Template:SBB10 ConstructionFiles Chel sbb02: Difference between revisions
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Michel Nofal (talk | contribs) (New page: sbb02: N15 Protelomerase PCR MNF11-F/MNF12-R on pBca9523-Bca (945 bp, gp = A) PCR MNF13-F/MNF14-R on pBca9523-Bca (1033 bp, gp = B) ------------- PCR MNF11-F/MNF14-R on A + B (1925 bp...) |
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<pre> | |||
sbb02: N15 Protelomerase | sbb02: N15 Protelomerase | ||
PCR MNF11-F/MNF12-R on pBca9523- | PCR MNF11-F/MNF12-R on pBca9523-Bca1455 (945 bp, gp = A) | ||
PCR MNF13-F/MNF14-R on pBca9523- | PCR MNF13-F/MNF14-R on pBca9523-Bca1485 (1033 bp, gp = B) | ||
------------- | ------------- | ||
PCR MNF11-F/MNF14-R on A + B (1925 bp, EcoRI/BamHI) | PCR MNF11-F/MNF14-R on A + B (1925 bp, EcoRI/BamHI) | ||
Sub in pBjk2741-Bca1144 (EcoRI/BamHI, 910+2170, L) | Sub in pBjk2741-Bca1144 (EcoRI/BamHI, 910+2170, L) | ||
Product is pBjk2741-sbb02 {<protel!} | Product is pBjk2741-sbb02 {<protel!} | ||
------------- | ------------- | ||
MNF11-F ccagtGAATTCgtccAGATCTAGCAAGGTAAAAATCGGTGAGTTG | MNF11-F Forward PCR of Part 1 of sbb02 ccagtGAATTCgtccAGATCTAGCAAGGTAAAAATCGGTGAGTTG | ||
MNF12-R ggttacgcttttatcttcactgcgTTTTTTAGCTTGCCCTGAGAAATTAACCG | MNF12-R SOEing of Part1 of sbb02 ggttacgcttttatcttcactgcgTTTTTTAGCTTGCCCTGAGAAATTAACCG | ||
MNF13-F CGGTTAATTTCTCAGGGCAAGCTAAAAAAcgcagtgaagataaaagcgtaacc | MNF13-F SOEing of Part2 of sbb02 CGGTTAATTTCTCAGGGCAAGCTAAAAAAcgcagtgaagataaaagcgtaacc | ||
MNF14-R gcagtggatccTCAGCTGTAGTACGTTTCCCATGC | MNF14-R Reverse PCR of Part 2 of sbb02 gcagtggatccTCAGCTGTAGTACGTTTCCCATGC | ||
</pre> | |||
<blockquote style="background: seashell; border: 1px solid rgb(153, 153, 153); padding: 1em;"> | |||
==JCA Notes== | |||
*Have gtcc instead of atg in prefix location | |||
*Need to put part on Clotho | |||
*Otherwise correct | |||
*Internal oligos are longer than necessary, but would work | |||
*Due to similarity to other part, some oligos are being replaced: | |||
<pre> | |||
Construction of sbb02: N15 Protelomerase | |||
PCR MNF11-F/CA1472 on Bca1455 1-3 (569bp, gp=A) | |||
PCR ca1461/BDBn15001R on Bca1455 1-1 (762bp, gp=B) | |||
PCR BDBn15002F/BDBn15002R on Bca1485 2-1 (1023 bp, gp=C) | |||
