href="#General"><img src="https://static.igem.org/mediawiki/2014/3/35/Bielefeld_CeBiTec_2014-08-25_ecoliGeneral.png" width="150px" height="150px" align="center"><br>General</a></div></td> | ||
+ | <td style="font-family:Georgia"><div class="element" style="margin:0px; text-align:center"><a href="#Module1"><img src="https://static.igem.org/mediawiki/2014/3/35/Bielefeld_CeBiTec_2014-08-14_ecoliM1.jpg" width="150px" height="150px" align="center"><br>Module 1</div></a></td> | ||
+ | <td style="font-family:Georgia"><div class="element" style="margin:0px; text-align:center"><a href="#Module2"><img src="https://static.igem.org/mediawiki/2014/6/69/Bielefeld_CeBiTec_2014-10-06_ecoliM2.jpg" width="150px" height="150px" align="center"><br>Module 2</div></a></td> | ||
+ | <td style="font-family:Georgia"><div class="element" style="margin:0px; text-align:center"><a href="#Module3"><img src="https://static.igem.org/mediawiki/2014/d/d7/Bielefeld_CeBiTec_2014-08-14_ecoliM3.jpg" width="150px" height="150px" align="center"><br>Module 3</div></a></td> | ||
+ | <td style="font-family:Georgia"><div class="element" style="margin:0px; text-align:center"><a href="#Biosafety"><img src="https://static.igem.org/mediawiki/2014/8/8a/Bielefeld_CeBiTec_2014-08-26_ecoliBiosafety.png" width="150px" height="150px" align="center"><br>Biosafety</div></a></td> | ||
+ | </tr> | ||
+ | </table> | ||
+ | </div> | ||
+ | <br> | ||
<div class="element" style="margin:10px; padding:10px; font-size:15px"> | <div class="element" style="margin:10px; padding:10px; font-size:15px"> | ||
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</tr> | </tr> | ||
<tr bgcolor="#3fb498"> | <tr bgcolor="#3fb498"> | ||
- | <th colspan="2" align="left"> | + | <th colspan="2" align="left" id="General">General</th> |
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td width="30%" | + | <td width="30%" id="VF-Primer">VF-Primer (20 bp)</td> |
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
<td width="70%">5‘-CCACCTGACGTCTAAGAAAC-3‘</td> | <td width="70%">5‘-CCACCTGACGTCTAAGAAAC-3‘</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td width="30%">VR-Primer (22 bp)</td> | + | <td width="30%" id="VR-Primer">VR-Primer (22 bp)</td> |
<td width="70%">5‘-GTATTACCGCCTTTGAGTGAGC-3‘</td> | <td width="70%">5‘-GTATTACCGCCTTTGAGTGAGC-3‘</td> | ||
</tr> | </tr> | ||
<tr bgcolor="#3fb498"> | <tr bgcolor="#3fb498"> | ||
- | <th colspan="2" align="left">Module 1</th> | + | <th colspan="2" align="left" id="Module1">Module 1</th> |
</tr> | </tr> | ||
<tr bgcolor="#D8D8D8"> | <tr bgcolor="#D8D8D8"> | ||
- | <th colspan="2" align="left">Gene synthesis</th> | + | <th colspan="2" align="left">Gene synthesis of the fumarate reductase (<i>fumA, fumBCD</i>) of <i>B. succinogenes</i></th> |
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td width="30%">pSB1C3_pre_Fum_A (43 bp)</td> | + | <td width="30%" id="pSB1C3_pre_Fum_A">pSB1C3_pre_Fum_A (43 bp)</td> |
<td width="70%">5‘-GTCTGCATTTTCTCCTCTTTCTCTAGAAGCGGCCGCGAATTCC-3‘</td> | <td width="70%">5‘-GTCTGCATTTTCTCCTCTTTCTCTAGAAGCGGCCGCGAATTCC-3‘</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td width="30%">pSB1C3_suf_Fum_A (43 bp)</td> | + | <td width="30%" id="pSB1C3_suf_Fum_A">pSB1C3_suf_Fum_A (43 bp)</td> |
<td width="70%">5‘-CCAAAGCGCCGAATGCGTAATACTAGTAGCGGCCGCTGCAGTC-3‘</td> | <td width="70%">5‘-CCAAAGCGCCGAATGCGTAATACTAGTAGCGGCCGCTGCAGTC-3‘</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td width="30%">pSB1C3_pre_Fum_BCD (42 bp)</td> | + | <td width="30%" id="pSB1C3_pre_Fum_BCD">pSB1C3_pre_Fum_BCD (42 bp)</td> |
<td width="70%">5‘-TTGGTCATTTTCTCCTCTTTCTCTAGAAGCGGCCGCGAATCC-3‘</td> | <td width="70%">5‘-TTGGTCATTTTCTCCTCTTTCTCTAGAAGCGGCCGCGAATCC-3‘</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td width="30%">pSB1C3_suf_Fum_BCD (43 bp)</td> | + | <td width="30%" id="pSB1C3_suf_Fum_BCD">pSB1C3_suf_Fum_BCD (43 bp)</td> |
<td width="70%">5‘-GGGGTGTGGCGTCGATCTAATACTAGTAGCGGCCGCTGCAGTC-3‘</td> | <td width="70%">5‘-GGGGTGTGGCGTCGATCTAATACTAGTAGCGGCCGCTGCAGTC-3‘</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td width="30%" id="FumBCD_fwd">FumBCD_fwd (24 bp)</td> | ||
+ | <td width="70%">5‘-CGGCCGCTTCTAGAGAAAGAGGAG-3‘</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td width="30%" id="FumBCD_rev">FumBCD_rev (22 bp)</td> | ||
+ | <td width="70%">5‘-CGGCCGCTACTAGTATTAGATC-3‘</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td width="30%" id="FumA_seq">FumA_seq (20 bp)</td> | ||
+ | <td width="70%">5‘-TGGAAGGCACCTTGGTTCAG-3‘</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td width="30%" id="FumB_seq">FumB_seq (20 bp)</td> | ||
+ | <td width="70%">5‘-GCTCTGACCATGGCACATCG-3‘</td> | ||
+ | </tr> | ||
+ | <tr bgcolor="#D8D8D8"> | ||
+ | <th colspan="2" align="left">Overexpression of the fumarate reductase (<i>frd</i>) of <i>E. coli</i></th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td width="30%" id="T7_frd_Ec_fwd">T7_frd_Ec_fwd (64 bp)</td> | ||
+ | <td width="70%">5‘-TAATACGACTCACTATAGGGAAAGAGGAGAAATACTAGATGCAAACCTTTCAAGCCGATCTTGC-3‘</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td width="30%" id="T7_frd_Ec_rev">T7_frd_Ec_rev (44 bp)</td> | ||
+ | <td width="70%">5‘-TGCAGCGGCCGCTACTAGTATTAGATTGTAACGACACCAATCAG-3‘</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td width="30%" id="pSB1C3_frd_pre">pSB1C3_frd_pre (45 bp)</td> | ||
+ | <td width="70%">5‘-GCAAGATCGGCTTGAAAGGTTTGCATCTAGTATTTCTCCTCTTTC-3‘</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td width="30%" id="pSB1C3_frd_suf">pSB1C3_frd_suf (47 bp)</td> | ||
+ | <td width="70%">5‘-CTGATTGGTGTCGTTACAATCTAATACTAGTAGCGGCCGCTGCAGTC-3‘</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td width="30%" id="frd_cut1_fwd">frd_cut1_fwd (20 bp)</td> | ||
+ | <td width="70%">5‘-CATACCGTTGCAGCAGAAGG-3‘</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td width="30%" id="frd_cut1_rev">frd_cut1_rev (20 bp)</td> | ||
+ | <td width="70%">5‘-CCTTCTGCTGCAACGGTATG-3‘</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td width="30%" id="frd_cut2_fwd">frd_cut2_fwd (20 bp)</td> | ||
+ | <td width="70%">5‘-GACATGGAATTTGTTCAGTA-3‘</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td width="30%" id="frd_cut2_rev">frd_cut2_rev (20 bp)</td> | ||
+ | <td width="70%">5‘-TACTGAACAAATTCCATGTC-3‘</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td width="30%" id="frd_cut3_fwd">frd_cut3_fwd (20 bp)</td> | ||
+ | <td width="70%">5‘-GAGCTGCAAGAACGCTTCAA-3‘</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td width="30%" id="frd_cut3_rev">frd_cut3_rev (20 bp)</td> | ||
+ | <td width="70%">5‘-TTGAAGCGTTCTTGCAGCTC-3‘</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <tr bgcolor="#D8D8D8"> | ||
+ | <th colspan="2" align="left">Replacement of the succinate carrier (<i>dcuB</i>) by porines (<i>oprF</i>) in the genome</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td width="30%" id="porine_fwd_FRT">porine_fwd_FRT (73 bp)</td> | ||
+ | <td width="70%">5‘-TTATAAGAACCCGTACATCGCGGCGAAGATCCAGCCGAAGACGCACGATATTGACAGCTAGCTCAGTCCTAGG-3‘</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td width="30%" id="porine_rev_FRT">porine_rev_FRT (46 bp)</td> | ||
+ | <td width="70%">5‘-CGCCCTTTAGTGAGGGTTAATTTTACTTAGCTTGGGCTTCAACCT-3‘</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td width="30%" id="pR6K_dcuB_fwd">pR6K_dcuB_fwd (46 bp)</td> | ||
+ | <td width="70%">5‘-AGGTTGAAGCCCAAGCTAAGTAAAATTAACCCTCACTAAAGGGCG-3‘</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td width="30%" id="pR6K_dcuB_rev">pR6K_dcuB_rev (73 bp)</td> | ||
+ | <td width="70%">5‘-ATGTTATTTACTATCCAACTTATCATAATACTGATATGTCTGTTTTATGGTAATACGACTCACTATAGGGCTC-3‘</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td width="30%" id="dcuB_del_kon1">dcuB_del_kon1 (20 bp)</td> | ||
+ | <td width="70%">5‘-CGCCGCATTCTGTGAATAGC-3‘</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td width="30%" id="dcuB_del_kon2">dcuB_del_kon2 (20 bp)</td> | ||
+ | <td width="70%">5‘-CCCTGGCACAAGTTACATCC-3‘</td> | ||
+ | </tr> | ||
+ | <tr bgcolor="#D8D8D8"> | ||
+ | <th colspan="2" align="left">NADPH/NADP+ sensor (<i>soxRS</i>) </th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td width="30%" id="soxR_fwd">soxR_fwd (64 bp)</td> | ||
+ | <td width="70%">5‘-TTGACAGCTAGCTCAGTCCTAGGTATTGTGCAAAGAGGAGAAAATGGAAAAGAAATTACCCCGC-3‘</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td width="30%" id="soxR_rev">soxR_rev (37 bp)</td> | ||
+ | <td width="70%">5‘-GCTTCTAGAGTTAGTTTTGTTCATCTTCCAGCAAGCG-3‘</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td width="30%" id="pSB1C3_suf_soxR">pSB1C3_suf_soxR (54 bp)</td> | ||
+ | <td width="70%">5‘-TAGGACTGAGCTAGCTGTCAATACTAGTAGCTACTAGTAGCGGCCGCTGCAGTC-3‘</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td width="30%" id="pSB1C3_pre_soxR">pSB1C3_pre_soxR (43 bp)</td> | ||
