Paris/Oligos

From 2007.igem.org

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== Constructions ==
+
{|
 +
|- style="background: #ccccff;"
 +
! colspan="5" style="background: #ccccff;" | Table of Oligos
 +
|- style="background: #ccccff; text-align: center;"
 +
|width=5%| Number
 +
|width=5%| Name
 +
|width=40%| Sequence
 +
|width=5%| Length
 +
|width=45%| Comments
 +
|- style="background: #cccccc;" 
 +
| style="background: #ccffcc;" |O1
 +
| Lox71-FtsZ-F
 +
| TTTGAATTCGCGGCCGCTTCTAGAGTACCGTTCGTATACGATACATT ATACGAAGTTATTAATAAACAGGCACAAATCGGAGAGAAACT
 +
| 89
 +
|
 +
|- style="background: #cccccc;"
 +
| style="background: #ccffcc;" | O2
 +
| FtsZ-R
 +
| TTTCTGCAGCGGCCGCTACTAGTATTTTTAATCAGCTTGCTTACGCA GGAAT
 +
| 49
 +
|
 +
|- style="background: #cccccc;"
 +
| style="background: #ccffcc;" | O3
 +
| Lox71-FtsA-F
 +
| TTTGAATTCGCGGCCGCTTCTAGAGTACCGTTCGTATACGATACATT ATACGAAGTTATTAATAATCAGGCACAGGCAGAACAACAATG
 +
| 89
 +
|
 +
|- style="background: #cccccc;"
 +
| style="background: #ccffcc;" | O4
 +
| DEcoR1-FtsZ-R
 +
| CGGTATTTACCGCGAAGAACTCAACACCTTCAATGCGCTC
 +
| 40
 +
|
 +
|- style="background: #cccccc;"
 +
| style="background: #ccffcc;" | O5
 +
| DEcoR1-FtsZ-F
 +
| TGCGCGAGCGCATTGAAGGTGTTGAGTTCTTCGCGGTAAA
 +
| 40
 +
|
 +
|- style="background: #cccccc;"
 +
| style="background: #ccffcc;" | O6
 +
| Lox66-DapAColi-F
 +
| TTTGAATTCGCGGCCGCTTCTAGAGATAACTTGGTATAGCATACATTATACG AACGGTAGTTTGCACAGAGGATGGCCCATGTT
 +
| 84
 +
|
 +
 +
|- style="background: #cccccc;"
 +
| style="background: #ccffcc;" | O7
 +
| DapAColi-R           
 +
| TTTCTGCAGCGGCCGCTACTAGTATTATTACAGCAAACCGGCATGCTTAAGC
 +
| 52
 +
|
-
* Lox71-FtsZ:
+
|- style="background: #cccccc;"
 +
| style="background: #ccffcc;" | O8
 +
| Lox66-DapASubtilis-F   
 +
| TTTGAATTCGCGGCCGCTTCTAGAGATAACTTGGTATAGCATACATTATACG AACGGTAATCATGATGAGGTGAATAAAATGAA
 +
| 84
 +
|
-
Lox71-FtsZ-F      TTTGAATTCGCGGCCGCTTCTAGAGTACCGTTCGTATACGATACATTATACGAAGTTATTAATAAACAGGCACAAATCGGAGAGAAACT 89     
+
|- style="background: #cccccc;"
 +
| style="background: #ccffcc;" | O9
 +
|DapASubtilis-R
 +
| TTTCTGCAGCGGCCGCTACTAGTATTATTACAGTTCGCTGATCGTGCTGCTT
 +
| 52
 +
|
-
FtsZ-R                TTTCTGCAGCGGCCGCTACTAGTATTTTTAATCAGCTTGCTTACGCAGGAAT 49
+
|- style="background: #cccccc;"
 +
