Team:Bielefeld-CeBiTec/Notebook/Primer

From 2014.igem.org

(Difference between revisions)
Line 42: Line 42:
   <th width="30%" align="center">Primer ID</th>
   <th width="30%" align="center">Primer ID</th>
   <th width="70%" align="center">Primer sequence</th>
   <th width="70%" align="center">Primer sequence</th>
-
</tr>
 
-
<tr bgcolor="#3fb498">
 
-
  <th colspan="2" align="left">?????????</th>
 
-
</tr>
 
-
<tr>
 
-
<td width="30%" id="sRNA:pfkb_fwd">sRNA:pfkB_fwd (46 bp)</td>
 
-
<td width="70%">5‘-TACCATGCATAGATATGCAACTGTTTTCTGTTGGGCCATTGCATTG-3‘</td>
 
</tr>
</tr>
<tr bgcolor="#3fb498">
<tr bgcolor="#3fb498">
Line 65: Line 58:
</tr>
</tr>
<tr bgcolor="#D8D8D8">
<tr bgcolor="#D8D8D8">
-
   <th colspan="2" align="left">Gene synthesis of the fumarate reductase of <i>B. succinogenes</i></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>
Line 100: Line 93:
</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">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>
 +
<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>
 +
<td width="30%" id="pR6K_dcuB_short">pR6K_dcuB_short (22 bp) </td>
 +
<td width="70%">5‘-AATTAACCCTCACTAAAGGGCG-3‘</td>
 +
</tr>
 +
<tr bgcolor="#D8D8D8">
 +
  <th colspan="2" align="left">NAPH/NAP+ 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 bgcolor="#D8D8D8">
 +
  <th colspan="2" align="left">Expression of the ccm cluster and cytochrome C (<i>pccH</i>)</th>
</tr>
</tr>
<tr>
<tr>
Line 117: Line 196:
<td width="30%"id="pSB1C3_pre_ccm">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>
Line 136: Line 212:
<td width="30%" id="pSB1C3_suf_pccH">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 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%" id="T7_frd_Ec_fwd">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%" id="T7_frd_Ec_rev">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%" id="pSB1C3_frd_pre">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%" id="pSB1C3_frd_suf">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%" id="pR6K_dcuB_short">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" id="Module2">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>
<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%" id="csoS3_can_fwd">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%" id="pSB1C3_pre_can">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">Integration of the <i>prkA</i> in the plasdmid pHnCBcsoS1D</th>
</tr>
</tr>
<tr>
<tr>
-
<td width="30%" id="csoS3_can_rev">csoS3_can_rev (41 bp)</td>
+
<td width="30%" id="pHnCBcsoS1D_fwd">pHnCBcsoS1D_fwd (44 bp)</td>
-
<td width="70%">5‘-TGCAGCGGCCGCTACTAGTATCATGCGGATGCAACCTCTTC-3‘</td>
+
<td width="70%">5‘-CGGTTGCCGCTAGCGTTTAAGTGAGCGGATAACAATTTCAG-3‘</td>
</tr>
</tr>
<tr>
<tr>
-
<td width="30%" id="pSB1C3_suf_can">pSB1C3_suf_can (46 bp)</td>
+
<td width="30%" id="pHNCBcsoS1D_rev">pHNCBcsoS1D_rev (45 bp)</td>
-
<td width="70%">5‘-TCGAAGAGGTTGCATCCGCATGATACTAGTAGCGGCCGCTGCAGTC-3‘</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>
<tr bgcolor="#D8D8D8">
<tr bgcolor="#D8D8D8">
-
   <th colspan="2" align="left">Shell associated protein</th>
+
   <th colspan="2" align="left">Shell associated protein of the carboxysome(<i>sap</i> respectively <i>csoS2</i>)</th>
</tr>
</tr>
<tr>
<tr>
Line 196: Line 287:
<td width="30%" id="csoS2_sap_1_rev">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>
<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%" id="csoS1D_fwd">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%" id="csoS1D_rev">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%" id="pSB1C3_suf_csoS1D">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%" id="pSB1C3_pre_csoS1D">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>
Line 252: Line 355:
</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>
 +
<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>
<tr>
<tr>
Line 261: Line 402:
<td width="30%" id="pSB1C3_suf_sRNApfkA">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">
Line 454: Line 603:
<td width="30%" id="zwf_purify_rev">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%" id="pSB1C3_pre_rbcLS">pSB1C3_pre_rbcLS (44 bp)</td>
 