+ | <td width="70%">5‘-TGGAAGATGAACAAAACTAACTCTAGAAGCGGCCGCGAATTCC-3‘</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td width="30%" id="soxS_GFP_fwd">soxS_GFP_fwd (74 bp)</td> | ||
+ | <td width="70%">5‘-TTATACTCCCCAACAGATGAATTAACGAACTGAACACTGAAAAGAGGCAGATTTATGCGTAAAGGAGAAGAACT-3‘</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td width="30%" id="soxS_GFP_rev">soxS_GFP_rev (42 bp)</td> | ||
+ | <td width="70%">5‘-TGCAGCGGCCGCTACTAGTATATAAACGCAGAAAGGCCCACC-3‘</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td width="30%" id="pSB1C3_pre_soxS">pSB1C3_pre_soxS (78 bp)</td> | ||
+ | <td width="70%">5‘-TAATTCATCTGTTGGGGAGTATAATTCCTCAAGTTAACTTGAGGTAAAGCGATTTCTCTAGAAGCGGCCGCGAATTCC-3‘</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td width="30%" id="pSB1C3_suf_soxS">pSB1C3_suf_soxS (43 bp)</td> | ||
+ | <td width="70%">5‘-TGGGCCTTTCTGCGTTTATATACTAGTAGCGGCCGCTGCAGTC-3‘</td> | ||
</tr> | </tr> | ||
<tr bgcolor="#D8D8D8"> | <tr bgcolor="#D8D8D8"> | ||
- | <th colspan="2" align="left"> | + | <th colspan="2" align="left">Expression of the ccm cluster and cytochrome C (<i>pccH</i>)</th> |
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td width="30%">ccm_fwd (58 bp)</td> | + | <td width="30%" id="ccm_fwd">ccm_fwd (58 bp)</td> |
<td width="70%">5‘-CTCAGTCCTAGGTATTGTGCAGAGAAAGAGGAGAAAATGCTTGAAGCCAGAGAGTTAC-3‘</td> | <td width="70%">5‘-CTCAGTCCTAGGTATTGTGCAGAGAAAGAGGAGAAAATGCTTGAAGCCAGAGAGTTAC-3‘</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td width="30%">ccm_rev (47 bp)</td> | + | <td width="30%" id="ccm_rev">ccm_rev (47 bp)</td> |
<td width="70%">5‘-TGCAGCGGCCGCTACTAGTATTATTTACTCTCCTGCGGCGACAAATG-3‘</td> | <td width="70%">5‘-TGCAGCGGCCGCTACTAGTATTATTTACTCTCCTGCGGCGACAAATG-3‘</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td width="30%">pSB1C3_suf_ccm (50 bp)</td> | + | <td width="30%" id="pSB1C3_suf_ccm">pSB1C3_suf_ccm (50 bp)</td> |
<td width="70%">5‘-CATTTGTCGCCGCAGGAGAGTAAATAATACTAGTAGCGGCCGCTGCAGTC-3‘</td> | <td width="70%">5‘-CATTTGTCGCCGCAGGAGAGTAAATAATACTAGTAGCGGCCGCTGCAGTC-3‘</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td width="30%">pSB1C3_pre_ccm (28 bp)</td> | + | <td width="30%"id="pSB1C3_pre_ccm">pSB1C3_pre_ccm (28 bp)</td> |
<td width="70%">5‘-CTCTGCTAGCACAATACCTAGGACTGAG-3‘</td> | <td width="70%">5‘-CTCTGCTAGCACAATACCTAGGACTGAG-3‘</td> | ||
- | |||
- | |||
- | |||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td width="30%">pccH_fwd (55 bp)</td> | + | <td width="30%" id="pccH_fwd">pccH_fwd (55 bp)</td> |
<td width="70%">5‘-ATTCGCGGCCGCTTCTAGAGAAAGAGGAGAAAATGCAAAAAACAACTATTCTTTC-3‘</td> | <td width="70%">5‘-ATTCGCGGCCGCTTCTAGAGAAAGAGGAGAAAATGCAAAAAACAACTATTCTTTC-3‘</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td width="30%">pccH_rev (40 bp)</td> | + | <td width="30%" id="pccH_rev">pccH_rev (40 bp)</td> |
<td width="70%">5‘-TGCAGCGGCCGCTACTAGTATCAGTACGTGACCGTGATGG-3‘</td> | <td width="70%">5‘-TGCAGCGGCCGCTACTAGTATCAGTACGTGACCGTGATGG-3‘</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td width="30%">pSB1C3_pre_pccH (58 bp)</td> | + | <td width="30%" id="pSB1C3_pre_pccH">pSB1C3_pre_pccH (58 bp)</td> |
<td width="70%">5‘-GAAAGAATAGTTGTTTTTTGCATTTTCTCCTCTTTCTCTAGAAGCGGCCGCGAATTCC-3‘</td> | <td width="70%">5‘-GAAAGAATAGTTGTTTTTTGCATTTTCTCCTCTTTCTCTAGAAGCGGCCGCGAATTCC-3‘</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td width="30%">pSB1C3_suf_pccH (43 bp)</td> | + | <td width="30%" id="pSB1C3_suf_pccH">pSB1C3_suf_pccH (43 bp)</td> |
<td width="70%">5‘-CCATCACGGTCACGTACTGATACTAGTAGCGGCCGCTGCAGTC-3‘</td> | <td width="70%">5‘-CCATCACGGTCACGTACTGATACTAGTAGCGGCCGCTGCAGTC-3‘</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td width="30%" id="ccm_con1">ccm_con1 (21 bp)</td> | ||
+ | <td width="70%">5‘-ATTGGCATTGGCGGTTGGGTC-3‘</td> | ||
+ | </tr> | ||
+ | <tr bgcolor="#3fb498"> | ||
+ | <th colspan="2" align="left" id="Module2">Module 2</th> | ||
</tr> | </tr> | ||
<tr bgcolor="#D8D8D8"> | <tr bgcolor="#D8D8D8"> | ||
- | <th colspan="2" align="left"> | + | <th colspan="2" align="left">Gene synthesis of the RuBisCO of <i>Synechoccus elangatus</i> (<i>rbcLS</i>) and <i>Halothiobacillus neapolitanus</i> (<i>ccbLS</i>) and the phosphoribulokinase A (<i>prkA</i>) of <i>Synechoccus elangatus</i></th> |
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td width="30%"> | + | <td width="30%" id="pSB1C3_pre_rbcLS">pSB1C3_pre_rbcLS (44 bp)</td> |
- | <td width="70%">5‘- | + | <td width="70%">5‘-CATCTAGTATTTCTCCTCTTTCTCTAGAAGCGGCCGCGAATTCC-3‘</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td width="30%"> | + | <td width="30%" id="pSB1C3_suf_rbcLS">pSB1C3_suf_rbcLS (43 bp)</td> |
- | <td width="70%">5‘- | + | <td width="70%">5‘-ATCGTCCGGGCCGCTACTAA-TACTAGTAGCGGCCGCTGCAGTC-3‘</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td width="30%"> | + | <td width="30%" id="pSB1C3_pre_ccbLS">pSB1C3_pre_ccbLS (44 bp)</td> |
- | <td width="70%">5‘- | + | <td width="70%">5‘-CATCTAGCGGCCGCTACTAGTATTAGATCTATTTCTCCTCTTTCTCTAGAAGCGGCCGCGAATTCC-3‘</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td width="30%"> | + | <td width="30%" id="pSB1C3_suf_ccbLS">pSB1C3_suf_ccbLS (43 bp)</td> |
- | <td width="70%">5‘- | + | <td width="70%">5‘-TGGTTTACCGCGGCAACTAATACTAGTAGCGGCCGCTGCAGTC-3‘</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td width="30%"> | + | <td width="30%" id="pSB1C3_pre_prkA">pSB1C3_pre_prkA (43 bp)</td> |
- | <td width="70%">5‘- | + | <td width="70%">5‘-ACACGATCTGGCTTGCTCATCTCTAGAAGCGGCCGCGAATTCC-3‘</td> |
</tr> | </tr> | ||
- | <tr | + | <tr> |
- | + | <td width="30%" id="pSB1C3_suf_prkA">pSB1C3_suf_prkA (43 bp)</td> | |
+ | <td width="70%">5‘-CGGTTGCCGCTAGCGTTTAATACTAGTAGCGGCCGCTGCAGTC-3‘</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td width="30%" id="Hneap_seq">Hneap_seq (20 bp)</td> | ||
+ | <td width="70%">5‘-TACAGTTTGGACCGACCTTC-3‘</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td width="30%" id="Selan_seq">Selan_seq (20 bp)</td> | ||
+ | <td width="70%">5‘-GTACATGGACGACTGTATGG-3‘</td> | ||
</tr> | </tr> | ||
<tr bgcolor="#D8D8D8"> | <tr bgcolor="#D8D8D8"> | ||
- | <th colspan="2" align="left"> | + | <th colspan="2" align="left">Insert amplification of the gene synthesis of the RuBisCO of <i> S. elangatus</i> (<i>SELAN</i>)</th> |
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td width="30%"> | + | <td width="30%" id="Selan_fwd">Selan_fwd (20 bp)</td> |
- | <td width="70%">5‘- | + | <td width="70%">5‘-GGCCGCTTCTAGAGAAAGAG-3‘</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td width="30%"> | + | <td width="30%" id="Selan_rev">Selan_rev (20 bp)</td> |
- | <td width="70%">5‘- | + | <td width="70%">5‘-GCGGCCGCTACTAGTATTAG-3‘</td> |
+ | </tr> | ||
+ | <tr bgcolor="#D8D8D8"> | ||
+ | <th colspan="2" align="left">Sequencing primer for the addgene plasmid pHnCBcsoS1D</th> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td width="30%"> | + | <td width="30%" id="add_seq1">add_seq1 (20 bp)</td> |
- | <td width="70%">5‘- | + | <td width="70%">5‘-AGCGTGTCGTAGTGCGTATC-3‘</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td width="30%"> | + | <td width="30%" id="add_seq2">add_seq2 (20 bp)</td> |
- | <td width="70%">5‘- | + | <td width="70%">5‘-ACCACTTCTGGCTGCACAAC-3‘</td> |
+ | </tr> | ||
+ | <tr> | ||
+ | <td width="30%" id="add_seq3">add_seq3 (20 bp)</td> | ||
+ | <td width="70%">5‘-CTGGCAACCGGCTTGATTAC-3‘</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td width="30%" id="add_seq4">add_seq4 (20 bp)</td> | ||
+ | <td width="70%">5‘-AAATCACCCACGCCAAACCG-3‘</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td width="30%" id="add_seq5">add_seq5 (20 bp)</td> | ||
+ | <td width="70%">5‘-GATGCGATTGGCTGCATACC-3‘</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td width="30%" id="add_seq6">add_seq6 (20 bp)</td> | ||
+ | <td width="70%">5‘-GCCGTAATATCCAGCTGAAC-3‘</td> | ||
</tr> | </tr> | ||
<tr bgcolor="#D8D8D8"> | <tr bgcolor="#D8D8D8"> | ||
- | <th colspan="2" align="left"> | + | <th colspan="2" align="left">Integration of the <i>prkA</i> in the plasdmid pHnCBcsoS1D</th> |
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td width="30%">csoS2_sap_1_fwd (43)</td> | + | <td width="30%" id="pHnCBcsoS1D_fwd">pHnCBcsoS1D_fwd (44 bp)</td> |
+ | <td width="70%">5‘-CGGTTGCCGCTAGCGTTTAAGTGAGCGGATAACAATTTCAG-3‘</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td width="30%" id="pHnCBcsoS1D_rev">pHnCBcsoS1D_rev (45 bp)</td> | ||
+ | <td width="70%">5‘-CATCTAGTATTTCTCCTCTTTTCCACACATTATACGAGCCGGAAG-3‘</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td width="30%" id="prkA_pHn_rev">prkA_pHn_rev (47 bp)</td> | ||
+ | <td width="70%">5‘-CTGAAATTGTTATCCGCTCACTTAAACGCTAGCGGCAACCGGTGCTG-3‘</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td width="30%" id="prkA_pHn_fwd">prkA_pHn_fwd (53 bp)</td> | ||