| style="background: #ccffcc;" | O10
 +
| Lox71-KmR-F         
 +
| TTTCTCTAGAAGCGGCCGCGAATTCTACCGTTCGTATACGATACATTATACG AAGTTATCTGATCCTTCAACTCAGCAAAAGTTC
 +
| 85
 +
|
-
* Lox71-FtsA-FtsZ:
+
|- style="background: #cccccc;"
 +
| style="background: #ccffcc;" | O11
 +
| Lox66-KmR-R         
 +
| TTTTACTAGTAGCGGCCGCTGCAGTACCGTTCGTATAATGTATGCTATACCA AGTTATTTATTAGAAAAACTCATCGAGCATCAAA
 +
| 86
 +
|
-
Lox71-FtsA-F         TTTGAATTCGCGGCCGCTTCTAGAGTACCGTTCGTATACGATACATTATACGAAGTTATTAATAATCAGGCACAGGCAGAACAACAATG 89     
+
|- style="background: #cccccc;"
 +
| style="background: #ccffcc;" | O12
 +
| DPst1-DGAT-F                  
 +
| AGTGCTGTCGTATATGGCCCAGCAGGACTCAACATAATTT
 +
| 40
 +
|
-
* Deletion of Ecor1 site in FtsZ:
+
|- style="background: #cccccc;"
 +
| style="background: #ccffcc;" | O13
 +
| DPst1-DGAT-R         
 +
| AAATTATGTTGAGTCCTGCTGGGCCATATACGACAGCACT
 +
| 40
 +
|
-
DEcoR1-FtsZ-R        CGGTATTTACCGCGAAGAACTCAACACCTTCAATGCGCTC 40
 
-
DEcoR1-FtsZ-F         TGCGCGAGCGCATTGAAGGTGTTGAGTTCTTCGCGGTAAA 40
+
|- style="background: #cccccc;"
 +
| style="background: #ccffcc;" | O14
 +
| Lox66-F      
 +
| AATTCGCGGCCGCTTCTAGAGATAACTTGGTATAGCATACATTATACGAAC GGTATACTAGTAGCGGCCGCTGCA
 +
|
 +
|
 +
|- style="background: #cccccc;"
 +
| style="background: #ccffcc;" | O15
 +
| Lox66-R
 +
| GCGGCCGCTACTAGTATACCGTTCGTATAATGTATGCTATACCAAGTTATC TCTAGAAGCGGCCGCG
 +
|
 +
|
-
* Lox66-DapAColi:
+
|- style="background: #cccccc;"
 +
| style="background: #ccffcc;" | O16
 +
| Lox71-F
 +
| AATTCGCGGCCGCTTCTAGAGTACCGTTCGTATACGATACATTATACGAAG TTATTACTAGTAGCGGCCGCTGCA
 +
|
 +
|
-
Lox66-DapAColi-F      TTTGAATTCGCGGCCGCTTCTAGAGATAACTTGGTATAGCATACATTATACGAACGGTAGTTTGCACAGAGGATGGCCCATGTT 84   
+
|- style="background: #cccccc;"
 +
| style="background: #ccffcc;" | O17
 +
| Lox71-R
 +
| GCGGCCGCTACTAGTAATAACTTCGTATAATGTATCGTATACGAACGGTAC TCTAGAAGCGGCCGCG
 +
|
 +
|
-
DapAColi-R            TTTCTGCAGCGGCCGCTACTAGTATTATTACAGCAAACCGGCATGCTTAAGC 52
+
|- style="background: #cccccc;"
 +
| style="background: #ccffcc;" | O18
 +
| VF2
 +
| tgccacctgacgtctaagaa
 +
| 20
 +
| Forward Primer to Biobrick vectors (VF2) BBa_G00100
-
* Lox66-DapASubtilis:
+
|- style="background: #cccccc;"
 +
| style="background: #ccffcc;" | O19
 +
| VR
 +
| attaccgcctttgagtgagc
 +
| 20
 +