-------------------------------------------------------------- | |||
PCR BDBn15001F/BDBn15002R on A+B+C (1924 bp, EcoRI/BamHI) | |||
Sub in pBjk2741-Bca1144 (EcoRI/BamHI, 910+2170, L) | |||
Product is pBjk2741-sbb01 {rbs.part!} | |||
MNF11-F Forward PCR of Part 1 of sbb02 | |||
ccagtGAATTCatgAGATCTAGCAAGGTAAAAATCGGTGAGTTG | |||
CA1461 GGAAAGGATTGCAGAAAAGAATAACCGCGAATACTTTTAACGCCTATATGA | |||
PCA assembly of protelomerasefiveprime (Bca1455) | |||
CA1472 TTGTTGGAATAACTTATAAAGATAGTCACGGCGCTCCTTCCAATCATCACT | |||
PCA assembly of protelomerasefiveprime (Bca1455) | |||
BDBn15001R Reverse retrieval and SOE-site introduciton on Bca1455 | |||
CGCTTTTATCTTCACTGCGTTTTTTAGCTTGCCCTGAG | |||
BDBn15002F Forward retrieval and SOE-site introduciton on Bca1485 | |||
CTCAGGGCAAGCTAAAAAACGCAGTGAAGATAAAAGCG | |||
BDBn15002R Reverse retrieval and BioBricking on Bca1485 | |||
CTGATggatccttaGCTGTAGTACGTTTCCCATGCGG | |||
part: | |||
GATCTAGCAAGGTAAAAATCGGTGAGTTGATCAACACGCTTGTGAATGAGGTAGAGGCCATTGATGCCTCAGACCGCCCACAAGGCGACAAAACGAAGAGAATTAAAGCCGCAGCCGCACGGTATAAGAACGCGTTATTTAATGATAAAAGAAAGTTCCGTGGGAAAGGATTGCAGAAAAGAATAACCGCGAATACTTTTAACGCCTATATGAGCAGGGCAAGAAAGCGGTTTGATGATAAATTACATCATAGCTTTGATAAAAATATTAATAAATTATCGGAAAAGTATCCTCTTTACAGCGAAGAATTATCTTCATGGCTTTCTATGCCTACGGCTAATATTCGCCAGCACATGTCATCGTTACAATCTAAATTGAAAGAAATAATGCCGCTTGCCGAAGAGTTATCAAATGTAAGAATAGGCTCTAAAGGCAGTGATGCAAAAATAGCAAGACTAATAAAAAAATATCCAGATTGGAGTTTTGCTCTTAGTGATTTAAACAGTGATGATTGGAAGGAGCGCCGTGACTATCTTTATAAGTTATTCCAACAAGGCTCTGCGTTGTTAGAAGAACTACACCAGCTCAAGGTCAACCATGAGGTTCTGTACCATCTCCAGCTAAGCCCTGCGGAGCGTACATCTATACAGCAACGATGGGCCGATGTTCTGCGCGAGAAGAAGCGTAATGTTGTGGTTATTGACTACCCAACATACATGCAGTCTATCTATGATATTTTGAATAATCCTGCGACTTTATTTAGTTTAAACACTCGTTCTGGAATGGCACCTTTGGCCTTTGCTCTGGCTGCGGTATCAGGGCGAAGAATGATTGAGATAATGTTTCAGGGTGAATTTGCCGTTTCAGGAAAGTATACGGTTAATTTCTCAGGGCAAGCTAAAAAACGCAGTGAAGATAAAAGCGTAACCAGAACGATTTATACTTTATGCGAAGCAAAATTATTCGTTGAATTATTAACAGAATTGCGTTCTTGCTCTGCTGCATCTGATTTCGATGAGGTTGTTAAAGGATATGGAAAGGATGATACAAGGTCTGAGAACGGCAGGATAAATGCTATTTTAGCAAAAGCATTTAACCCTTGGGTTAAATCATTTTTCGGCGATGACCGTCGTGTTTATAAAGATAGCCGCGCTATTTACGCTCGCATCGCTTATGAGATGTTCTTCCGCGTCGATCCACGGTGGAAAAACGTCGACGAGGATGTGTTCTTCATGGAGATTCTCGGACACGACGATGAGAACACCCAGCTGCACTATAAGCAGTTCAAGCTGGCCAACTTTTCCAGAACCTGGCGACCGGAAGTTGGGGATGAAAACACCAGGCTGGTGGCTCTCCAGAAACTGGACGATGAAATGCCAGGCTTTGCCAGAGGTGACGCTGGCGTCCGTCTCCATGAAACCGTTAAGCAGCTGGTGGAGCAGGACCCATCAGCAAAAATAACCAACAGCACTCTCCGGGCCTTTAAATTTAGCCCGACGATGATTAGCCGGTACCTGGAGTTTGCCGCTGATGCATTGGGGCAGTTCGTTGGCGAGAACGGGCAGTGGCAGCTCAAGATAGAGACACCTGCAATCGTCCTGCCTGATGAAGAATCCGTTGAGACCATCGACGAACCGGATGATGAGTCCCAAGACGACGAGCTGGATGAAGATGAAATTGAGCTCGACGAGGGTGGCGGCGATGAACCAACCGAAGAGGAAGGGCCAGAAGAACATCAGCCAACTGCTCTAAAACCCGTCTTCAAGCCTGCAAAAAATAACGGGGACGGAACGTACAAGATAGAGTTTGAATACGATGGAAAGCATTATGCCTGGTCCGGCCCCGCCGATAGCCCTATGGCCGCAATGCGATCCGCATGGGAAACGTACTACAGCTAAG | |||
</pre> | |||
</blockquote> | |||
__NOTOC__ |
Latest revision as of 16:48, 22 February 2010
sbb02: N15 Protelomerase PCR MNF11-F/MNF12-R on pBca9523-Bca1455 (945 bp, gp = A) PCR MNF13-F/MNF14-R on pBca9523-Bca1485 (1033 bp, gp = B) ------------- PCR MNF11-F/MNF14-R on A + B (1925 bp, EcoRI/BamHI) Sub in pBjk2741-Bca1144 (EcoRI/BamHI, 910+2170, L) Product is pBjk2741-sbb02 {<protel!} ------------- MNF11-F Forward PCR of Part 1 of sbb02 ccagtGAATTCgtccAGATCTAGCAAGGTAAAAATCGGTGAGTTG MNF12-R SOEing of Part1 of sbb02 ggttacgcttttatcttcactgcgTTTTTTAGCTTGCCCTGAGAAATTAACCG MNF13-F SOEing of Part2 of sbb02 CGGTTAATTTCTCAGGGCAAGCTAAAAAAcgcagtgaagataaaagcgtaacc MNF14-R Reverse PCR of Part 2 of sbb02 gcagtggatccTCAGCTGTAGTACGTTTCCCATGC
JCA Notes
- Have gtcc instead of atg in prefix location
- Need to put part on Clotho
- Otherwise correct
- Internal oligos are longer than necessary, but would work
- Due to similarity to other part, some oligos are being replaced:
Construction of sbb02: N15 Protelomerase PCR MNF11-F/CA1472 on Bca1455 1-3 (569bp, gp=A) PCR ca1461/BDBn15001R on Bca1455 1-1 (762bp, gp=B) PCR BDBn15002F/BDBn15002R on Bca1485 2-1 (1023 bp, gp=C) -------------------------------------------------------------- PCR BDBn15001F/BDBn15002R on A+B+C (1924 bp, EcoRI/BamHI) Sub in pBjk2741-Bca1144 (EcoRI/BamHI, 910+2170, L) Product is pBjk2741-sbb01 {rbs.part!