+ | <td width="70%">5‘-CTTCCGGCTCGTATAATGTGTGGAAAAGAGGAGAAATACTAGATGAGCAAGCC-3‘</td> | ||
+ | </tr> | ||
+ | <tr bgcolor="#D8D8D8"> | ||
+ | <th colspan="2" align="left">Shell associated protein of the carboxysome (<i>sap</i> respectively <i>csoS2</i>)</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td width="30%" id="csoS2_sap_1_fwd">csoS2_sap_1_fwd (43)</td> | ||
<td width="70%">5‘-ATTCGCGGCCGCTTCTAGAGGTCAAGTCTGTCATTGCGCTGTG-3‘</td> | <td width="70%">5‘-ATTCGCGGCCGCTTCTAGAGGTCAAGTCTGTCATTGCGCTGTG-3‘</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td width="30%">pSB1C3_pre_sap_1 (43 bp)</td> | + | <td width="30%" id="pSB1C3_pre_sap_1">pSB1C3_pre_sap_1 (43 bp)</td> |
<td width="70%">5‘-AGCGCAATGACAGACTTGACCTCTAGAAGCGGCCGCGAATTCC-3‘</td> | <td width="70%">5‘-AGCGCAATGACAGACTTGACCTCTAGAAGCGGCCGCGAATTCC-3‘</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td width="30%">csoS2_sap_1_rev (38 bp)</td> | + | <td width="30%" id="csoS2_sap_1_rev">csoS2_sap_1_rev (38 bp)</td> |
<td width="70%">5‘-GCGCCGAACATGACTTTATCCGCTTGTGAAGGCTTTGC-3‘</td> | <td width="70%">5‘-GCGCCGAACATGACTTTATCCGCTTGTGAAGGCTTTGC-3‘</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td width="30%" id="sap_2_fwd">sap_2_fwd (22 bp)</td> | ||
+ | <td width="70%">5‘-AAGTCATGTTCGGCGCCACAGC-3‘</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td width="30%" id="sap_2_rev">sap_2_rev (20 bp)</td> | ||
+ | <td width="70%">5‘-GCCGCTACTAGTATTAACCG-3‘</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td width="30%" id="pSB1C3_suf_sap2">pSB1C3_suf_sap2 (40 bp)</td> | ||
+ | <td width="70%">5‘-CCGGCGGCGCGCGCGGTTAATACTAGTAGCGGCCGCTGCA-3‘</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td width="30%" id="seq_sap_2">seq_sap_2 (20 bp)</td> | ||
+ | <td width="70%">5‘-TGCCCGTCACCGGTAATGAG-3‘</td> | ||
</tr> | </tr> | ||
<tr bgcolor="#D8D8D8"> | <tr bgcolor="#D8D8D8"> | ||
- | <th colspan="2" align="left"> | + | <th colspan="2" align="left">Carbonic anhydrase of the carboxysome (<i>can</i>)</th> |
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td width="30%"> | + | <td width="30%" id="csoS3_can_fwd">csoS3_can_fwd (50 bp)</td> |
- | <td width="70%">5‘- | + | <td width="70%">5‘-ATTCGCGGCCGCTTCTAGAGTTAAGTAAAGTGTAACGATCATGAACACCC-3‘</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td width="30%"> | + | <td width="30%" id="pSB1C3_pre_can">pSB1C3_pre_can (46 bp)</td> |
- | <td width="70%">5‘- | + | <td width="70%">5‘-GTTCATGATCGTTACACTTTACTTAACTCTAGAAGCGGCCGCGAAT-3‘</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td width="30%"> | + | <td width="30%" id="csoS3_can_rev">csoS3_can_rev (41 bp)</td> |
- | <td width="70%">5‘- | + | <td width="70%">5‘-TGCAGCGGCCGCTACTAGTATCATGCGGATGCAACCTCTTC-3‘</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td width="30%"> | + | <td width="30%" id="pSB1C3_suf_can">pSB1C3_suf_can (46 bp)</td> |
- | <td width="70%">5‘- | + | <td width="70%">5‘-TCGAAGAGGTTGCATCCGCATGATACTAGTAGCGGCCGCTGCAGTC-3‘</td> |
</tr> | </tr> | ||
<tr bgcolor="#D8D8D8"> | <tr bgcolor="#D8D8D8"> | ||
- | <th colspan="2" align="left"> | + | <th colspan="2" align="left">Shell proteins of the carboxysome (<i>csoS4AB</i> and <i>csoS1CAB</i>)</th> |
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td width="30%">fw_pSB1C3_csoS4A (47 bp)</td> | + | <td width="30%" id="fw_pSB1C3_csoS4A">fw_pSB1C3_csoS4A (47 bp)</td> |
<td width="70%">5‘-ATTCGCGGCCGCTTCTAGAGAAATCATGCAAGTTGAGAAAACG-3‘</td> | <td width="70%">5‘-ATTCGCGGCCGCTTCTAGAGAAATCATGCAAGTTGAGAAAACG-3‘</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td width="30%">rv_csoS4A_PstI (43 bp)</td> | + | <td width="30%" id="rv_csoS4A_PstI">rv_csoS4A_PstI (43 bp)</td> |
<td width="70%">5‘-GAGGGATAAGACTTGCTTCCGGCAGCCTCTCGTGCTGCCGATG-3‘</td> | <td width="70%">5‘-GAGGGATAAGACTTGCTTCCGGCAGCCTCTCGTGCTGCCGATG-3‘</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td width="30%">fw_PstI_csoS4A (51 bp)</td> | + | <td width="30%" id="fw_PstI_csoS4A">fw_PstI_csoS4A (51 bp)</td> |
<td width="70%">5‘-TCATCGGCAGCACGAGAGGCTGCCGGAAGCAAGTCTTATCCCTCTGATTTG-3‘</td> | <td width="70%">5‘-TCATCGGCAGCACGAGAGGCTGCCGGAAGCAAGTCTTATCCCTCTGATTTG-3‘</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td width="30%">rv_SpeI_Intergen (52 bp)</td> | + | <td width="30%" id="rv_SpeI_Intergen">rv_SpeI_Intergen (52 bp)</td> |
<td width="70%">5‘-GTCGTTGCCATAACAGATCCTCTAAAACTAGGTTGAGCACGTTCAGAACGAG-3‘</td> | <td width="70%">5‘-GTCGTTGCCATAACAGATCCTCTAAAACTAGGTTGAGCACGTTCAGAACGAG-3‘</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td width="30%">fw_SpeI_Intergen (49 bp)</td> | + | <td width="30%" id="fw_SpeI_Intergen">fw_SpeI_Intergen (49 bp)</td> |
<td width="70%">5‘-TCGTTCTGAACGTGCTCAACCTAGTTTTAGAGGATCTGTTATGGCAACG-3‘</td> | <td width="70%">5‘-TCGTTCTGAACGTGCTCAACCTAGTTTTAGAGGATCTGTTATGGCAACG-3‘</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td width="30%"> | + | <td width="30%" id"=rv_csoS1_pSB1C3">rv_csoS1_pSB1C3 (50 bp)</td> |
<td width="70%">5‘-CGGACTGCAGCGGCCGCTACTAGTATTAGCTATTCAGATTTGCGATACAC-3‘</td> | <td width="70%">5‘-CGGACTGCAGCGGCCGCTACTAGTATTAGCTATTCAGATTTGCGATACAC-3‘</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td width="30%"> | + | <td width="30%" id="fw_csoS1_pSB1C3">fw_csoS1_pSB1C3 (46 bp)</td> |
<td width="70%">5‘-GTGTATCGCAAATCTGAATAGCTAATACTAGTAGCGGCCGCTGCAG-3‘</td> | <td width="70%">5‘-GTGTATCGCAAATCTGAATAGCTAATACTAGTAGCGGCCGCTGCAG-3‘</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td width="30%"> | + | <td width="30%" id="rv_pSB1C3_csoS4A">rv_pSB1C3_csoS4A (47 bp)</td> |
<td width="70%">5‘-AACGTTTTCTCAACTTGCATGATTTCTCTAGAAGCGGCCGCGAATTC-3‘</td> | <td width="70%">5‘-AACGTTTTCTCAACTTGCATGATTTCTCTAGAAGCGGCCGCGAATTC-3‘</td> | ||
</tr> | </tr> | ||
<tr bgcolor="#D8D8D8"> | <tr bgcolor="#D8D8D8"> | ||
- | <th colspan="2" align="left"> | + | <th colspan="2" align="left">Pore protein of the carboxysome (<i>csoS1D</i>)</th> |
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td width="30%">pSB1C3_pre_sRNApfkA (43 bp)</td> | + | <td width="30%" id="csoS1D_fwd">csoS1D_fwd (40 bp)</td> |
+ | <td width="70%">5‘-ATTCGCGGCCGCTTCTAGAGTGTCTAGTAGGGAAGATGCG-3‘</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td width="30%" id="csoS1D_rev">csoS1D_rev (41 bp)</td> | ||
+ | <td width="70%">5‘-TGCAGCGGCCGCTACTAGTATTAGAACCCTTCAGCGCGACG-3‘</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td width="30%" id="pSB1C3_suf_csoS1D">pSB1C3_suf_csoS1D (43bp)</td> | ||
+ | <td width="70%">5‘-GTCGCGCTGAAGGGTTCTAATACTAGTAGCGGCCGCTGCAGTC-3‘</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td width="30%" id="pSB1C3_pre_csoS1D">pSB1C3_pre_csoS1D (43 bp)</td> | ||
+ | <td width="70%">5‘-CGCATCTTCCCTACTAGACACTCTAGAAGCGGCCGCGAATTCC-3‘</td> | ||
+ | </tr> | ||
+ | <tr bgcolor="#D8D8D8"> | ||
+ | <th colspan="2" align="left">GFP fusion with the shell proteins and the shell-associated protein</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td width="30%" id="fw-csoS1A-GFP">fw-csoS1A-GFP (45 bp)</td> | ||
+ | <td width="70%">5‘-CTGCCTAAGGCGCCACAAGCCATGCGTAAAGGAGAAGAACTTTTC-3‘</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td width="30%" id="rv-csoS1B-GFP">rv-csoS1B-GFP (55 bp)</td> | ||
+ | <td width="70%">5‘-GTGAGCCGTCTTAGGAATATCTGACTTATTATTTGTATAGTTCATCCATGCCATG-3‘</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td width="30%" id="fw-GFP-csoS1B">fw-GFP-csoS1B (48 bp)</td> | ||
+ | <td width="70%">5‘-CATGGATGAACTATACAAATAATAAGTCAGATATTCCTAAGACGGCTC-3‘</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td width="30%" id="rv-GFP-csoS1A">rv-GFP-csoS1A (45 bp)</td> | ||
+ | <td width="70%">5‘-GAAAAGTTCTTCTCCTTTACGCATGGCTTGTGGCGCCTTAGGCAG-3‘</td> | ||
+ | </tr> | ||
+ | <tr bgcolor="#D8D8D8"> | ||
+ | <th colspan="2" align="left">Gluco switch using sRNA</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td width="30%" id="pSB1C3_pre_sRNApfkA">pSB1C3_pre_sRNApfkA (43 bp)</td> | ||
<td width="70%">5‘-TGTGGCTAAAAGAATTAGTACTCTAGAAGCGGCCGCGAATTCC-3‘</td> | <td width="70%">5‘-TGTGGCTAAAAGAATTAGTACTCTAGAAGCGGCCGCGAATTCC-3‘</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td width="30%">pSB1C3_suf_sRNApfkA (43 bp)</td> | + | <td width="30%" id="pSB1C3_suf_sRNApfkA">pSB1C3_suf_sRNApfkA (43 bp)</td> |
<td width="70%">5‘-TGGGCCTTTCTGCGTTTATATACTAGTAGCGGCCGCTGCAGTC-3‘</td> | <td width="70%">5‘-TGGGCCTTTCTGCGTTTATATACTAGTAGCGGCCGCTGCAGTC-3‘</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td width="30%" id="sRNA:pfkB_fwd">sRNA:pfkB_fwd (46 bp)</td> | ||
+ | <td width="70%">5‘-TACCATGCATAGATATGCAACTGTTTTCTGTTGGGCCATTGCATTG-3‘</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td width="30%" id="pSB1C3_suf_sRNApfkA">sRNA:pfkB_rev (47 bp)</td> | ||
+ | <td width="70%">5‘-ACAGTTGCATATCTATGCATGGTACTCTAGAAGCGGCCGCGAATTCC-3‘</td> | ||