| Reverse Primer to Biobrick Vectors (VR) BBa_G00101
-
Lox66-DapASubtilis-F TTTGAATTCGCGGCCGCTTCTAGAGATAACTTGGTATAGCATACATTATACGAACGGTAATCATGATGAGGTGAATAAAATGAA 84     
+
|- style="background: #cccccc;"
 +
| style="background: #ccffcc;" | O20
 +
| RBS-DapAColi-F
 +
| TTTGAATTCGCGGCCGCTTCTAGAGATTAAAGAGGAGAAATACTAGATGT TCACGGGAAGTATTGTCGCGA
 +
| 71
 +
| Biobrick prefix + Strong RBS BBa_B0030 + 25 bp homology to DapA (Coli) from ATG
-
DapASubtilis-R        TTTCTGCAGCGGCCGCTACTAGTATTATTACAGTTCGCTGATCGTGCTGCTT 52
+
|- style="background: #cccccc;"
 +
| style="background: #ccffcc;" | O21
 +
| RBS-DapASubtilis-F
 +
| TTTGAATTCGCGGCCGCTTCTAGAGATTAAAGAGGAGAAATACTAGATGAA TTTCGGAAATGTGTCTACAG
 +
| 71
 +
| Biobrick prefix + Strong RBS BBa_B0030 + 25 bp homology to DapA (Subtilis) from ATG
 +
|- style="background: #cccccc;"
 +
| style="background: #ccffcc;" | O22
 +
| SEQFtsA-1F
 +
| GTCGTCCATACCGCGAAATC
 +
| 20
 +
|
-
* Lox71-KmR-Lox66:
+
|- style="background: #cccccc;"
 +
| style="background: #ccffcc;" | O23
 +
| SEQFtsA-2F
 +
| CGTGGAAGTGCCGAGCGTAG
 +
| 20
 +
|
-
Lox71-KmR-F          TTTCTCTAGAAGCGGCCGCGAATTCTACCGTTCGTATACGATACATTATACGAAGTTATCTGATCCTTCAACTCAGCAAAAGTTC 85 
+
|- style="background: #cccccc;"
 +
| style="background: #ccffcc;" | O24
 +
| SEQFtsZ-1F
 +
| GGTATCGCTGAACTGATTAC
 +
| 20
 +
|
-
Lox66-KmR-R          TTTTACTAGTAGCGGCCGCTGCAGTACCGTTCGTATAATGTATGCTATACCAAGTTATTTATTAGAAAAACTCATCGAGCATCAAA 86
+
|- style="background: #cccccc;"
 +
| style="background: #ccffcc;" | O25
 +
| SEQDapASub-1F
 +
| TTAAAGCAATTGCGGCAGAG
 +
| 20
 +
|
 +
|- style="background: #cccccc;"
 +
| style="background: #ccffcc;" | O26
 +
| SEQDapAColi-1F
 +
| CAATCGTCCGTCGCAAGAAG
 +
| 20
 +
|
 +
|- style="background: #cccccc;"
 +
| style="background: #ccffcc;" | O27
 +
| ForDGAT1
 +
| ATCCACGCTATGCGCCCATTAC
 +
| 22
 +
| Tm = 60°C
-
* Deletion of Pst1 site in DGAT:
+
|- style="background: #cccccc;"
 +
| style="background: #ccffcc;" | O28
 +
| RevDGAT1
 +
| GGGCTAATTTAGCCCTTTAGTT
 +
| 22
 +
| Tm = 50°C
-
DPst1-DGAT-F          AGTGCTGTCGTATATGGCCCAGCAGGACTCAACATAATTT 40
+
|- style="background: #cccccc;"
 +
| style="background: #ccffcc;" | O29
 +
| ForDGAT_BB1
 +
| AAAGAATTCGCGGCCGCTTCTAGAGATTAAAGAGGAGAAAATCCACGCTATG CGCCCATTAC
 +
| 62
 +
