Team:Bielefeld-CeBiTec/Notebook/Primer

From 2014.igem.org

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<table width="100%" cellspacing="10" align="center" style="background-color:transparent">
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  <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>
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</tr>
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</table>
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</div>
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<br>
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</tr>
</tr>
<tr bgcolor="#3fb498">
<tr bgcolor="#3fb498">
-
   <th colspan="2" align="left">?????????</th>
+
   <th colspan="2" align="left" id="General">General</th>
</tr>
</tr>
<tr>
<tr>
-
<td width="30%">sRNA:pfkB_fwd (46 bp)</td>
+
<td width="30%" id="VF-Primer">VF-Primer (20 bp)</td>
-
<td width="70%">5‘-TACCATGCATAGATATGCAACTGTTTTCTGTTGGGCCATTGCATTG-3‘</td>
+
-
</tr>
+
-
<tr bgcolor="#3fb498">
+
-
  <th colspan="2" align="left">General</th>
+
-
</tr>
+
-
<tr>
+
-
<td width="30%">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">CCM-Cassette (<i>E. coli</i>)</th>
+
   <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 bgcolor="#D8D8D8">
 
-
  <th colspan="2" align="left">Cytochrom C (<i>Geobacter sulfurreducens</i>)</th>
 
</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">??? (Julian)</th>
+
   <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%">T7_frd_Ec_fwd (64 bp)</td>
+
<td width="30%" id="pSB1C3_pre_rbcLS">pSB1C3_pre_rbcLS (44 bp)</td>
-
<td width="70%">5‘-TAATACGACTCACTATAGGGAAAGAGGAGAAATACTAGATGCAAACCTTTCAAGCCGATCTTGC-3‘</td>
+
<td width="70%">5‘-CATCTAGTATTTCTCCTCTTTCTCTAGAAGCGGCCGCGAATTCC-3‘</td>
</tr>
</tr>
<tr>
<tr>
-
<td width="30%">T7_frd_Ec_rev (44 bp)</td>
+
<td width="30%" id="pSB1C3_suf_rbcLS">pSB1C3_suf_rbcLS (43 bp)</td>
-
<td width="70%">5‘-TGCAGCGGCCGCTACTAGTATTAGATTGTAACGACACCAATCAG-3‘</td>
+
<td width="70%">5‘-ATCGTCCGGGCCGCTACTAA-TACTAGTAGCGGCCGCTGCAGTC-3‘</td>
</tr>
</tr>
<tr>
<tr>
-
<td width="30%">pSB1C3_frd_pre (45 bp)</td>
+
<td width="30%" id="pSB1C3_pre_ccbLS">pSB1C3_pre_ccbLS (44 bp)</td>
-
<td width="70%">5‘-GCAAGATCGGCTTGAAAGGTTTGCATCTAGTATTTCTCCTCTTTC-3‘</td>
+
<td width="70%">5‘-CATCTAGCGGCCGCTACTAGTATTAGATCTATTTCTCCTCTTTCTCTAGAAGCGGCCGCGAATTCC-3‘</td>
</tr>
</tr>
<tr>
<tr>
-
<td width="30%">pSB1C3_frd_suf (47 bp)</td>
+
<td width="30%" id="pSB1C3_suf_ccbLS">pSB1C3_suf_ccbLS (43 bp)</td>
-
<td width="70%">5‘-CTGATTGGTGTCGTTACAATCTAATACTAGTAGCGGCCGCTGCAGTC-3‘</td>
+
<td width="70%">5‘-TGGTTTACCGCGGCAACTAATACTAGTAGCGGCCGCTGCAGTC-3‘</td>
</tr>
</tr>
<tr>
<tr>
-
<td width="30%">pR6K_dcuB_short (22 bp)</td>
+
<td width="30%" id="pSB1C3_pre_prkA">pSB1C3_pre_prkA (43 bp)</td>
-
<td width="70%">5‘-AATTAACCCTCACTAAAGGGCG-3‘</td>
+
<td width="70%">5‘-ACACGATCTGGCTTGCTCATCTCTAGAAGCGGCCGCGAATTCC-3‘</td>
</tr>
</tr>
-
<tr bgcolor="#3fb498">
+
<tr>
-
  <th colspan="2" align="left">Module 2</th>
+
<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">Carbonic anhydrase</th>
+
   <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%">csoS3_can_fwd (50 bp)</td>
+
<td width="30%" id="Selan_fwd">Selan_fwd (20 bp)</td>
-
<td width="70%">5‘-ATTCGCGGCCGCTTCTAGAGTTAAGTAAAGTGTAACGATCATGAACACCC-3‘</td>
+
<td width="70%">5‘-GGCCGCTTCTAGAGAAAGAG-3‘</td>
</tr>
</tr>
<tr>
<tr>
-
<td width="30%">pSB1C3_pre_can (46 bp)</td>
+
<td width="30%" id="Selan_rev">Selan_rev (20 bp)</td>