</tr> | </tr> | ||
<tr bgcolor="#D8D8D8"> | <tr bgcolor="#D8D8D8"> | ||
- | <th colspan="2" align="left"> | + | <th colspan="2" align="left">Amplification of purification vector with T7, RBS and intein tag with chitin binding domain</th> |
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td width="30%">int_RBS_fw | + | <td width="30%" id="int_RBS_fw">int_RBS_fw (42 bp)</td> |
- | <td width="70%">5‘- | + | <td width="70%">5‘-CTATAGGGAAAGAGGAGAAATGCATCACGGGAGATGCGCTAG-3‘</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td width="30%">RBS_int_rev | + | <td width="30%" id="RBS_int_rev">RBS_int_rev (44 bp)</td> |
- | <td width="70%">5‘- | + | <td width="70%">5‘-CTAGCGCATCTCCCGTGATGCATTTCTCCTCTTTCCCTATAGTG-3‘</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td width="30%">RBS_rev | + | <td width="30%" id="RBS_rev">RBS_rev (22 bp)</td> |
<td width="70%">5‘-TTTCTCCTCTTTCCCTATAGTG-3‘</td> | <td width="70%">5‘-TTTCTCCTCTTTCCCTATAGTG-3‘</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td width="30%">int_fw | + | <td width="30%" id="int_fw">int_fw (22 bp)</td> |
<td width="70%">5‘-TGCATCACGGGAGATGCACTAG-3‘</td> | <td width="70%">5‘-TGCATCACGGGAGATGCACTAG-3‘</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td width="30%">int_pSB1C3_rev | + | <td width="30%" id="int_pSB1C3_rev">int_pSB1C3_rev (50 bp)</td> |
<td width="70%">5‘-GACTGCAGCGGCCGCTACTAGTATCATTGAAGCTGCCACAAGGCAGGAAC-3‘</td> | <td width="70%">5‘-GACTGCAGCGGCCGCTACTAGTATCATTGAAGCTGCCACAAGGCAGGAAC-3‘</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td width="30%">pSB1C3_int_fw | + | <td width="30%" id="pSB1C3_int_fw">pSB1C3_int_fw (46 bp)</td> |
<td width="70%">5‘-CTGCCTTGTGGCAGCTTCAATGATACTAGTAGCGGCCGCTGCAGTC-3‘</td> | <td width="70%">5‘-CTGCCTTGTGGCAGCTTCAATGATACTAGTAGCGGCCGCTGCAGTC-3‘</td> | ||
</tr> | </tr> | ||
<tr bgcolor="#D8D8D8"> | <tr bgcolor="#D8D8D8"> | ||
- | <th colspan="2" align="left">SBPase (glpX)</th> | + | <th colspan="2" align="left">Expression of the SBPase (<i>glpX</i>) for the calvin cycle and for purification</th> |
+ | </tr> | ||
+ | <tr> | ||
+ | <td width="30%" id="fw-pSB1C3-BBa_B0034-SBPase">fw-pSB1C3-BBa_B0034-SBPase (58 bp)</td> | ||
+ | <td width="70%">5‘-GAATTCGCGGCCGCTTCTAGAGAAAGAGGAGAAAATGAGGGAATTGAAAAGCGAAAAG-3‘</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td width="30%" id="rv-pSB1C3-SBPase">rv-pSB1C3-SBPase (56 bp)</td> | ||
+ | <td width="70%">5‘-GACTGCAGCGGCCGCTACTAGTATCAATGATAAGCTTCAATAAATTGGTATTTTAC-3‘</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td width="30%" id="fw-SBPase-Schnittstelle">fw-SBPase-Schnittstelle (56 bp)</td> | ||
+ | <td width="70%">5‘-AAAAGCAAAAGGCTGCGTAAATTTGGACGCATCTTTAACAGAAAATATGAAATCAG-3‘</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td width="30%" id="rv-SBPase-Schnittstelle">rv-SBPase-Schnittstelle (46 bp)</td> | ||
+ | <td width="70%">5‘-TTTTCTGTTAAAGATGCGTCCAAATTTACGCAGCCTTTTGCTTTTG-3‘</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td width="30%" id="fw-SBPase-pSB1C3">fw-SBPase-pSB1C3 (57 bp)</td> | ||
+ | <td width="70%">5‘-AATTTATTGAAGCTTATCATTGATACTAGTAGCGGCCGCTGCAGTCCGGCAAAAAAG-3‘</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td width="30%" id="rv-SBPase-BBa_B0034-pSB1C3">rv-SBPase-BBa_B0034-pSB1C3 (65 bp)</td> | ||
+ | <td width="70%">5‘-TTTTCGCTTTTCAATTCCCTCATTTTCTCCTCTTTCTCTAGAAGCGGCCGCGAATTCCAGAAATC-3‘</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td width="30%">SBPase_purify_fw (45 bp)</td> | + | <td width="30%" id="SBPase_purify_fw">SBPase_purify_fw (45 bp)</td> |
<td width="70%">5‘-CTATAGGGAAAGAGGAGAAATATGAGGGAATTGAAAAGCGAAAAG-3‘</td> | <td width="70%">5‘-CTATAGGGAAAGAGGAGAAATATGAGGGAATTGAAAAGCGAAAAG-3‘</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td width="30%">SBPase_purify_rev (53 bp)</td> | + | <td width="30%" id="SBPase_purfiy_rev">SBPase_purify_rev (53 bp)</td> |
<td width="70%">5‘-CTAGTGCATCTCCCGTGATGCACGTAAAATACCAATTTATTGAAGCTTATCAT-3‘</td> | <td width="70%">5‘-CTAGTGCATCTCCCGTGATGCACGTAAAATACCAATTTATTGAAGCTTATCAT-3‘</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td width="30%">fw_pSB1_SBPase (58 bp)</td> | + | <td width="30%" id="fw_pSB1_SBPase">fw_pSB1_SBPase (58 bp)</td> |
<td width="70%">5‘-GAATTCGCGGCCGCTTCTAGAGAAAGAGGAGAAAATGAGGGAATTGAAAAGCGAAAAG-3‘</td> | <td width="70%">5‘-GAATTCGCGGCCGCTTCTAGAGAAAGAGGAGAAAATGAGGGAATTGAAAAGCGAAAAG-3‘</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td width="30%">rv_pSB1_SBPase (56 bp)</td> | + | <td width="30%" id="rv_pSB1_SBPase">rv_pSB1_SBPase (56 bp)</td> |
<td width="70%">5‘-GACTGCAGCGGCCGCTACTAGTATCAATGATAAGCTTCAATAAATTGGTATTTTAC-3‘</td> | <td width="70%">5‘-GACTGCAGCGGCCGCTACTAGTATCAATGATAAGCTTCAATAAATTGGTATTTTAC-3‘</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td width="30%">fw_SBPase_XbaI (56 bp)</td> | + | <td width="30%" id="fw_SBPase_XbaI">fw_SBPase_XbaI (56 bp)</td> |
<td width="70%">5‘-AAAAGCAAAAGGCTGCGTAAATTTGGACGCATCTTTAACAGAAAATATGAAATCAG-3‘</td> | <td width="70%">5‘-AAAAGCAAAAGGCTGCGTAAATTTGGACGCATCTTTAACAGAAAATATGAAATCAG-3‘</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td width="30%">rv_SBPase_XbaI (46 bp)</td> | + | <td width="30%" id="rv_SBPase_XbaI">rv_SBPase_XbaI (46 bp)</td> |
<td width="70%">5‘-TTTTCTGTTAAAGATGCGTCCAAATTTACGCAGCCTTTTGCTTTTG-3‘</td> | <td width="70%">5‘-TTTTCTGTTAAAGATGCGTCCAAATTTACGCAGCCTTTTGCTTTTG-3‘</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td width="30%">fw_SBPase_pSB1C3 (57 bp)</td> | + | <td width="30%" id="fw_SBPase_pSB1C3">fw_SBPase_pSB1C3 (57 bp)</td> |
<td width="70%">5‘-AATTTATTGAAGCTTATCATTGATACTAGTAGCGGCCGCTGCAGTCCGGCAAAAAAG-3‘</td> | <td width="70%">5‘-AATTTATTGAAGCTTATCATTGATACTAGTAGCGGCCGCTGCAGTCCGGCAAAAAAG-3‘</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td width="30%">rv_SBPase_pSB1 (65 bp)</td> | + | <td width="30%" id="rv_SBPase_pSB1">rv_SBPase_pSB1 (65 bp)</td> |
<td width="70%">5‘-TTTTCGCTTTTCAATTCCCTCATTTTCTCCTCTTTCTCTAGAAGCGGCCGCGAATTCCAGAAATC-3‘</td> | <td width="70%">5‘-TTTTCGCTTTTCAATTCCCTCATTTTCTCCTCTTTCTCTAGAAGCGGCCGCGAATTCCAGAAATC-3‘</td> | ||
</tr> | </tr> | ||
<tr bgcolor="#D8D8D8"> | <tr bgcolor="#D8D8D8"> | ||
- | <th colspan="2" align="left">fbaA</th> | + | <th colspan="2" align="left">Amplification of <i>E. coli</i> fructose bisphosphate aldolase II (fbaA) for purification</th> |
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td width="30%">fbaA_purify_fw (50 bp)</td> | + | <td width="30%" id="fbaA_purify_fw">fbaA_purify_fw (50 bp)</td> |
<td width="70%">5‘-CTATAGGGAAAGAGGAGAAATATGTCTAAGATTTTTGATTTCGTAAAACC-3‘</td> | <td width="70%">5‘-CTATAGGGAAAGAGGAGAAATATGTCTAAGATTTTTGATTTCGTAAAACC-3‘</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td width="30%">fbaA_purify_rev (41 bp)</td> | + | <td width="30%" id="fbaA_purify_rev">fbaA_purify_rev (41 bp)</td> |
<td width="70%">5‘-CTAGTGCATCTCCCGTGATGCACAGAACGTCGATCGCGTTC-3‘</td> | <td width="70%">5‘-CTAGTGCATCTCCCGTGATGCACAGAACGTCGATCGCGTTC-3‘</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td width="30%">fbaA_PstI_fw (42 bp)</td> | + | <td width="30%" id="fbaA_PstI_fw">fbaA_PstI_fw (42 bp)</td> |
<td width="70%">5‘-CAAAGCGAACGAAGCTTATCTGCAAGGTCAGCTGGGTAACCC-3‘</td> | <td width="70%">5‘-CAAAGCGAACGAAGCTTATCTGCAAGGTCAGCTGGGTAACCC-3‘</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td width="30%">fbaA_PstI_rev (43 bp)</td> | + | <td width="30%" id="fbaA_PstI_rev">fbaA_PstI_rev (43 bp)</td> |
<td width="70%">5‘-CTTTCGGGTTACCCAGCTGACCTTGCAGATAAGCTTCGTTCGC-3‘</td> | <td width="70%">5‘-CTTTCGGGTTACCCAGCTGACCTTGCAGATAAGCTTCGTTCGC-3‘</td> | ||
</tr> | </tr> | ||
<tr bgcolor="#D8D8D8"> | <tr bgcolor="#D8D8D8"> | ||
- | <th colspan="2" align="left">tktA</th> | + | <th colspan="2" align="left">Amplification of <i>E. coli</i> transketolase I (tktA) for purification</th> |
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td width="30%">tktA_purify_fw (43 bp)</td> | + | <td width="30%" id="tktA_purify_fw">tktA_purify_fw (43 bp)</td> |
<td width="70%">5‘-CTATAGGGAAAGAGGAGAAATATGTCCTCACGTAAAGAGCTTG-3‘</td> | <td width="70%">5‘-CTATAGGGAAAGAGGAGAAATATGTCCTCACGTAAAGAGCTTG-3‘</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td width="30%">tktA_purify_rev (43 bp)</td> | + | <td width="30%" id="tktA_purify_rev">tktA_purify_rev (43 bp)</td> |
<td width="70%">5‘-CTAGTGCATCTCCCGTGATGCACAGCAGTTCTTTTGCTTTCGC-3‘</td> | <td width="70%">5‘-CTAGTGCATCTCCCGTGATGCACAGCAGTTCTTTTGCTTTCGC-3‘</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td width="30%">tktA_PstI_fw (41 bp)</td> | + | <td width="30%" id="tktA_PstI_fw">tktA_PstI_fw (41 bp)</td> |
<td width="70%">5‘-GTTCGGTCCGCTGTTGCCGGAACTCCTCGGCGGTTCTGCTG-3‘</td> | <td width="70%">5‘-GTTCGGTCCGCTGTTGCCGGAACTCCTCGGCGGTTCTGCTG-3‘</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td width="30%">tktA_PstI_rev (42 bp)</td> | + | <td width="30%" id="tktA_PstI_rev">tktA_PstI_rev (42 bp)</td> |