| Tm = 60/83°C
-
DPst1-DGAT-R         AAATTATGTTGAGTCCTGCTGGGCCATATACGACAGCACT 40
+
|- style="background: #cccccc;"
 +
| style="background: #ccffcc;" | O30
 +
| RevDGAT_BB1
 +
| CTGCAGCGGCCGCTACTAGTAGGGCTAATTTAGCCCTTTAGTT
 +
| 43
 +
| Tm = 50/74°C
 +
 
 +
|- style="background: #cccccc;"
 +
| style="background: #ccffcc;" | O31
 +
| DapAp-F
 +
| aattcgcggccgcttctagagAACAATCAGAACGGTTCTGTCTGCTTGCTTTT AATGCCATACCAAACGTACCATTGAGACACTTGTTTGCACAGAGGATGGta
 +
| 104
 +
|
 +
|- style="background: #cccccc;"
 +
| style="background: #ccffcc;" | O32
 +
| DapAp-R
 +
| ctagtaCCATCCTCTGTGCAAACAAGTGTCTCAATGGTACGTTTGGTATGGCA TTAAAAGCAAGCAGACAGAACCGTTCTGATTGTTctctagaagcggccgcg
 +
| 104
 +
|
 +
|- style="background: #cccccc;"
 +
| style="background: #ccffcc;" | O33
 +
| dDapA-F
 +
| CATACCAAACGTACCATTGAGACACTTGTTTGCACAGAGGAT GGCCCATGATTCCGGGGATCCGTCGACC
 +
| 70
 +
| [[Paris/July 27| more here]]
 +
|- style="background: #cccccc;"
 +
| style="background: #ccffcc;" | O34
 +
| dDapA-R
 +
| AAGCCATCAAATCTCCCTAAACTTTACAGCAAACCGGCATGC TTAAGCGCTGTAGGCTGGAGCTGCTTCG
 +
| 70
 +
| [[Paris/July 27| more here]]
 +
|- style="background: #cccccc;"
 +
| style="background: #ccffcc;" | O35
 +
| dFtsZ-F
 +
| GACGATGATTACGGCCTCAGGCGACAGGCACAAATCGGAGAGA AACTATGATTCCGGGGATCCGTCGACC
 +
| 70
 +
| [[Paris/July 27| more here]]
 +
|- style="background: #cccccc;"
 +
| style="background: #ccffcc;" | O36
 +
| dFtsZ-R
 +
| GAAACCCAAATTCCAGTCAATTCTTAATCAGCTTGCTTACGCA GGAATGCTGTAGGCTGGAGCTGCTTCG
 +
| 70
 +
| [[Paris/July 27| more here]]
 +
|- style="background: #cccccc;"
 +
| style="background: #ccffcc;" | O37
 +
| dDapAverif-F
 +
| ACAATCAGAACGGTTCTGTCTG
 +
| 22
 +
|
 +
|- style="background: #cccccc;"
 +
| style="background: #ccffcc;" | O38
 +
| dDapAverif-R
 +
| ATAGCGTGAGTCAGAACTACAG
 +
| 22
 +
|
 +
|- style="background: #cccccc;"
 +
| style="background: #ccffcc;" | O39
 +
| dFtsZverif-R
 +
| TCGCCAAATTACCTATCCAACC
 +
| 22
 +
|
 +
|- style="background: #cccccc;"
 +
| style="background: #ccffcc;" | O40
 +
| SEQ-mRFP-R
 +
| CGGACGACCTTCACCTTCAC
 +
|
 +
|
 +
|- style="background: #cccccc;"
 +
| style="background: #ccffcc;" | O41
 +
| M13(-20)-F
 +
|GTAAAACGACGGCCAG
 +
|
 +
|
 +
|- style="background: #cccccc;"
 +
| style="background: #ccffcc;" | O42
 +
| M13-R
 +