-
<td width="70%">5‘-GTTCATGATCGTTACACTTTACTTAACTCTAGAAGCGGCCGCGAAT-3‘</td>
+
<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%">csoS3_can_rev (41 bp)</td>
+
<td width="30%" id="add_seq1">add_seq1 (20 bp)</td>
-
<td width="70%">5‘-TGCAGCGGCCGCTACTAGTATCATGCGGATGCAACCTCTTC-3‘</td>
+
<td width="70%">5‘-AGCGTGTCGTAGTGCGTATC-3‘</td>
</tr>
</tr>
<tr>
<tr>
-
<td width="30%">pSB1C3_suf_can (46 bp)</td>
+
<td width="30%" id="add_seq2">add_seq2 (20 bp)</td>
-
<td width="70%">5‘-TCGAAGAGGTTGCATCCGCATGATACTAGTAGCGGCCGCTGCAGTC-3‘</td>
+
<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">Shell associated protein</th>
+
   <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">csoS1D</th>
+
   <th colspan="2" align="left">Carbonic anhydrase of the carboxysome (<i>can</i>)</th>
</tr>
</tr>
<tr>
<tr>
-
<td width="30%">csoS1D_fwd (40 bp)</td>
+
<td width="30%" id="csoS3_can_fwd">csoS3_can_fwd (50 bp)</td>
-
<td width="70%">5‘-ATTCGCGGCCGCTTCTAGAGTGTCTAGTAGGGAAGATGCG-3‘</td>
+
<td width="70%">5‘-ATTCGCGGCCGCTTCTAGAGTTAAGTAAAGTGTAACGATCATGAACACCC-3‘</td>
</tr>
</tr>
<tr>
<tr>
-
<td width="30%">csoS1D_rev (41 bp)</td>
+
<td width="30%" id="pSB1C3_pre_can">pSB1C3_pre_can (46 bp)</td>
-
<td width="70%">5‘-TGCAGCGGCCGCTACTAGTATTAGAACCCTTCAGCGCGACG-3‘</td>
+
<td width="70%">5‘-GTTCATGATCGTTACACTTTACTTAACTCTAGAAGCGGCCGCGAAT-3‘</td>
</tr>
</tr>
<tr>
<tr>
-
<td width="30%">pSB1C3_suf_csoS1D (43bp)</td>
+
<td width="30%" id="csoS3_can_rev">csoS3_can_rev (41 bp)</td>
-
<td width="70%">5‘-GTCGCGCTGAAGGGTTCTAATACTAGTAGCGGCCGCTGCAGTC-3‘</td>
+
<td width="70%">5‘-TGCAGCGGCCGCTACTAGTATCATGCGGATGCAACCTCTTC-3‘</td>
</tr>
</tr>
<tr>
<tr>
-
<td width="30%">pSB1C3_pre_csoS1D (43 bp)</td>
+
<td width="30%" id="pSB1C3_suf_can">pSB1C3_suf_can (46 bp)</td>
-
<td width="70%">5‘-CGCATCTTCCCTACTAGACACTCTAGAAGCGGCCGCGAATTCC-3‘</td>
+
<td width="70%">5‘-TCGAAGAGGTTGCATCCGCATGATACTAGTAGCGGCCGCTGCAGTC-3‘</td>
</tr>
</tr>
<tr bgcolor="#D8D8D8">
<tr bgcolor="#D8D8D8">
-
   <th colspan="2" align="left">Second part</th>
+
   <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%">rv-csoS1-pSB1C3 (50 bp)</td>
+
<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%">fw-csoS1-pSB1C3 (46 bp)</td>
+
<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%">rv-pSB1C3-csoS4A (47 bp)</td>
+
<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">Gluco switch</th>
+
   <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">General primer</th>
+
   <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 (1) (42 bp)</td>
+
<td width="30%" id="int_RBS_fw">int_RBS_fw (42 bp)</td>
-
<td width="70%">5‘-CTATAGGGAAAGAGGAGAAATGCATCACGGGAGATGCACTAG-3‘</td>
+
<td width="70%">5‘-CTATAGGGAAAGAGGAGAAATGCATCACGGGAGATGCGCTAG-3‘</td>
</tr>
</tr>
<tr>
<tr>
-
<td width="30%">RBS_int_rev (2) (44 bp)</td>
+
<td width="30%" id="RBS_int_rev">RBS_int_rev (44 bp)</td>
-
<td width="70%">5‘-CTAGTGCATCTCCCGTGATGCATTTCTCCTCTTTCCCTATAGTG-3‘</td>
+
<td width="70%">5‘-CTAGCGCATCTCCCGTGATGCATTTCTCCTCTTTCCCTATAGTG-3‘</td>
</tr>
</tr>
<tr>
<tr>
-
<td width="30%">RBS_rev (3) (22 bp)</td>
+
<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 (4) (22 bp)</td>
+
<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 (5) (50 bp)</td>
+
<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 (6) (46 bp)</td>
+
<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">cg_pgk</th>
+
   <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 bgcolor="#D8D8D8">
 