<td width="70%">5‘-CAGGTCAGCAGAACCGCCGAGGAGTTCCGGCAACAGCGGACC-3‘</td> | <td width="70%">5‘-CAGGTCAGCAGAACCGCCGAGGAGTTCCGGCAACAGCGGACC-3‘</td> | ||
</tr> | </tr> | ||
<tr bgcolor="#D8D8D8"> | <tr bgcolor="#D8D8D8"> | ||
- | <th colspan="2" align="left"> | + | <th colspan="2" align="left">Amplification of <i>C. glutamicum</i> phosphoglycerate kinase (pgk) for purification</th> |
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td width="30%">cg_pgk_purify_fw (42 bp)</td> | + | <td width="30%" id="cg_pgk_purify_fw">cg_pgk_purify_fw (42 bp)</td> |
<td width="70%">5‘-CTATAGGGAAAGAGGAGAAATATGGCTGTTAAGACCCTCAAG-3‘</td> | <td width="70%">5‘-CTATAGGGAAAGAGGAGAAATATGGCTGTTAAGACCCTCAAG-3‘</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td width="30%">cg_pgk_purify_rev (42 bp)</td> | + | <td width="30%" id="cg_pgk_purify_rev">cg_pgk_purify_rev (42 bp)</td> |
<td width="70%">5‘-CTAGTGCATCTCCCGTGATGCACTGAGCGAGAATTGCAACGC-3‘</td> | <td width="70%">5‘-CTAGTGCATCTCCCGTGATGCACTGAGCGAGAATTGCAACGC-3‘</td> | ||
</tr> | </tr> | ||
<tr bgcolor="#D8D8D8"> | <tr bgcolor="#D8D8D8"> | ||
- | <th colspan="2" align="left">gapA</th> | + | <th colspan="2" align="left">Amplification of <i>E. coli</i> glyceraldehyde 3-phosphate dehydrogenase (gapA) for purification</th> |
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td width="30%">gapA_purify_fw (47 bp)</td> | + | <td width="30%" id="gapA_purify_fw">gapA_purify_fw (47 bp)</td> |
<td width="70%">5‘-CTATAGGGAAAGAGGAGAAATATGACTATCAAAGTAGGTATCAACGG-3‘</td> | <td width="70%">5‘-CTATAGGGAAAGAGGAGAAATATGACTATCAAAGTAGGTATCAACGG-3‘</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td width="30%">gapA_purify_rev (43 bp)</td> | + | <td width="30%" id="gapA_purify_rev">gapA_purify_rev (43 bp)</td> |
<td width="70%">5‘-CTAGTGCATCTCCCGTGATGCATTTGGAGATGTGAGCGATCAG-3‘</td> | <td width="70%">5‘-CTAGTGCATCTCCCGTGATGCATTTGGAGATGTGAGCGATCAG-3‘</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td width="30%">gapA_NotI_fw (44 bp)</td> | + | <td width="30%" id="gapA_NotI_fw">gapA_NotI_fw (44 bp)</td> |
<td width="70%">5‘-CTCACAAAGACTGGCGCGGCGGCCGTGGCGCTTCCCAGAACATC-3‘</td> | <td width="70%">5‘-CTCACAAAGACTGGCGCGGCGGCCGTGGCGCTTCCCAGAACATC-3‘</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td width="30%">gapA_NotI_rev (43 bp)</td> | + | <td width="30%" id="gapA_NotI_rev">gapA_NotI_rev (43 bp)</td> |
<td width="70%">5‘-GATGATGTTCTGGGAAGCGCCACGGCCGCCGCGCCAGTCTTTG-3‘</td> | <td width="70%">5‘-GATGATGTTCTGGGAAGCGCCACGGCCGCCGCGCCAGTCTTTG-3‘</td> | ||
</tr> | </tr> | ||
<tr bgcolor="#D8D8D8"> | <tr bgcolor="#D8D8D8"> | ||
- | <th colspan="2" align="left">prkA</th> | + | <th colspan="2" align="left">Amplification of <i>S. elongatus</i> phosphoribulokinase (prkA) for purification</th> |
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td width="30%">prkA_purify_fw (40 bp)</td> | + | <td width="30%" id="prkA_purify_fw">prkA_purify_fw (40 bp)</td> |
<td width="70%">5‘-CTATAGGGAAAGAGGAGAAATATGAGCAAGCCAGATCGTG-3‘</td> | <td width="70%">5‘-CTATAGGGAAAGAGGAGAAATATGAGCAAGCCAGATCGTG-3‘</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td width="30%">prkA_purify_rev (42 bp)</td> | + | <td width="30%" id="prkA_purify_rev">prkA_purify_rev (42 bp)</td> |
<td width="70%">5‘-CTAGTGCATCTCCCGTGATGCAAACGCTAGCGGCAACCGGTG-3‘</td> | <td width="70%">5‘-CTAGTGCATCTCCCGTGATGCAAACGCTAGCGGCAACCGGTG-3‘</td> | ||
</tr> | </tr> | ||
<tr bgcolor="#D8D8D8"> | <tr bgcolor="#D8D8D8"> | ||
- | <th colspan="2" align="left">pykF</th> | + | <th colspan="2" align="left">Amplification of <i>E. coli</i> pyruvate kinase (pykF) for purification</th> |
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td width="30%">pykF_purify_fw (47 bp)</td> | + | <td width="30%" id="pykF_purify_fw">pykF_purify_fw (47 bp)</td> |
<td width="70%">5‘-CTATAGGGAAAGAGGAGAAATATGAAAAAGACCAAAATTGTTTGCAC-3‘</td> | <td width="70%">5‘-CTATAGGGAAAGAGGAGAAATATGAAAAAGACCAAAATTGTTTGCAC-3‘</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td width="30%">pykF_purify_rev (42 bp)</td> | + | <td width="30%" id="pykF_purify_rev">pykF_purify_rev (42 bp)</td> |
<td width="70%">5‘-CTAGTGCATCTCCCGTGATGCACAGGACGTGAACAGATGCGG-3‘</td> | <td width="70%">5‘-CTAGTGCATCTCCCGTGATGCACAGGACGTGAACAGATGCGG-3‘</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td width="30%">pykF_PstI_fw (48 bp)</td> | + | <td width="30%" id="pykF_PstI_fw">pykF_PstI_fw (48 bp)</td> |
<td width="70%">5‘-ACCGTCTGGGTAAAGAACTGGCTCTGCAAAGCGGTCTGGCACACAAAG-3‘</td> | <td width="70%">5‘-ACCGTCTGGGTAAAGAACTGGCTCTGCAAAGCGGTCTGGCACACAAAG-3‘</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td width="30%">pykF_PstI_rev (43 bp)</td> | + | <td width="30%" id="pykF_PstI_rev">pykF_PstI_rev (43 bp)</td> |
<td width="70%">5‘-CACCTTTGTGTGCCAGACCGCTTTGCAGAGCCAGTTCTTTACC-3‘</td> | <td width="70%">5‘-CACCTTTGTGTGCCAGACCGCTTTGCAGAGCCAGTTCTTTACC-3‘</td> | ||
</tr> | </tr> | ||
<tr bgcolor="#D8D8D8"> | <tr bgcolor="#D8D8D8"> | ||
- | <th colspan="2" align="left">ldhA</th> | + | <th colspan="2" align="left">Amplification of <i>E. coli</i> D-lactate dehydrogenase (ldhA) for purification</th> |
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td width="30%">ldhA_purify_fw (44 bp)</td> | + | <td width="30%" id="ldhA_purify_fw">ldhA_purify_fw (44 bp)</td> |
<td width="70%">5‘-CTATAGGGAAAGAGGAGAAATATGAAACTCGCCGTTTATAGCAC-3‘</td> | <td width="70%">5‘-CTATAGGGAAAGAGGAGAAATATGAAACTCGCCGTTTATAGCAC-3‘</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td width="30%">ldhA_purify_rev (42 bp)</td> | + | <td width="30%" id="ldhA_purify_rev">ldhA_purify_rev (42 bp)</td> |
<td width="70%">5‘-CTAGTGCATCTCCCGTGATGCAAACCAGTTCGTTCGGGCAGG-3‘</td> | <td width="70%">5‘-CTAGTGCATCTCCCGTGATGCAAACCAGTTCGTTCGGGCAGG-3‘</td> | ||
</tr> | </tr> | ||
<tr bgcolor="#D8D8D8"> | <tr bgcolor="#D8D8D8"> | ||
- | <th colspan="2" align="left">glk</th> | + | <th colspan="2" align="left">Amplification of <i>E. coli</i> glucokinase (glk) for purification</th> |
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td width="30%">glk_purify_fw (44 bp)</td> | + | <td width="30%" id="glk_purify_fw">glk_purify_fw (44 bp)</td> |
<td width="70%">5‘-CTATAGGGAAAGAGGAGAAATATGACAAAGTATGCATTAGTCGG-3‘</td> | <td width="70%">5‘-CTATAGGGAAAGAGGAGAAATATGACAAAGTATGCATTAGTCGG-3‘</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td width="30%">glk_purify_rev (43 bp)</td> | + | <td width="30%" id="glk_purify_rev">glk_purify_rev (43 bp)</td> |
<td width="70%">5‘-CTAGTGCATCTCCCGTGATGCACAGAATGTGACCTAAGGTCTG-3‘</td> | <td width="70%">5‘-CTAGTGCATCTCCCGTGATGCACAGAATGTGACCTAAGGTCTG-3‘</td> | ||
</tr> | </tr> | ||
<tr bgcolor="#D8D8D8"> | <tr bgcolor="#D8D8D8"> | ||
- | <th colspan="2" align="left">zwf</th> | + | <th colspan="2" align="left">Amplification of <i>E. coli</i> glucose 6-phosphate-1-dehydrogenase (zwf) for purification</th> |
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td width="30%">zwf_purify_fw (41 bp)</td> | + | <td width="30%" id="zwf_purify_fw">zwf_purify_fw (41 bp)</td> |
<td width="70%">5‘-CTATAGGGAAAGAGGAGAAATATGGCGGTAACGCAAACAGC-3‘</td> | <td width="70%">5‘-CTATAGGGAAAGAGGAGAAATATGGCGGTAACGCAAACAGC-3‘</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td width="30%">zwf_purify_rev (45 bp)</td> | + | <td width="30%" id="zwf_purify_rev">zwf_purify_rev (45 bp)</td> |
<td width="70%">5‘-CTAGTGCATCTCCCGTGATGCACTCAAACTCATTCCAGGAACGAC-3‘</td> | <td width="70%">5‘-CTAGTGCATCTCCCGTGATGCACTCAAACTCATTCCAGGAACGAC-3‘</td> | ||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
</tr> | </tr> | ||
<tr bgcolor="#3fb498"> | <tr bgcolor="#3fb498"> | ||
- | <th colspan="2" align="left">Module 3</th> | + | <th colspan="2" align="left" id="Module3">Module 3</th> |
</tr> | </tr> | ||
<tr bgcolor="#D8D8D8"> | <tr bgcolor="#D8D8D8"> | ||
- | <th colspan="2" align="left">Backbone</th> | + | <th colspan="2" align="left">Backbone for the isobutanol plasmid</th> |
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td width="30%"> | + | <td width="30%" id="fw_kivD_pSB1C3">fw_kivD_pSB1C3 (53 bp)</td> |
<td width="70%">5‘-GCAAACTATTTGCTGAACAAAATAAATCATAATACTAGTAGCGGCCGCTGCAG-3‘</td> | <td width="70%">5‘-GCAAACTATTTGCTGAACAAAATAAATCATAATACTAGTAGCGGCCGCTGCAG-3‘</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td width="30%"> | + | <td width="30%" id="rv_alsS_pSB1C3">rv_alsS_pSB1C3 (65 bp)</td> |
<td width="70%">5‘-TTTTTGTTCTTTTGTTGCTTTTGTCAACATTTTCTCCTCTTTCTCTAGAAGCGGCCGCGAATTC-3‘</td> | <td width="70%">5‘-TTTTTGTTCTTTTGTTGCTTTTGTCAACATTTTCTCCTCTTTCTCTAGAAGCGGCCGCGAATTC-3‘</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td width="30%">fw_adhA_pSB1C3 (51 bp)</td> | + | <td width="30%" id="fw_adhA_pSB1C3">fw_adhA_pSB1C3 (51 bp)</td> |