-
<td width="70%">5‘-CATCTAGTATTTCTCCTCTTTCTCTAGAAGCGGCCGCGAATTCC-3‘</td>
 
-
</tr>
 
-
<tr>
 
-
<td width="30%" id="pSB1C3_suf_rbcLS">pSB1C3_suf_rbcLS (43 bp)</td>
 
-
<td width="70%">5‘-ATCGTCCGGGCCGCTACTAA-TACTAGTAGCGGCCGCTGCAGTC-3‘</td>
 
-
</tr>
 
-
<tr>
 
-
<td width="30%" id="pSB1C3_pre_ccbLS">pSB1C3_pre_ccbLS (44 bp)</td>
 
-
<td width="70%">5‘-CATCTAGTATTTCTCCTCTTTCTCTAGAAGCGGCCGCGAATTCC-3‘</td>
 
-
</tr>
 
-
<tr>
 
-
<td width="30%" id="pSB1C3_suf_ccbLS">pSB1C3_suf_ccbLS (43 bp)</td>
 
-
<td width="70%">5‘-TGGTTTACCGCGGCAACTAATACTAGTAGCGGCCGCTGCAGTC-3‘</td>
 
-
</tr>
 
-
<tr>
 
-
<td width="30%" id="pSB1C3_pre_prkA">pSB1C3_pre_prkA (43 bp)</td>
 
-
<td width="70%">5‘-ACACGATCTGGCTTGCTCATCTCTAGAAGCGGCCGCGAATTCC-3‘</td>
 
-
</tr>
 
-
<tr>
 
-
<td width="30%" id="pSB1C3_suf_prkA">pSB1C3_suf_prkA (43 bp)</td>
 
-
<td width="70%">5‘-CGGTTGCCGCTAGCGTTTAATACTAGTAGCGGCCGCTGCAGTC-3‘</td>
 
</tr>
</tr>
<tr bgcolor="#3fb498">
<tr bgcolor="#3fb498">

Revision as of 09:44, 26 August 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‘
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‘
pR6K_dcuB_short (22 bp) 5‘-AATTAACCCTCACTAAAGGGCG-3‘
NAPH/NAP+ 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‘
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‘
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‘
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‘
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‘
General primer
int_RBS_fw (1) (42 bp) 5‘-CTATAGGGAAAGAGGAGAAATGCATCACGGGAGATGCACTAG-3‘
RBS_int_rev (2) (44 bp) 5‘-CTAGTGCATCTCCCGTGATGCATTTCTCCTCTTTCCCTATAGTG-3‘
RBS_rev (3) (22 bp) 5‘-TTTCTCCTCTTTCCCTATAGTG-3‘
int_fw (4) (22 bp) 5‘-TGCATCACGGGAGATGCACTAG-3‘
int_pSB1C3_rev (5) (50 bp) 5‘-GACTGCAGCGGCCGCTACTAGTATCATTGAAGCTGCCACAAGGCAGGAAC-3‘
pSB1C3_int_fw (6) (46 bp) 5‘-CTGCCTTGTGGCAGCTTCAATGATACTAGTAGCGGCCGCTGCAGTC-3‘
SBPase (glpX)
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‘
fbaA
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‘
tktA
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‘
cg_pgk
cg_pgk_purify_fw (42 bp) 5‘-CTATAGGGAAAGAGGAGAAATATGGCTGTTAAGACCCTCAAG-3‘
cg_pgk_purify_rev (42 bp) 5‘-CTAGTGCATCTCCCGTGATGCACTGAGCGAGAATTGCAACGC-3‘
gapA
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‘
prkA
prkA_purify_fw (40 bp) 5‘-CTATAGGGAAAGAGGAGAAATATGAGCAAGCCAGATCGTG-3‘
prkA_purify_rev (42 bp) 5‘-CTAGTGCATCTCCCGTGATGCAAACGCTAGCGGCAACCGGTG-3‘
pykF
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‘
ldhA
ldhA_purify_fw (44 bp) 5‘-CTATAGGGAAAGAGGAGAAATATGAAACTCGCCGTTTATAGCAC-3‘
ldhA_purify_rev (42 bp) 5‘-CTAGTGCATCTCCCGTGATGCAAACCAGTTCGTTCGGGCAGG-3‘
glk
glk_purify_fw (44 bp) 5‘-CTATAGGGAAAGAGGAGAAATATGACAAAGTATGCATTAGTCGG-3‘
glk_purify_rev (43 bp) 5‘-CTAGTGCATCTCCCGTGATGCACAGAATGTGACCTAAGGTCTG-3‘
zwf
zwf_purify_fw (41 bp) 5‘-CTATAGGGAAAGAGGAGAAATATGGCGGTAACGCAAACAGC-3‘
zwf_purify_rev (45 bp) 5‘-CTAGTGCATCTCCCGTGATGCACTCAAACTCATTCCAGGAACGAC-3‘
Module 3
Backbone
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 (43 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‘
adhA
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‘
Biosafety