-
  <th colspan="2" align="left">Gene synthesis</th>
 
-
</tr>
 
-
<tr>
 
-
<td width="30%">pSB1C3_pre_rbcLS (44 bp)</td>
 
-
<td width="70%">5‘-CATCTAGTATTTCTCCTCTTTCTCTAGAAGCGGCCGCGAATTCC-3‘</td>
 
-
</tr>
 
-
<tr>
 
-
<td width="30%">pSB1C3_suf_rbcLS (43 bp)</td>
 
-
<td width="70%">5‘-ATCGTCCGGGCCGCTACTAA-TACTAGTAGCGGCCGCTGCAGTC-3‘</td>
 
-
</tr>
 
-
<tr>
 
-
<td width="30%">pSB1C3_pre_ccbLS (44 bp)</td>
 
-
<td width="70%">5‘-CATCTAGTATTTCTCCTCTTTCTCTAGAAGCGGCCGCGAATTCC-3‘</td>
 
-
</tr>
 
-
<tr>
 
-
<td width="30%">pSB1C3_suf_ccbLS (43 bp)</td>
 
-
<td width="70%">5‘-TGGTTTACCGCGGCAACTAATACTAGTAGCGGCCGCTGCAGTC-3‘</td>
 
-
</tr>
 
-
<tr>
 
-
<td width="30%">pSB1C3_pre_prkA (43 bp)</td>
 
-
<td width="70%">5‘-ACACGATCTGGCTTGCTCATCTCTAGAAGCGGCCGCGAATTCC-3‘</td>
 
-
</tr>
 
-
<tr>
 
-
<td width="30%">pSB1C3_suf_prkA (43 bp)</td>
 
-
<td width="70%">5‘-CGGTTGCCGCTAGCGTTTAATACTAGTAGCGGCCGCTGCAGTC-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%">fw-kivD-pSB1C3 (53 bp)</td>
+
<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%">rv-alsS-pSB1C3 (65 bp)</td>
+
<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%">fw-adhA-pSB1C3 (43 bp)</td>
+
<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%">fw-pSB1C3-alsS (79 bp)</td>
+
<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%">rv-ilvC-alsS (64 bp)</td>
+
<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%">fw-alsS-ilvC (65 bp)</td>
+
<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%">rev-ilvD-ilvC (56 bp)</td>
+
<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%">fw-ilvC-ilvD (57 bp)</td>
+
<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%">rv-kivD-ilvD (67 bp)</td>
+
<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%">fw-ilvD-kivD (67 bp)</td>
+
<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%">rv-pSB1C3-kivD (74 bp)</td>
+
<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%">rv-adhA-kivD (69 bp)</td>
+
<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%">fw-kivD-adhA (69 bp)</td>
+
<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%">rev-pSB1C3-adhA (80 bp)</td>
+
<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


General

Module 1

Module 2

Module 3

Biosafety

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‘

Retrieved from "http://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/Primer"