<td width="70%">5‘-GGCCGAATGGTCATTGATTTTACTAAATAATACTAGTAGCGGCCGCTGCAG-3‘</td> | <td width="70%">5‘-GGCCGAATGGTCATTGATTTTACTAAATAATACTAGTAGCGGCCGCTGCAG-3‘</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td width="30%"> | + | <td width="30%" id="fw_adhA_pSB1C3">fw_adhA_pSB1C3 (42 bp)</td> |
<td width="70%">5‘-CATTGATTTTACTAAATAATACTAGTAGCGGCCGCTGCAGTC-3‘</td> | <td width="70%">5‘-CATTGATTTTACTAAATAATACTAGTAGCGGCCGCTGCAGTC-3‘</td> | ||
</tr> | </tr> | ||
Line 470: | Line 717: | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td width="30%"> | + | <td width="30%"id="fw_pSB1C3_alsS">fw_pSB1C3_alsS (79 bp)</td> |
<td width="70%">5‘-TTCGCTAAGGATGATTTCTGGAATTCGCGGCCGCTTCTAGAGAAAGAGGAGAAAATGTTGACAAAAGCAACAAAAGAAC-3‘</td> | <td width="70%">5‘-TTCGCTAAGGATGATTTCTGGAATTCGCGGCCGCTTCTAGAGAAAGAGGAGAAAATGTTGACAAAAGCAACAAAAGAAC-3‘</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td width="30%"> | + | <td width="30%" id="rv_ilvC_alsS">rv_ilvC_alsS (64 bp)</td> |
<td width="70%">5‘-GATTCAGTGTATTGAAGTAGTTAGCCATTTTCTCCTCTTTCTAGAGAGCTTTCGTTTTCATGAG-3‘</td> | <td width="70%">5‘-GATTCAGTGTATTGAAGTAGTTAGCCATTTTCTCCTCTTTCTAGAGAGCTTTCGTTTTCATGAG-3‘</td> | ||
</tr> | </tr> | ||
Line 481: | Line 728: | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td width="30%"> | + | <td width="30%" id="fw_alsS_ilvC">fw_alsS_ilvC (65 bp)</td> |
<td width="70%">5‘-CTCATGAAAACGAAAGCTCTCTAGAAAGAGGAGAAAATGGCTAACTACTTCAATACACTGAATC-3‘</td> | <td width="70%">5‘-CTCATGAAAACGAAAGCTCTCTAGAAAGAGGAGAAAATGGCTAACTACTTCAATACACTGAATC-3‘</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td width="30%"> | + | <td width="30%" id="rev_ilvD_ilvC">rev_ilvD_ilvC (56 bp)</td> |
<td width="70%">5‘-GTGGCGGAACGGTACTTAGGCATTTTCTCCTCTTTTTAACCCGCAACAGCAATACG-3‘</td> | <td width="70%">5‘-GTGGCGGAACGGTACTTAGGCATTTTCTCCTCTTTTTAACCCGCAACAGCAATACG-3‘</td> | ||
</tr> | </tr> | ||
Line 492: | Line 739: | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td width="30%"> | + | <td width="30%" id="fw_ilvC_ilvD">fw_ilvC_ilvD (57 bp)</td> |
<td width="70%">5‘-CGTATTGCTGTTGCGGGTTAAAAAGAGGAGAAAATGCCTAAGTACCGTTCCGCCAC-3‘</td> | <td width="70%">5‘-CGTATTGCTGTTGCGGGTTAAAAAGAGGAGAAAATGCCTAAGTACCGTTCCGCCAC-3‘</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td width="30%"> | + | <td width="30%" id="rv_kivD_ilvD">rv_kivD_ilvD (67 bp)</td> |
<td width="70%">5‘-GTCTAATAGGTAATCTCCTACTGTATACATTTTCTCCTCTTTTTAACCCCCCAGTTTCGATTTATC-3‘</td> | <td width="70%">5‘-GTCTAATAGGTAATCTCCTACTGTATACATTTTCTCCTCTTTTTAACCCCCCAGTTTCGATTTATC-3‘</td> | ||
</tr> | </tr> | ||
Line 503: | Line 750: | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td width="30%"> | + | <td width="30%" id="fw_ilvD_kivD">fw_ilvD_kivD (67 bp)</td> |
<td width="70%">5‘-GATAAATCGAAACTGGGGGGTTAAAAAGAGGAGAAAATGTATACAGTAGGAGATTACCTATTAGAC-3‘</td> | <td width="70%">5‘-GATAAATCGAAACTGGGGGGTTAAAAAGAGGAGAAAATGTATACAGTAGGAGATTACCTATTAGAC-3‘</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td width="30%"> | + | <td width="30%" id="rv_pSB1C3_kivD">rv_pSB1C3_kivD (74 bp)</td> |
<td width="70%">5‘-ACCTTGCCCTTTTTTGCCGGACTGCAGCGGCCGCTACTAGTATTATGATTTATTTTGTTCAGCAAATAGTTTGC-3‘</td> | <td width="70%">5‘-ACCTTGCCCTTTTTTGCCGGACTGCAGCGGCCGCTACTAGTATTATGATTTATTTTGTTCAGCAAATAGTTTGC-3‘</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td width="30%"> | + | <td width="30%" id="rv_adhA_kivD">rv_adhA_kivD (69 bp)</td> |
<td width="70%">5‘-GTGTCTTACTACTGCTGCTTTCATTTTCTCCTCTTTTTATGATTTATTTTGTTCAGCAAATAGTTTGC-3‘</td> | <td width="70%">5‘-GTGTCTTACTACTGCTGCTTTCATTTTCTCCTCTTTTTATGATTTATTTTGTTCAGCAAATAGTTTGC-3‘</td> | ||
</tr> | </tr> | ||
<tr bgcolor="#D8D8D8"> | <tr bgcolor="#D8D8D8"> | ||
- | <th colspan="2" align="left">adhA</th> | + | <th colspan="2" align="left">Alcohol dehydrogenase (<i>adhA</i>) of <i>Lactococcus lactis<i></th> |
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td width="30%"> | + | <td width="30%" id="fw_kivD_adhA">fw_kivD_adhA (69 bp)</td> |
<td width="70%">5‘-GCAAACTATTTGCTGAACAAAATAAATCATAAAAAGAGGAGAAAATGAAAGCAGCAGTAGTAAGACAC-3‘</td> | <td width="70%">5‘-GCAAACTATTTGCTGAACAAAATAAATCATAAAAAGAGGAGAAAATGAAAGCAGCAGTAGTAAGACAC-3‘</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td width="30%"> | + | <td width="30%" id="rev_pSB1C3_adhA">rev_pSB1C3_adhA (80 bp)</td> |
<td width="70%">5‘-GTGACACCTTGCCCTTTTTTGCCGGACTGCAGCGGCCGCTACTAGTATTATTTAGTAAAATCAATGACCATTCGGCCTTC-3‘</td> | <td width="70%">5‘-GTGACACCTTGCCCTTTTTTGCCGGACTGCAGCGGCCGCTACTAGTATTATTTAGTAAAATCAATGACCATTCGGCCTTC-3‘</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td width="30%">fw_pSB1C3_adhA (66 bp)</td> | + | <td width="30%" id="fw_pSB1C3_adhA">fw_pSB1C3_adhA (66 bp)</td> |
<td width="70%">5‘-GATTTCTGGAATTCGCGGCCGCTTCTAGAGAAAGAGGAGAAAATGAAAGCAGCAGTAGTAAGACAC-3‘</td> | <td width="70%">5‘-GATTTCTGGAATTCGCGGCCGCTTCTAGAGAAAGAGGAGAAAATGAAAGCAGCAGTAGTAAGACAC-3‘</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td width="30%">rev_adhA_pSB1C3 (56 bp)</td> | + | <td width="30%" id="rev_adhA_pSB1C3">rev_adhA_pSB1C3 (56 bp)</td> |
<td width="70%">5‘-GTCTTACTACTGCTGCTTTCATTTTCTCCTCTTTCTCTAGAAGCGGCCGCGAATTC-3‘</td> | <td width="70%">5‘-GTCTTACTACTGCTGCTTTCATTTTCTCCTCTTTCTCTAGAAGCGGCCGCGAATTC-3‘</td> | ||
</tr> | </tr> | ||
+ | <tr> | ||
+ | <td width="30%" id="fw-pSB1C3-BBa_B0034-adhaA">fw-pSB1C3-BBa_B0034-adhaA (66 bp)</td> | ||
+ | <td width="70%">5‘-GATTTCTGGAATTCGCGGCCGCTTCTAGAGAAAGAGGAGAAAATGAAAGCAGCAGTAGTAAGACAC-3‘</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td width="30%" id="rev-adhA-BBa_B0034-pSB1C3">rev-adhA-BBa_B0034-pSB1C3-adhaA (56 bp)</td> | ||
+ | <td width="70%">5‘-GTCTTACTACTGCTGCTTTCATTTTCTCCTCTTTCTCTAGAAGCGGCCGCGAATTC-3‘</td> | ||
+ | </tr> | ||
+ | <tr bgcolor="#D8D8D8"> | ||
+ | <th colspan="2" align="left">Illegal restriction site between alsS and ilvC</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td width="30%" id="rv_ilvC_alsS-new">rv_ilvC_alsS-new (72 bp)</td> | ||
+ | <td width="70%">5‘-CAGTGTATTGAAGTAGTTAGCCATTTTCTCCTCTTTTTAGACAGTTTCCGTTTTCATGAGTTCCCCAAATTC-3‘</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td width="30%" id="fw_alsS_ilvC-new">fw_alsS_ilvC-new (62 bp)</td> | ||
+ | <td width="70%">5‘-CATGAAAACGGAAACTGTCTAAAAAGAGGAGAAAATGGCTAACTACTTCAATACACTGAATC-3‘</td> | ||
+ | </tr> | ||
+ | <tr bgcolor="#3fb498"> | ||
+ | <th colspan="2" align="left" id="Biosafety">Biosafety and antibiotic-free selection</th> | ||
+ | </tr> | ||
+ | <tr bgcolor="#D8D8D8"> | ||
+ | <th colspan="2" align="left" id="Deletion_alanine_racemase">Deletion of alanine racemases (<i>alr</i> and <i>dadX</i>)</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td width="30%" id="alr_fwd_redET">alr_fwd_redET (72 bp)</td> | ||
+ | <td width="70%">5‘-AATTAGGTAATTAAAGCAAACACTTATCAAGGAACACAAATGCAAGCGGCAATTAACCCTCACTAAAGGGCG-3‘</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td width="30%" id="alr_rev_redEt">alr_rev_redEt (73 bp)</td> | ||
+ | <td width="70%">5‘-TTTTATTGGCCGGATAAGGCATTCACGCCGCATCCGGCACAGACAATCAATAATACGACTCACTATAGGGCTC-3‘</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td width="30%" id="dadX_fwd_redET">dadX_fwd_redET (72 bp)</td> | ||
+ | <td width="70%">5‘-CGTCCGGGCCATTTACATGGCGCACACAGCTAAGGAAACGAGATGACCCAAATTAACCCTCACTAAAGGGCG-3‘</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td width="30%" id="dadX_rev_redET">dadX_rev_redET (73 bp)</td> | ||
+ | <td width="70%">5‘-TGATTTTTTTGCACCCAGAAGACGTTGCCTCCGATCCGGCTTACAACAAGTAATACGACTCACTATAGGGCTC-3‘</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td width="30%" id="alr_Ec_control1">alr_Ec_control1 (20 bp)</td> | ||
+ | <td width="70%">5‘-GCTGGAGATGCCATCAGAAC-3‘</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td width="30%" id="alr_Ec_control2">alr_Ec_control2 (20 bp)</td> | ||
+ | <td width="70%">5‘-CCGGCGAATATTGCTACGTG-3‘</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td width="30%" id="dadX_Ec_control1">dadX_Ec_control1 (20 bp)</td> | ||
+ | <td width="70%">5‘-GCTTTAATACGCCCGTTGAC-3‘</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td width="30%" id="dadX_Ec_control2">dadX_Ec_control2 (20 bp)</td> | ||
+ | <td width="70%">5‘-CTGGATCAACGCTTCTTTGG-3‘</td> | ||
+ | </tr> | ||
+ | <tr bgcolor="#D8D8D8"> | ||
+ | <th colspan="2" align="left">Replacement of the chloramphenicol resistance (Cm) by the konstitutive alanine racemase <i>alr</i></th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td width="30%" id="pSB1C3_Cm_del_fwd">pSB1C3_Cm_del_fwd (40 bp)</td> | ||
+ | <td width="70%">5‘-ACAACAGTTGCCGCTTGCATTTTAGCTTCCTTAGCTCCTG-3‘</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td width="30%" id="pSB1C3_Cm_del_rev">pSB1C3_Cm_del_rev (41 bp)</td> | ||
+ | <td width="70%">5‘-CGATGAAATACGTGGATTAATTTGATATCGAGCTCGCTTGG-3‘</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td width="30%" id="alr_Cm_del_fwd">alr_Cm_del_fwd (31 bp)</td> | ||