| CAGGAAACAGCTTATGAC
 +
|
 +
|
 +
|- style="background: #cccccc;"
 +
| style="background: #ccffcc;" | O43
 +
| Bfr-FtsK-F
 +
| TTATGGAAGAAGTCAAGCAGAGTAATCGTCTGGTTATTAAGACGCGCTAAT GCCACCTGACGTCTAAGAA
 +
| 70
 +
|
 +
|- style="background: #cccccc;"
 +
| style="background: #ccffcc;" | O44
 +
| Bfr-FtsK-R
 +
| CCAGGTTACCCCAGTACTGTTGCTGTATGGATTAACAGGAGTGTAATCAAA TTACCGCCTTTGAGTGAGC
 +
| 70
 +
|
 +
|- style="background: #cccccc;"
 +
| style="background: #ccffcc;" | O45
 +
| Aft-FtsK-F
 +
| AGGGGCACAACGGTAATCGTGAAGTGCTGGCCCCACCGCCGTTTGACTAAT GCCACCTGACGTCTAAGAA
 +
| 70
 +
|
 +
|- style="background: #cccccc;"
 +
| style="background: #ccffcc;" | O46
 +
| Aft-FtsK-R
 +
| GAAAATACTGAATTTTCAGTTGATAGAGTGCCGGATGCGACGCTACCGCGA TTACCGCCTTTGAGTGAGC
 +
| 70
 +
|
 +
|- style="background: #cccccc;"
 +
| style="background: #ccffcc;" | O47
 +
| PacI-pTet-F
 +
| TTATTAATTAAGAATTCGCGGCCGCATCTA
 +
| 30
 +
|
 +
|- style="background: #cccccc;"
 +
| style="background: #ccffcc;" | O48
 +
| XmaI-mrfpBB-R
 +
| GCATCCCGGGCAGCGGCCGCTTCTAGTATA
 +
| 30
 +
|
 +
|- style="background: #cccccc;"
 +
| style="background: #ccffcc;" | O49
 +
| PacI-pTet-F - oligold
 +
| GCTTATTAATTAAGAATTCGCGGCCGCATCTAG
 +
| 33
 +
|
 +
|- style="background: #cccccc;"
 +
| style="background: #ccffcc;" | O50
 +
| XmaI-mrfpBB-R - oligold
 +
| GGCATCCCGGGCAGCGGCCGCTTCTAGTATA
 +
| 31
 +
|
 +
|- style="background: #cccccc;"
 +
| style="background: #ccffcc;" | O51
 +
| Prefix-frt2-R
 +
| TTTCTGGAATTCGCGGCCGCATCTAGAATGGTTCGCTTCAGATCGTT
 +
| 47
 +
|
 +
|- style="background: #cccccc;"
 +
| style="background: #ccffcc;" | O52
 +
| SufixBB-frt1-F
 +
| AGGAAGCCTGCAGCGGCCGCTACTAGTA - TCTACAAACTCTGCTAGTTT - GTTTCGAAGTTCCTATACTT   
 +
final version with 5bp homology in frt1 & a mismatch at 14th bp (A>G) inabelling the SpeI site: GAAGCCTGCAGCGGCCGCTACTAGTAAACTCTGCTAGTTTCGAAGT
 +
Superfinal version with 12bp homology to each side from the mismatch (13th bp - A>G) : AGGAAGCCTGCAGCGGCCGCTACTAGTA - TCTACAAACTCTGCTAGTTTCGAAG
 +
| 48
 +
|
 +
|- style="background: #cccccc;"
 +
| style="background: #ccffcc;" | O53
 +
| SufixBB-T1T2frt1-F
 +
| ATTAAGCCTGCAGCGGCCGCTACTAGTAAATGGATCCCCCGGGGGCTG
 +
| 48
 +
|
 +
|}

Latest revision as of 23:46, 14 October 2007

Table of Oligos
Number Name Sequence Length Comments
