Team:Bielefeld-CeBiTec/Notebook/Primer

From 2014.igem.org

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<td width="30%" id="pSB1C3_frd_suf">pSB1C3_frd_suf (47 bp)</td>
<td width="30%" id="pSB1C3_frd_suf">pSB1C3_frd_suf (47 bp)</td>
<td width="70%">5‘-CTGATTGGTGTCGTTACAATCTAATACTAGTAGCGGCCGCTGCAGTC-3‘</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>
<tr>
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<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>
 +
<td width="30%" id="ccm_con1">ccm_con1 (21 bp)</td>
 +
<td width="70%">5‘-ATTGGCATTGGCGGTTGGGTC-3‘</td>
</tr>
</tr>
<tr bgcolor="#3fb498">
<tr bgcolor="#3fb498">
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</tr>
</tr>
<tr>
<tr>
-
<td width="30%" id="int_RBS_fw">int_RBS_fw (1) (42 bp)</td>
+
<td width="30%" id="int_RBS_fw">int_RBS_fw (42 bp)</td>
<td width="70%">5‘-CTATAGGGAAAGAGGAGAAATGCATCACGGGAGATGCGCTAG-3‘</td>
<td width="70%">5‘-CTATAGGGAAAGAGGAGAAATGCATCACGGGAGATGCGCTAG-3‘</td>
</tr>
</tr>
<tr>
<tr>
-
<td width="30%" id="RBS_int_rev">RBS_int_rev (2) (44 bp)</td>
+
<td width="30%" id="RBS_int_rev">RBS_int_rev (44 bp)</td>
<td width="70%">5‘-CTAGCGCATCTCCCGTGATGCATTTCTCCTCTTTCCCTATAGTG-3‘</td>
<td width="70%">5‘-CTAGCGCATCTCCCGTGATGCATTTCTCCTCTTTCCCTATAGTG-3‘</td>
</tr>
</tr>
<tr>
<tr>
-
<td width="30%" id="RBS_rev">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%" id="int_fw">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%" id="int_pSB1C3_rev">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%" id="pSB1C3_int_fw">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">Expression of the SBPase (<i>glpX</i>)for the calvin cycle and for purification</th>
+
   <th colspan="2" align="left">Expression of the SBPase (<i>glpX</i>) for the calvin cycle and for purification</th>
</tr>
</tr>
<tr>
<tr>
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</tr>
</tr>
<tr bgcolor="#D8D8D8">
<tr bgcolor="#D8D8D8">
-
   <th colspan="2" align="left">Alcohol dehydrogenase (<i>adhA</i>) of <i>E. coli<i></th>
+
   <th colspan="2" align="left">Illegal restriction site between alsS and ilvC</th>
</tr>
</tr>
<tr>
<tr>
-
<td width="30%" id="fw-pSB1C3-Ec_adh">fw-pSB1C3-Ec_adh (59 bp)</td>
+
<td width="30%" id="rv_ilvC_alsS-new">rv_ilvC_alsS-new (72 bp)</td>
-
<td width="70%">5‘-CTGGAATTCGCGGCCGCTTCTAGAGAAAGAGGAGAAAATGCAGAACATCATCCGAAAAG-3‘</td>
+
<td width="70%">5‘-CAGTGTATTGAAGTAGTTAGCCATTTTCTCCTCTTTTTAGACAGTTTCCGTTTTCATGAGTTCCCCAAATTC-3‘</td>
</tr>
</tr>
<tr>
<tr>
-
<td width="30%" id="rv-pSB1C3-Ec_adh">rv-pSB1C3-Ec_adh (48 bp)</td>
+
<td width="30%" id="fw_alsS_ilvC-new">fw_alsS_ilvC-new (62 bp)</td>
-
<td width="70%">5‘-GACTGCAGCGGCCGCTACTAGTATTAGTGACGGAAATCAATCACCATG-3‘</td>
+
<td width="70%">5‘-CATGAAAACGGAAACTGTCTAAAAAGAGGAGAAAATGGCTAACTACTTCAATACACTGAATC-3‘</td>
-
</tr>
+
-
<tr>
+
-
<td width="30%" id="fw-adhA-pSBC1">fw-adhA-pSBC1 (48 bp)</td>
+
-
<td width="70%">5‘-CATGGTGATTGATTTCCGTCACTAATACTAGTAGCGGCCGCTGCAGTC-3‘</td>
+
-
</tr>
+
-
<tr>
+
-
<td width="30%" id="rv-adhA-pSBC1">rv-adhA-pSBC1 (51 bp)</td>
+
-
<td width="70%">5‘-CGGATGATGTTCTGCATTTTCTCCTCTTTCTCTAGAAGCGGCCGCGAATTC-3‘</td>
+
</tr>
</tr>
<tr bgcolor="#3fb498">
<tr bgcolor="#3fb498">
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<td width="30%" id="AB_kon_GFP_rev">AB_kon_GFP_rev (34 bp)</td>
<td width="30%" id="AB_kon_GFP_rev">AB_kon_GFP_rev (34 bp)</td>
<td width="70%">5‘-GCTACTAGTATTAAGCTACTAAAGCGTAGTTTTC-3‘</td>
<td width="70%">5‘-GCTACTAGTATTAAGCTACTAAAGCGTAGTTTTC-3‘</td>
-
</tr>
 
-
<tr bgcolor="#D8D8D8">
 
-
  <th colspan="2" align="left">LVA-tag for the D-alanin racemase</th>
 
-
</tr>
 
-
<tr>
 
-
<td width="30%" id="alr_LVA_fwd">alr_LVA_fwd (54 bp)</td>
 
-
<td width="70%">5‘-GCTACTAAAGCGTAGTTTTCGTCGTTTGCAGCGGTCGCGATGAAATACGTGGAT-3‘</td>
 
-
</tr>
 
-
<tr>
 
-
<td width="30%" id="alr_LVA_rev">alr_LVA_rev (52 bp)</td>
 
-
<td width="70%">5‘-AACGACGAAAACTACGCTTTAGTAGCTTAATATTTGATGCCTGGCTCTAGTA-3‘</td>
 
</tr>
</tr>

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"