+ | <td width="70%">5‘-GGAAGCTAAAATGCAAGCGGCAACTGTTGTG-3‘</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td width="30%" id="dadX_Ec_control2">alr_Cm_del_rev (31 bp)</td> | ||
+ | <td width="70%">5‘-CGATATCAAATTAATCCACGTATTTCATCGC-3‘</td> | ||
+ | </tr> | ||
+ | <tr bgcolor="#D8D8D8"> | ||
+ | <th colspan="2" align="left">Konstitutive expression of <i>alr</i> for plasmid copy number for the antibiotic-free selection</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td width="30%" id="pSB_kon_pre_GFP">pSB_kon_pre_GFP (42 bp)</td> | ||
+ | <td width="70%">5‘-AGTTCTTCTCCTTTACGCATTTTCTCCTCTTTCTCTAGTAGC-3‘</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td width="30%" id="pSB_kon_suf_GFP">pSB_kon_suf_GFP (43 bp)</td> | ||
+ | <td width="70%">5‘-ACTACGCTTTAGTAGCTTAATACTAGTAGCGGCCGCTGCAGTC-3‘</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td width="30%" id="AB_kon_GFP_fwd">AB_kon_GFP_fwd (32 bp)</td> | ||
+ | <td width="70%">5‘-AAAGAGGAGAAAATGCGTAAAGGAGAAGAACT-3‘</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td width="30%" id="AB_kon_GFP_rev">AB_kon_GFP_rev (34 bp)</td> | ||
+ | <td width="70%">5‘-GCTACTAGTATTAAGCTACTAAAGCGTAGTTTTC-3‘</td> | ||
+ | </tr> | ||
+ | |||
+ | |||
</table> | </table> |
Latest revision as of 01:02, 18 October 2014
Primerlist
Primer ID | Primer sequence |
---|---|
General | |
VF-Primer (20 bp) | 5‘-CCACCTGACGTCTAAGAAAC-3‘ |
VR-Primer (22 bp) | 5‘-GTATTACCGCCTTTGAGTGAGC-3‘ |
Module 1 | |
Gene synthesis of the fumarate reductase (fumA, fumBCD) of B. succinogenes | |
pSB1C3_pre_Fum_A (43 bp) | 5‘-GTCTGCATTTTCTCCTCTTTCTCTAGAAGCGGCCGCGAATTCC-3‘ |
pSB1C3_suf_Fum_A (43 bp) | 5‘-CCAAAGCGCCGAATGCGTAATACTAGTAGCGGCCGCTGCAGTC-3‘ |
pSB1C3_pre_Fum_BCD (42 bp) | 5‘-TTGGTCATTTTCTCCTCTTTCTCTAGAAGCGGCCGCGAATCC-3‘ |
pSB1C3_suf_Fum_BCD (43 bp) | 5‘-GGGGTGTGGCGTCGATCTAATACTAGTAGCGGCCGCTGCAGTC-3‘ |
FumBCD_fwd (24 bp) | 5‘-CGGCCGCTTCTAGAGAAAGAGGAG-3‘ |
FumBCD_rev (22 bp) | 5‘-CGGCCGCTACTAGTATTAGATC-3‘ |
FumA_seq (20 bp) | 5‘-TGGAAGGCACCTTGGTTCAG-3‘ |
FumB_seq (20 bp) | 5‘-GCTCTGACCATGGCACATCG-3‘ |
Overexpression of the fumarate reductase (frd) of E. coli | |
T7_frd_Ec_fwd (64 bp) | 5‘-TAATACGACTCACTATAGGGAAAGAGGAGAAATACTAGATGCAAACCTTTCAAGCCGATCTTGC-3‘ |
T7_frd_Ec_rev (44 bp) | 5‘-TGCAGCGGCCGCTACTAGTATTAGATTGTAACGACACCAATCAG-3‘ |
pSB1C3_frd_pre (45 bp) | 5‘-GCAAGATCGGCTTGAAAGGTTTGCATCTAGTATTTCTCCTCTTTC-3‘ |
pSB1C3_frd_suf (47 bp) | 5‘-CTGATTGGTGTCGTTACAATCTAATACTAGTAGCGGCCGCTGCAGTC-3‘ |
frd_cut1_fwd (20 bp) | 5‘-CATACCGTTGCAGCAGAAGG-3‘ |
frd_cut1_rev (20 bp) | 5‘-CCTTCTGCTGCAACGGTATG-3‘ |
frd_cut2_fwd (20 bp) | 5‘-GACATGGAATTTGTTCAGTA-3‘ |
frd_cut2_rev (20 bp) | 5‘-TACTGAACAAATTCCATGTC-3‘ |
frd_cut3_fwd (20 bp) | 5‘-GAGCTGCAAGAACGCTTCAA-3‘ |
frd_cut3_rev (20 bp) | 5‘-TTGAAGCGTTCTTGCAGCTC-3‘ |
Replacement of the succinate carrier (dcuB) by porines (oprF) in the genome | |
porine_fwd_FRT (73 bp) | 5‘-TTATAAGAACCCGTACATCGCGGCGAAGATCCAGCCGAAGACGCACGATATTGACAGCTAGCTCAGTCCTAGG-3‘ |
porine_rev_FRT (46 bp) | 5‘-CGCCCTTTAGTGAGGGTTAATTTTACTTAGCTTGGGCTTCAACCT-3‘ |
pR6K_dcuB_fwd (46 bp) | 5‘-AGGTTGAAGCCCAAGCTAAGTAAAATTAACCCTCACTAAAGGGCG-3‘ |
pR6K_dcuB_rev (73 bp) | 5‘-ATGTTATTTACTATCCAACTTATCATAATACTGATATGTCTGTTTTATGGTAATACGACTCACTATAGGGCTC-3‘ |
dcuB_del_kon1 (20 bp) | 5‘-CGCCGCATTCTGTGAATAGC-3‘ |
dcuB_del_kon2 (20 bp) | 5‘-CCCTGGCACAAGTTACATCC-3‘ |
NADPH/NADP+ sensor (soxRS) | |
soxR_fwd (64 bp) | 5‘-TTGACAGCTAGCTCAGTCCTAGGTATTGTGCAAAGAGGAGAAAATGGAAAAGAAATTACCCCGC-3‘ |
soxR_rev (37 bp) | 5‘-GCTTCTAGAGTTAGTTTTGTTCATCTTCCAGCAAGCG-3‘ |
pSB1C3_suf_soxR (54 bp) | 5‘-TAGGACTGAGCTAGCTGTCAATACTAGTAGCTACTAGTAGCGGCCGCTGCAGTC-3‘ |
pSB1C3_pre_soxR (43 bp) | 5‘-TGGAAGATGAACAAAACTAACTCTAGAAGCGGCCGCGAATTCC-3‘ |
soxS_GFP_fwd (74 bp) | 5‘-TTATACTCCCCAACAGATGAATTAACGAACTGAACACTGAAAAGAGGCAGATTTATGCGTAAAGGAGAAGAACT-3‘ |
soxS_GFP_rev (42 bp) | 5‘-TGCAGCGGCCGCTACTAGTATATAAACGCAGAAAGGCCCACC-3‘ |
pSB1C3_pre_soxS (78 bp) | 5‘-TAATTCATCTGTTGGGGAGTATAATTCCTCAAGTTAACTTGAGGTAAAGCGATTTCTCTAGAAGCGGCCGCGAATTCC-3‘ |
pSB1C3_suf_soxS (43 bp) | 5‘-TGGGCCTTTCTGCGTTTATATACTAGTAGCGGCCGCTGCAGTC-3‘ |
Expression of the ccm cluster and cytochrome C (pccH) | |
ccm_fwd (58 bp) | 5‘-CTCAGTCCTAGGTATTGTGCAGAGAAAGAGGAGAAAATGCTTGAAGCCAGAGAGTTAC-3‘ |
ccm_rev (47 bp) | 5‘-TGCAGCGGCCGCTACTAGTATTATTTACTCTCCTGCGGCGACAAATG-3‘ |
pSB1C3_suf_ccm (50 bp) | 5‘-CATTTGTCGCCGCAGGAGAGTAAATAATACTAGTAGCGGCCGCTGCAGTC-3‘ |
pSB1C3_pre_ccm (28 bp) | 5‘-CTCTGCTAGCACAATACCTAGGACTGAG-3‘ |
pccH_fwd (55 bp) | 5‘-ATTCGCGGCCGCTTCTAGAGAAAGAGGAGAAAATGCAAAAAACAACTATTCTTTC-3‘ |
pccH_rev (40 bp) | 5‘-TGCAGCGGCCGCTACTAGTATCAGTACGTGACCGTGATGG-3‘ |
pSB1C3_pre_pccH (58 bp) | 5‘-GAAAGAATAGTTGTTTTTTGCATTTTCTCCTCTTTCTCTAGAAGCGGCCGCGAATTCC-3‘ |
pSB1C3_suf_pccH (43 bp) | 5‘-CCATCACGGTCACGTACTGATACTAGTAGCGGCCGCTGCAGTC-3‘ |
ccm_con1 (21 bp) | 5‘-ATTGGCATTGGCGGTTGGGTC-3‘ |
Module 2 | |
Gene synthesis of the RuBisCO of Synechoccus elangatus (rbcLS) and Halothiobacillus neapolitanus (ccbLS) and the phosphoribulokinase A (prkA) of Synechoccus elangatus | |
pSB1C3_pre_rbcLS (44 bp) | 5‘-CATCTAGTATTTCTCCTCTTTCTCTAGAAGCGGCCGCGAATTCC-3‘ |
pSB1C3_suf_rbcLS (43 bp) | 5‘-ATCGTCCGGGCCGCTACTAA-TACTAGTAGCGGCCGCTGCAGTC-3‘ |
pSB1C3_pre_ccbLS (44 bp) | 5‘-CATCTAGCGGCCGCTACTAGTATTAGATCTATTTCTCCTCTTTCTCTAGAAGCGGCCGCGAATTCC-3‘ |
pSB1C3_suf_ccbLS (43 bp) | 5‘-TGGTTTACCGCGGCAACTAATACTAGTAGCGGCCGCTGCAGTC-3‘ |
pSB1C3_pre_prkA (43 bp) | 5‘-ACACGATCTGGCTTGCTCATCTCTAGAAGCGGCCGCGAATTCC-3‘ |
pSB1C3_suf_prkA (43 bp) | 5‘-CGGTTGCCGCTAGCGTTTAATACTAGTAGCGGCCGCTGCAGTC-3‘ |
Hneap_seq (20 bp) | 5‘-TACAGTTTGGACCGACCTTC-3‘ |
Selan_seq (20 bp) | 5‘-GTACATGGACGACTGTATGG-3‘ |
Insert amplification of the gene synthesis of the RuBisCO of S. elangatus (SELAN) | |
Selan_fwd (20 bp) | 5‘-GGCCGCTTCTAGAGAAAGAG-3‘ |
Selan_rev (20 bp) | 5‘-GCGGCCGCTACTAGTATTAG-3‘ |
Sequencing primer for the addgene plasmid pHnCBcsoS1D | |
add_seq1 (20 bp) | 5‘-AGCGTGTCGTAGTGCGTATC-3‘ |
add_seq2 (20 bp) | 5‘-ACCACTTCTGGCTGCACAAC-3‘ |
add_seq3 (20 bp) | 5‘-CTGGCAACCGGCTTGATTAC-3‘ |
add_seq4 (20 bp) | 5‘-AAATCACCCACGCCAAACCG-3‘ |
add_seq5 (20 bp) | 5‘-GATGCGATTGGCTGCATACC-3‘ |
add_seq6 (20 bp) | 5‘-GCCGTAATATCCAGCTGAAC-3‘ |
Integration of the prkA in the plasdmid pHnCBcsoS1D | |
pHnCBcsoS1D_fwd (44 bp) | 5‘-CGGTTGCCGCTAGCGTTTAAGTGAGCGGATAACAATTTCAG-3‘ |
pHnCBcsoS1D_rev (45 bp) | 5‘-CATCTAGTATTTCTCCTCTTTTCCACACATTATACGAGCCGGAAG-3‘ |
prkA_pHn_rev (47 bp) | 5‘-CTGAAATTGTTATCCGCTCACTTAAACGCTAGCGGCAACCGGTGCTG-3‘ |
prkA_pHn_fwd (53 bp) | 5‘-CTTCCGGCTCGTATAATGTGTGGAAAAGAGGAGAAATACTAGATGAGCAAGCC-3‘ |
Shell associated protein of the carboxysome (sap respectively csoS2) | |
csoS2_sap_1_fwd (43) | 5‘-ATTCGCGGCCGCTTCTAGAGGTCAAGTCTGTCATTGCGCTGTG-3‘ |
pSB1C3_pre_sap_1 (43 bp) | 5‘-AGCGCAATGACAGACTTGACCTCTAGAAGCGGCCGCGAATTCC-3‘ |
csoS2_sap_1_rev (38 bp) | 5‘-GCGCCGAACATGACTTTATCCGCTTGTGAAGGCTTTGC-3‘ |
sap_2_fwd (22 bp) | 5‘-AAGTCATGTTCGGCGCCACAGC-3‘ |
sap_2_rev (20 bp) | 5‘-GCCGCTACTAGTATTAACCG-3‘ |
pSB1C3_suf_sap2 (40 bp) | 5‘-CCGGCGGCGCGCGCGGTTAATACTAGTAGCGGCCGCTGCA-3‘ |
seq_sap_2 (20 bp) | 5‘-TGCCCGTCACCGGTAATGAG-3‘ |
Carbonic anhydrase of the carboxysome (can) | |
csoS3_can_fwd (50 bp) | 5‘-ATTCGCGGCCGCTTCTAGAGTTAAGTAAAGTGTAACGATCATGAACACCC-3‘ |
pSB1C3_pre_can (46 bp) | 5‘-GTTCATGATCGTTACACTTTACTTAACTCTAGAAGCGGCCGCGAAT-3‘ |
csoS3_can_rev (41 bp) | 5‘-TGCAGCGGCCGCTACTAGTATCATGCGGATGCAACCTCTTC-3‘ |
pSB1C3_suf_can (46 bp) | 5‘-TCGAAGAGGTTGCATCCGCATGATACTAGTAGCGGCCGCTGCAGTC-3‘ |
Shell proteins of the carboxysome (csoS4AB and csoS1CAB) | |
fw_pSB1C3_csoS4A (47 bp) | 5‘-ATTCGCGGCCGCTTCTAGAGAAATCATGCAAGTTGAGAAAACG-3‘ |
rv_csoS4A_PstI (43 bp) | 5‘-GAGGGATAAGACTTGCTTCCGGCAGCCTCTCGTGCTGCCGATG-3‘ |
fw_PstI_csoS4A (51 bp) | 5‘-TCATCGGCAGCACGAGAGGCTGCCGGAAGCAAGTCTTATCCCTCTGATTTG-3‘ |
rv_SpeI_Intergen (52 bp) | 5‘-GTCGTTGCCATAACAGATCCTCTAAAACTAGGTTGAGCACGTTCAGAACGAG-3‘ |
fw_SpeI_Intergen (49 bp) | 5‘-TCGTTCTGAACGTGCTCAACCTAGTTTTAGAGGATCTGTTATGGCAACG-3‘ |
rv_csoS1_pSB1C3 (50 bp) | 5‘-CGGACTGCAGCGGCCGCTACTAGTATTAGCTATTCAGATTTGCGATACAC-3‘ |
fw_csoS1_pSB1C3 (46 bp) | 5‘-GTGTATCGCAAATCTGAATAGCTAATACTAGTAGCGGCCGCTGCAG-3‘ |
rv_pSB1C3_csoS4A (47 bp) | 5‘-AACGTTTTCTCAACTTGCATGATTTCTCTAGAAGCGGCCGCGAATTC-3‘ |
Pore protein of the carboxysome (csoS1D) | |
csoS1D_fwd (40 bp) | 5‘-ATTCGCGGCCGCTTCTAGAGTGTCTAGTAGGGAAGATGCG-3‘ |
csoS1D_rev (41 bp) | 5‘-TGCAGCGGCCGCTACTAGTATTAGAACCCTTCAGCGCGACG-3‘ |
pSB1C3_suf_csoS1D (43bp) | 5‘-GTCGCGCTGAAGGGTTCTAATACTAGTAGCGGCCGCTGCAGTC-3‘ |
pSB1C3_pre_csoS1D (43 bp) | 5‘-CGCATCTTCCCTACTAGACACTCTAGAAGCGGCCGCGAATTCC-3‘ |
GFP fusion with the shell proteins and the shell-associated protein | |
fw-csoS1A-GFP (45 bp) | 5‘-CTGCCTAAGGCGCCACAAGCCATGCGTAAAGGAGAAGAACTTTTC-3‘ |
rv-csoS1B-GFP (55 bp) | 5‘-GTGAGCCGTCTTAGGAATATCTGACTTATTATTTGTATAGTTCATCCATGCCATG-3‘ |
fw-GFP-csoS1B (48 bp) | 5‘-CATGGATGAACTATACAAATAATAAGTCAGATATTCCTAAGACGGCTC-3‘ |