O1 Lox71-FtsZ-F TTTGAATTCGCGGCCGCTTCTAGAGTACCGTTCGTATACGATACATT ATACGAAGTTATTAATAAACAGGCACAAATCGGAGAGAAACT 89
O2 FtsZ-R TTTCTGCAGCGGCCGCTACTAGTATTTTTAATCAGCTTGCTTACGCA GGAAT 49
O3 Lox71-FtsA-F TTTGAATTCGCGGCCGCTTCTAGAGTACCGTTCGTATACGATACATT ATACGAAGTTATTAATAATCAGGCACAGGCAGAACAACAATG 89
O4 DEcoR1-FtsZ-R CGGTATTTACCGCGAAGAACTCAACACCTTCAATGCGCTC 40
O5 DEcoR1-FtsZ-F TGCGCGAGCGCATTGAAGGTGTTGAGTTCTTCGCGGTAAA 40
O6 Lox66-DapAColi-F TTTGAATTCGCGGCCGCTTCTAGAGATAACTTGGTATAGCATACATTATACG AACGGTAGTTTGCACAGAGGATGGCCCATGTT 84
O7 DapAColi-R TTTCTGCAGCGGCCGCTACTAGTATTATTACAGCAAACCGGCATGCTTAAGC 52
O8 Lox66-DapASubtilis-F TTTGAATTCGCGGCCGCTTCTAGAGATAACTTGGTATAGCATACATTATACG AACGGTAATCATGATGAGGTGAATAAAATGAA 84
O9 DapASubtilis-R TTTCTGCAGCGGCCGCTACTAGTATTATTACAGTTCGCTGATCGTGCTGCTT 52
O10 Lox71-KmR-F TTTCTCTAGAAGCGGCCGCGAATTCTACCGTTCGTATACGATACATTATACG AAGTTATCTGATCCTTCAACTCAGCAAAAGTTC 85
O11 Lox66-KmR-R TTTTACTAGTAGCGGCCGCTGCAGTACCGTTCGTATAATGTATGCTATACCA AGTTATTTATTAGAAAAACTCATCGAGCATCAAA 86
O12 DPst1-DGAT-F AGTGCTGTCGTATATGGCCCAGCAGGACTCAACATAATTT 40
O13 DPst1-DGAT-R AAATTATGTTGAGTCCTGCTGGGCCATATACGACAGCACT 40


O14 Lox66-F AATTCGCGGCCGCTTCTAGAGATAACTTGGTATAGCATACATTATACGAAC GGTATACTAGTAGCGGCCGCTGCA
O15 Lox66-R GCGGCCGCTACTAGTATACCGTTCGTATAATGTATGCTATACCAAGTTATC TCTAGAAGCGGCCGCG
O16 Lox71-F AATTCGCGGCCGCTTCTAGAGTACCGTTCGTATACGATACATTATACGAAG TTATTACTAGTAGCGGCCGCTGCA
O17 Lox71-R GCGGCCGCTACTAGTAATAACTTCGTATAATGTATCGTATACGAACGGTAC TCTAGAAGCGGCCGCG
O18 VF2 tgccacctgacgtctaagaa 20 Forward Primer to Biobrick vectors (VF2) BBa_G00100
O19 VR attaccgcctttgagtgagc 20 Reverse Primer to Biobrick Vectors (VR) BBa_G00101
O20 RBS-DapAColi-F TTTGAATTCGCGGCCGCTTCTAGAGATTAAAGAGGAGAAATACTAGATGT TCACGGGAAGTATTGTCGCGA 71 Biobrick prefix + Strong RBS BBa_B0030 + 25 bp homology to DapA (Coli) from ATG
O21 RBS-DapASubtilis-F TTTGAATTCGCGGCCGCTTCTAGAGATTAAAGAGGAGAAATACTAGATGAA TTTCGGAAATGTGTCTACAG 71 Biobrick prefix + Strong RBS BBa_B0030 + 25 bp homology to DapA (Subtilis) from ATG
O22 SEQFtsA-1F GTCGTCCATACCGCGAAATC 20
O23 SEQFtsA-2F CGTGGAAGTGCCGAGCGTAG 20
O24 SEQFtsZ-1F GGTATCGCTGAACTGATTAC 20
O25 SEQDapASub-1F TTAAAGCAATTGCGGCAGAG 20
O26 SEQDapAColi-1F CAATCGTCCGTCGCAAGAAG 20
O27 ForDGAT1 ATCCACGCTATGCGCCCATTAC 22 Tm = 60°C