} MNF11-F Forward PCR of Part 1 of sbb02 ccagtGAATTCatgAGATCTAGCAAGGTAAAAATCGGTGAGTTG CA1461 GGAAAGGATTGCAGAAAAGAATAACCGCGAATACTTTTAACGCCTATATGA PCA assembly of protelomerasefiveprime (Bca1455) CA1472 TTGTTGGAATAACTTATAAAGATAGTCACGGCGCTCCTTCCAATCATCACT PCA assembly of protelomerasefiveprime (Bca1455) BDBn15001R Reverse retrieval and SOE-site introduciton on Bca1455 CGCTTTTATCTTCACTGCGTTTTTTAGCTTGCCCTGAG BDBn15002F Forward retrieval and SOE-site introduciton on Bca1485 CTCAGGGCAAGCTAAAAAACGCAGTGAAGATAAAAGCG BDBn15002R Reverse retrieval and BioBricking on Bca1485 CTGATggatccttaGCTGTAGTACGTTTCCCATGCGG part: GATCTAGCAAGGTAAAAATCGGTGAGTTGATCAACACGCTTGTGAATGAGGTAGAGGCCATTGATGCCTCAGACCGCCCACAAGGCGACAAAACGAAGAGAATTAAAGCCGCAGCCGCACGGTATAAGAACGCGTTATTTAATGATAAAAGAAAGTTCCGTGGGAAAGGATTGCAGAAAAGAATAACCGCGAATACTTTTAACGCCTATATGAGCAGGGCAAGAAAGCGGTTTGATGATAAATTACATCATAGCTTTGATAAAAATATTAATAAATTATCGGAAAAGTATCCTCTTTACAGCGAAGAATTATCTTCATGGCTTTCTATGCCTACGGCTAATATTCGCCAGCACATGTCATCGTTACAATCTAAATTGAAAGAAATAATGCCGCTTGCCGAAGAGTTATCAAATGTAAGAATAGGCTCTAAAGGCAGTGATGCAAAAATAGCAAGACTAATAAAAAAATATCCAGATTGGAGTTTTGCTCTTAGTGATTTAAACAGTGATGATTGGAAGGAGCGCCGTGACTATCTTTATAAGTTATTCCAACAAGGCTCTGCGTTGTTAGAAGAACTACACCAGCTCAAGGTCAACCATGAGGTTCTGTACCATCTCCAGCTAAGCCCTGCGGAGCGTACATCTATACAGCAACGATGGGCCGATGTTCTGCGCGAGAAGAAGCGTAATGTTGTGGTTATTGACTACCCAACATACATGCAGTCTATCTATGATATTTTGAATAATCCTGCGACTTTATTTAGTTTAAACACTCGTTCTGGAATGGCACCTTTGGCCTTTGCTCTGGCTGCGGTATCAGGGCGAAGAATGATTGAGATAATGTTTCAGGGTGAATTTGCCGTTTCAGGAAAGTATACGGTTAATTTCTCAGGGCAAGCTAAAAAACGCAGTGAAGATAAAAGCGTAACCAGAACGATTTATACTTTATGCGAAGCAAAATTATTCGTTGAATTATTAACAGAATTGCGTTCTTGCTCTGCTGCATCTGATTTCGATGAGGTTGTTAAAGGATATGGAAAGGATGATACAAGGTCTGAGAACGGCAGGATAAATGCTATTTTAGCAAAAGCATTTAACCCTTGGGTTAAATCATTTTTCGGCGATGACCGTCGTGTTTATAAAGATAGCCGCGCTATTTACGCTCGCATCGCTTATGAGATGTTCTTCCGCGTCGATCCACGGTGGAAAAACGTCGACGAGGATGTGTTCTTCATGGAGATTCTCGGACACGACGATGAGAACACCCAGCTGCACTATAAGCAGTTCAAGCTGGCCAACTTTTCCAGAACCTGGCGACCGGAAGTTGGGGATGAAAACACCAGGCTGGTGGCTCTCCAGAAACTGGACGATGAAATGCCAGGCTTTGCCAGAGGTGACGCTGGCGTCCGTCTCCATGAAACCGTTAAGCAGCTGGTGGAGCAGGACCCATCAGCAAAAATAACCAACAGCACTCTCCGGGCCTTTAAATTTAGCCCGACGATGATTAGCCGGTACCTGGAGTTTGCCGCTGATGCATTGGGGCAGTTCGTTGGCGAGAACGGGCAGTGGCAGCTCAAGATAGAGACACCTGCAATCGTCCTGCCTGATGAAGAATCCGTTGAGACCATCGACGAACCGGATGATGAGTCCCAAGACGACGAGCTGGATGAAGATGAAATTGAGCTCGACGAGGGTGGCGGCGATGAACCAACCGAAGAGGAAGGGCCAGAAGAACATCAGCCAACTGCTCTAAAACCCGTCTTCAAGCCTGCAAAAAATAACGGGGACGGAACGTACAAGATAGAGTTTGAATACGATGGAAAGCATTATGCCTGGTCCGGCCCCGCCGATAGCCCTATGGCCGCAATGCGATCCGCATGGGAAACGTACTACAGCTAAG