rv-GFP-csoS1A (45 bp) | 5‘-GAAAAGTTCTTCTCCTTTACGCATGGCTTGTGGCGCCTTAGGCAG-3‘ |
Gluco switch using sRNA | |
pSB1C3_pre_sRNApfkA (43 bp) | 5‘-TGTGGCTAAAAGAATTAGTACTCTAGAAGCGGCCGCGAATTCC-3‘ |
pSB1C3_suf_sRNApfkA (43 bp) | 5‘-TGGGCCTTTCTGCGTTTATATACTAGTAGCGGCCGCTGCAGTC-3‘ |
sRNA:pfkB_fwd (46 bp) | 5‘-TACCATGCATAGATATGCAACTGTTTTCTGTTGGGCCATTGCATTG-3‘ |
sRNA:pfkB_rev (47 bp) | 5‘-ACAGTTGCATATCTATGCATGGTACTCTAGAAGCGGCCGCGAATTCC-3‘ |
Amplification of purification vector with T7, RBS and intein tag with chitin binding domain | |
int_RBS_fw (42 bp) | 5‘-CTATAGGGAAAGAGGAGAAATGCATCACGGGAGATGCGCTAG-3‘ |
RBS_int_rev (44 bp) | 5‘-CTAGCGCATCTCCCGTGATGCATTTCTCCTCTTTCCCTATAGTG-3‘ |
RBS_rev (22 bp) | 5‘-TTTCTCCTCTTTCCCTATAGTG-3‘ |
int_fw (22 bp) | 5‘-TGCATCACGGGAGATGCACTAG-3‘ |
int_pSB1C3_rev (50 bp) | 5‘-GACTGCAGCGGCCGCTACTAGTATCATTGAAGCTGCCACAAGGCAGGAAC-3‘ |
pSB1C3_int_fw (46 bp) | 5‘-CTGCCTTGTGGCAGCTTCAATGATACTAGTAGCGGCCGCTGCAGTC-3‘ |
Expression of the SBPase (glpX) for the calvin cycle and for purification | |
fw-pSB1C3-BBa_B0034-SBPase (58 bp) | 5‘-GAATTCGCGGCCGCTTCTAGAGAAAGAGGAGAAAATGAGGGAATTGAAAAGCGAAAAG-3‘ |
rv-pSB1C3-SBPase (56 bp) | 5‘-GACTGCAGCGGCCGCTACTAGTATCAATGATAAGCTTCAATAAATTGGTATTTTAC-3‘ |
fw-SBPase-Schnittstelle (56 bp) | 5‘-AAAAGCAAAAGGCTGCGTAAATTTGGACGCATCTTTAACAGAAAATATGAAATCAG-3‘ |
rv-SBPase-Schnittstelle (46 bp) | 5‘-TTTTCTGTTAAAGATGCGTCCAAATTTACGCAGCCTTTTGCTTTTG-3‘ |
fw-SBPase-pSB1C3 (57 bp) | 5‘-AATTTATTGAAGCTTATCATTGATACTAGTAGCGGCCGCTGCAGTCCGGCAAAAAAG-3‘ |
rv-SBPase-BBa_B0034-pSB1C3 (65 bp) | 5‘-TTTTCGCTTTTCAATTCCCTCATTTTCTCCTCTTTCTCTAGAAGCGGCCGCGAATTCCAGAAATC-3‘ |
SBPase_purify_fw (45 bp) | 5‘-CTATAGGGAAAGAGGAGAAATATGAGGGAATTGAAAAGCGAAAAG-3‘ |
SBPase_purify_rev (53 bp) | 5‘-CTAGTGCATCTCCCGTGATGCACGTAAAATACCAATTTATTGAAGCTTATCAT-3‘ |
fw_pSB1_SBPase (58 bp) | 5‘-GAATTCGCGGCCGCTTCTAGAGAAAGAGGAGAAAATGAGGGAATTGAAAAGCGAAAAG-3‘ |
rv_pSB1_SBPase (56 bp) | 5‘-GACTGCAGCGGCCGCTACTAGTATCAATGATAAGCTTCAATAAATTGGTATTTTAC-3‘ |
fw_SBPase_XbaI (56 bp) | 5‘-AAAAGCAAAAGGCTGCGTAAATTTGGACGCATCTTTAACAGAAAATATGAAATCAG-3‘ |
rv_SBPase_XbaI (46 bp) | 5‘-TTTTCTGTTAAAGATGCGTCCAAATTTACGCAGCCTTTTGCTTTTG-3‘ |
fw_SBPase_pSB1C3 (57 bp) | 5‘-AATTTATTGAAGCTTATCATTGATACTAGTAGCGGCCGCTGCAGTCCGGCAAAAAAG-3‘ |
rv_SBPase_pSB1 (65 bp) | 5‘-TTTTCGCTTTTCAATTCCCTCATTTTCTCCTCTTTCTCTAGAAGCGGCCGCGAATTCCAGAAATC-3‘ |
Amplification of E. coli fructose bisphosphate aldolase II (fbaA) for purification | |
fbaA_purify_fw (50 bp) | 5‘-CTATAGGGAAAGAGGAGAAATATGTCTAAGATTTTTGATTTCGTAAAACC-3‘ |
fbaA_purify_rev (41 bp) | 5‘-CTAGTGCATCTCCCGTGATGCACAGAACGTCGATCGCGTTC-3‘ |
fbaA_PstI_fw (42 bp) | 5‘-CAAAGCGAACGAAGCTTATCTGCAAGGTCAGCTGGGTAACCC-3‘ |
fbaA_PstI_rev (43 bp) | 5‘-CTTTCGGGTTACCCAGCTGACCTTGCAGATAAGCTTCGTTCGC-3‘ |
Amplification of E. coli transketolase I (tktA) for purification | |
tktA_purify_fw (43 bp) | 5‘-CTATAGGGAAAGAGGAGAAATATGTCCTCACGTAAAGAGCTTG-3‘ |
tktA_purify_rev (43 bp) | 5‘-CTAGTGCATCTCCCGTGATGCACAGCAGTTCTTTTGCTTTCGC-3‘ |
tktA_PstI_fw (41 bp) | 5‘-GTTCGGTCCGCTGTTGCCGGAACTCCTCGGCGGTTCTGCTG-3‘ |
tktA_PstI_rev (42 bp) | 5‘-CAGGTCAGCAGAACCGCCGAGGAGTTCCGGCAACAGCGGACC-3‘ |
Amplification of C. glutamicum phosphoglycerate kinase (pgk) for purification | |
cg_pgk_purify_fw (42 bp) | 5‘-CTATAGGGAAAGAGGAGAAATATGGCTGTTAAGACCCTCAAG-3‘ |
cg_pgk_purify_rev (42 bp) | 5‘-CTAGTGCATCTCCCGTGATGCACTGAGCGAGAATTGCAACGC-3‘ |
Amplification of E. coli glyceraldehyde 3-phosphate dehydrogenase (gapA) for purification | |
gapA_purify_fw (47 bp) | 5‘-CTATAGGGAAAGAGGAGAAATATGACTATCAAAGTAGGTATCAACGG-3‘ |
gapA_purify_rev (43 bp) | 5‘-CTAGTGCATCTCCCGTGATGCATTTGGAGATGTGAGCGATCAG-3‘ |
gapA_NotI_fw (44 bp) | 5‘-CTCACAAAGACTGGCGCGGCGGCCGTGGCGCTTCCCAGAACATC-3‘ |
gapA_NotI_rev (43 bp) | 5‘-GATGATGTTCTGGGAAGCGCCACGGCCGCCGCGCCAGTCTTTG-3‘ |
Amplification of S. elongatus phosphoribulokinase (prkA) for purification | |
prkA_purify_fw (40 bp) | 5‘-CTATAGGGAAAGAGGAGAAATATGAGCAAGCCAGATCGTG-3‘ |
prkA_purify_rev (42 bp) | 5‘-CTAGTGCATCTCCCGTGATGCAAACGCTAGCGGCAACCGGTG-3‘ |
Amplification of E. coli pyruvate kinase (pykF) for purification | |
pykF_purify_fw (47 bp) | 5‘-CTATAGGGAAAGAGGAGAAATATGAAAAAGACCAAAATTGTTTGCAC-3‘ |
pykF_purify_rev (42 bp) | 5‘-CTAGTGCATCTCCCGTGATGCACAGGACGTGAACAGATGCGG-3‘ |
pykF_PstI_fw (48 bp) | 5‘-ACCGTCTGGGTAAAGAACTGGCTCTGCAAAGCGGTCTGGCACACAAAG-3‘ |
pykF_PstI_rev (43 bp) | 5‘-CACCTTTGTGTGCCAGACCGCTTTGCAGAGCCAGTTCTTTACC-3‘ |
Amplification of E. coli D-lactate dehydrogenase (ldhA) for purification | |
ldhA_purify_fw (44 bp) | 5‘-CTATAGGGAAAGAGGAGAAATATGAAACTCGCCGTTTATAGCAC-3‘ |
ldhA_purify_rev (42 bp) | 5‘-CTAGTGCATCTCCCGTGATGCAAACCAGTTCGTTCGGGCAGG-3‘ |
Amplification of E. coli glucokinase (glk) for purification | |
glk_purify_fw (44 bp) | 5‘-CTATAGGGAAAGAGGAGAAATATGACAAAGTATGCATTAGTCGG-3‘ |
glk_purify_rev (43 bp) | 5‘-CTAGTGCATCTCCCGTGATGCACAGAATGTGACCTAAGGTCTG-3‘ |
Amplification of E. coli glucose 6-phosphate-1-dehydrogenase (zwf) for purification | |
zwf_purify_fw (41 bp) | 5‘-CTATAGGGAAAGAGGAGAAATATGGCGGTAACGCAAACAGC-3‘ |
zwf_purify_rev (45 bp) | 5‘-CTAGTGCATCTCCCGTGATGCACTCAAACTCATTCCAGGAACGAC-3‘ |
Module 3 | |
Backbone for the isobutanol plasmid | |
fw_kivD_pSB1C3 (53 bp) | 5‘-GCAAACTATTTGCTGAACAAAATAAATCATAATACTAGTAGCGGCCGCTGCAG-3‘ |
rv_alsS_pSB1C3 (65 bp) | 5‘-TTTTTGTTCTTTTGTTGCTTTTGTCAACATTTTCTCCTCTTTCTCTAGAAGCGGCCGCGAATTC-3‘ |
fw_adhA_pSB1C3 (51 bp) | 5‘-GGCCGAATGGTCATTGATTTTACTAAATAATACTAGTAGCGGCCGCTGCAG-3‘ |
fw_adhA_pSB1C3 (42 bp) | 5‘-CATTGATTTTACTAAATAATACTAGTAGCGGCCGCTGCAGTC-3‘ |
alsS | |
fw_pSB1C3_alsS (79 bp) | 5‘-TTCGCTAAGGATGATTTCTGGAATTCGCGGCCGCTTCTAGAGAAAGAGGAGAAAATGTTGACAAAAGCAACAAAAGAAC-3‘ |
rv_ilvC_alsS (64 bp) | 5‘-GATTCAGTGTATTGAAGTAGTTAGCCATTTTCTCCTCTTTCTAGAGAGCTTTCGTTTTCATGAG-3‘ |
ilvC | |
fw_alsS_ilvC (65 bp) | 5‘-CTCATGAAAACGAAAGCTCTCTAGAAAGAGGAGAAAATGGCTAACTACTTCAATACACTGAATC-3‘ |
rev_ilvD_ilvC (56 bp) | 5‘-GTGGCGGAACGGTACTTAGGCATTTTCTCCTCTTTTTAACCCGCAACAGCAATACG-3‘ |
ilvD | |
fw_ilvC_ilvD (57 bp) | 5‘-CGTATTGCTGTTGCGGGTTAAAAAGAGGAGAAAATGCCTAAGTACCGTTCCGCCAC-3‘ |
rv_kivD_ilvD (67 bp) | 5‘-GTCTAATAGGTAATCTCCTACTGTATACATTTTCTCCTCTTTTTAACCCCCCAGTTTCGATTTATC-3‘ |
kivD | |
fw_ilvD_kivD (67 bp) | 5‘-GATAAATCGAAACTGGGGGGTTAAAAAGAGGAGAAAATGTATACAGTAGGAGATTACCTATTAGAC-3‘ |
rv_pSB1C3_kivD (74 bp) | 5‘-ACCTTGCCCTTTTTTGCCGGACTGCAGCGGCCGCTACTAGTATTATGATTTATTTTGTTCAGCAAATAGTTTGC-3‘ |
rv_adhA_kivD (69 bp) | 5‘-GTGTCTTACTACTGCTGCTTTCATTTTCTCCTCTTTTTATGATTTATTTTGTTCAGCAAATAGTTTGC-3‘ |
Alcohol dehydrogenase (adhA) of Lactococcus lactis | |
fw_kivD_adhA (69 bp) | 5‘-GCAAACTATTTGCTGAACAAAATAAATCATAAAAAGAGGAGAAAATGAAAGCAGCAGTAGTAAGACAC-3‘ |
rev_pSB1C3_adhA (80 bp) | 5‘-GTGACACCTTGCCCTTTTTTGCCGGACTGCAGCGGCCGCTACTAGTATTATTTAGTAAAATCAATGACCATTCGGCCTTC-3‘ |
fw_pSB1C3_adhA (66 bp) | 5‘-GATTTCTGGAATTCGCGGCCGCTTCTAGAGAAAGAGGAGAAAATGAAAGCAGCAGTAGTAAGACAC-3‘ |
rev_adhA_pSB1C3 (56 bp) | 5‘-GTCTTACTACTGCTGCTTTCATTTTCTCCTCTTTCTCTAGAAGCGGCCGCGAATTC-3‘ |
fw-pSB1C3-BBa_B0034-adhaA (66 bp) | 5‘-GATTTCTGGAATTCGCGGCCGCTTCTAGAGAAAGAGGAGAAAATGAAAGCAGCAGTAGTAAGACAC-3‘ |
rev-adhA-BBa_B0034-pSB1C3-adhaA (56 bp) | 5‘-GTCTTACTACTGCTGCTTTCATTTTCTCCTCTTTCTCTAGAAGCGGCCGCGAATTC-3‘ |
Illegal restriction site between alsS and ilvC | |
rv_ilvC_alsS-new (72 bp) | 5‘-CAGTGTATTGAAGTAGTTAGCCATTTTCTCCTCTTTTTAGACAGTTTCCGTTTTCATGAGTTCCCCAAATTC-3‘ |
fw_alsS_ilvC-new (62 bp) | 5‘-CATGAAAACGGAAACTGTCTAAAAAGAGGAGAAAATGGCTAACTACTTCAATACACTGAATC-3‘ |
Biosafety and antibiotic-free selection | |
Deletion of alanine racemases (alr and dadX) | |
alr_fwd_redET (72 bp) | 5‘-AATTAGGTAATTAAAGCAAACACTTATCAAGGAACACAAATGCAAGCGGCAATTAACCCTCACTAAAGGGCG-3‘ |
alr_rev_redEt (73 bp) | 5‘-TTTTATTGGCCGGATAAGGCATTCACGCCGCATCCGGCACAGACAATCAATAATACGACTCACTATAGGGCTC-3‘ |
dadX_fwd_redET (72 bp) | 5‘-CGTCCGGGCCATTTACATGGCGCACACAGCTAAGGAAACGAGATGACCCAAATTAACCCTCACTAAAGGGCG-3‘ |
dadX_rev_redET (73 bp) | 5‘-TGATTTTTTTGCACCCAGAAGACGTTGCCTCCGATCCGGCTTACAACAAGTAATACGACTCACTATAGGGCTC-3‘ |
alr_Ec_control1 (20 bp) | 5‘-GCTGGAGATGCCATCAGAAC-3‘ |
alr_Ec_control2 (20 bp) | 5‘-CCGGCGAATATTGCTACGTG-3‘ |
dadX_Ec_control1 (20 bp) | 5‘-GCTTTAATACGCCCGTTGAC-3‘ |
dadX_Ec_control2 (20 bp) | 5‘-CTGGATCAACGCTTCTTTGG-3‘ |
Replacement of the chloramphenicol resistance (Cm) by the konstitutive alanine racemase alr | |
pSB1C3_Cm_del_fwd (40 bp) | 5‘-ACAACAGTTGCCGCTTGCATTTTAGCTTCCTTAGCTCCTG-3‘ |
pSB1C3_Cm_del_rev (41 bp) | 5‘-CGATGAAATACGTGGATTAATTTGATATCGAGCTCGCTTGG-3‘ |
alr_Cm_del_fwd (31 bp) | 5‘-GGAAGCTAAAATGCAAGCGGCAACTGTTGTG-3‘ |
alr_Cm_del_rev (31 bp) | 5‘-CGATATCAAATTAATCCACGTATTTCATCGC-3‘ |
Konstitutive expression of alr for plasmid copy number for the antibiotic-free selection | |
pSB_kon_pre_GFP (42 bp) | 5‘-AGTTCTTCTCCTTTACGCATTTTCTCCTCTTTCTCTAGTAGC-3‘ |
pSB_kon_suf_GFP (43 bp) | 5‘-ACTACGCTTTAGTAGCTTAATACTAGTAGCGGCCGCTGCAGTC-3‘ |
AB_kon_GFP_fwd (32 bp) | 5‘-AAAGAGGAGAAAATGCGTAAAGGAGAAGAACT-3‘ |
AB_kon_GFP_rev (34 bp) | 5‘-GCTACTAGTATTAAGCTACTAAAGCGTAGTTTTC-3‘ |