O28 RevDGAT1 GGGCTAATTTAGCCCTTTAGTT 22 Tm = 50°C
O29 ForDGAT_BB1 AAAGAATTCGCGGCCGCTTCTAGAGATTAAAGAGGAGAAAATCCACGCTATG CGCCCATTAC 62 Tm = 60/83°C
O30 RevDGAT_BB1 CTGCAGCGGCCGCTACTAGTAGGGCTAATTTAGCCCTTTAGTT 43 Tm = 50/74°C
O31 DapAp-F aattcgcggccgcttctagagAACAATCAGAACGGTTCTGTCTGCTTGCTTTT AATGCCATACCAAACGTACCATTGAGACACTTGTTTGCACAGAGGATGGta 104
O32 DapAp-R ctagtaCCATCCTCTGTGCAAACAAGTGTCTCAATGGTACGTTTGGTATGGCA TTAAAAGCAAGCAGACAGAACCGTTCTGATTGTTctctagaagcggccgcg 104
O33 dDapA-F CATACCAAACGTACCATTGAGACACTTGTTTGCACAGAGGAT GGCCCATGATTCCGGGGATCCGTCGACC 70 more here
O34 dDapA-R AAGCCATCAAATCTCCCTAAACTTTACAGCAAACCGGCATGC TTAAGCGCTGTAGGCTGGAGCTGCTTCG 70 more here
O35 dFtsZ-F GACGATGATTACGGCCTCAGGCGACAGGCACAAATCGGAGAGA AACTATGATTCCGGGGATCCGTCGACC 70 more here
O36 dFtsZ-R GAAACCCAAATTCCAGTCAATTCTTAATCAGCTTGCTTACGCA GGAATGCTGTAGGCTGGAGCTGCTTCG 70 more here
O37 dDapAverif-F ACAATCAGAACGGTTCTGTCTG 22
O38 dDapAverif-R ATAGCGTGAGTCAGAACTACAG 22
O39 dFtsZverif-R TCGCCAAATTACCTATCCAACC 22
O40 SEQ-mRFP-R CGGACGACCTTCACCTTCAC
O41 M13(-20)-F GTAAAACGACGGCCAG
O42 M13-R CAGGAAACAGCTTATGAC
O43 Bfr-FtsK-F TTATGGAAGAAGTCAAGCAGAGTAATCGTCTGGTTATTAAGACGCGCTAAT GCCACCTGACGTCTAAGAA 70
O44 Bfr-FtsK-R CCAGGTTACCCCAGTACTGTTGCTGTATGGATTAACAGGAGTGTAATCAAA TTACCGCCTTTGAGTGAGC 70
O45 Aft-FtsK-F AGGGGCACAACGGTAATCGTGAAGTGCTGGCCCCACCGCCGTTTGACTAAT GCCACCTGACGTCTAAGAA 70
O46 Aft-FtsK-R GAAAATACTGAATTTTCAGTTGATAGAGTGCCGGATGCGACGCTACCGCGA TTACCGCCTTTGAGTGAGC 70
O47 PacI-pTet-F TTATTAATTAAGAATTCGCGGCCGCATCTA 30
O48 XmaI-mrfpBB-R GCATCCCGGGCAGCGGCCGCTTCTAGTATA 30
O49 PacI-pTet-F - oligold GCTTATTAATTAAGAATTCGCGGCCGCATCTAG 33
O50 XmaI-mrfpBB-R - oligold GGCATCCCGGGCAGCGGCCGCTTCTAGTATA 31
O51 Prefix-frt2-R TTTCTGGAATTCGCGGCCGCATCTAGAATGGTTCGCTTCAGATCGTT 47
O52 SufixBB-frt1-F AGGAAGCCTGCAGCGGCCGCTACTAGTA - TCTACAAACTCTGCTAGTTT - GTTTCGAAGTTCCTATACTT

final version with 5bp homology in frt1 & a mismatch at 14th bp (A>G) inabelling the SpeI site: GAAGCCTGCAGCGGCCGCTACTAGTAAACTCTGCTAGTTTCGAAGT Superfinal version with 12bp homology to each side from the mismatch (13th bp - A>G) : AGGAAGCCTGCAGCGGCCGCTACTAGTA - TCTACAAACTCTGCTAGTTTCGAAG

48
O53 SufixBB-T1T2frt1-F ATTAAGCCTGCAGCGGCCGCTACTAGTAAATGGATCCCCCGGGGGCTG 48
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