"
Team:Waterloo/Notebook/Oligos
From 2009.igem.org
(Difference between revisions)
| Line 13: | Line 13: | ||
|amplifies second half of alpha gal gene, adds XbaI site (rev) || H2-XbaI-R || ATGTCTAGAGGTCAGCAGCGTCTGT || | |amplifies second half of alpha gal gene, adds XbaI site (rev) || H2-XbaI-R || ATGTCTAGAGGTCAGCAGCGTCTGT || | ||
|- | |- | ||
| - | |Amplifes any biobrick, adds EcoRV end for ligation into EcoRV blunt ends (fwd) || Prefix-EcoRV-F || | + | |Amplifes any biobrick, adds EcoRV end for ligation into EcoRV blunt ends (fwd) || Prefix-EcoRV-F || ATCGAATTCGCGGCCGCTTCTAG || |
|- | |- | ||
| - | |Amplifes any biobrick, adds EcoRV end for ligation into EcoRV blunt ends (rev) || Suffix-EcoRV-R || | + | |Amplifes any biobrick, adds EcoRV end for ligation into EcoRV blunt ends (rev) || Suffix-EcoRV-R || ATCCTGCAGCGGCCGCTACTAGTA || |
|- | |- | ||
|B0034 RBS primer, use with VF2 (fwd) || Prefix-RBS || GGCCGCTTCTAGAGAAAGAGG || 21 | |B0034 RBS primer, use with VF2 (fwd) || Prefix-RBS || GGCCGCTTCTAGAGAAAGAGG || 21 | ||
| Line 23: | Line 23: | ||
|ordered to add Plac and RBS to the cI ORF (project 2008) || Plac-RBS-F || GAATTCGCGGCCGCTTCTAGAGAATTGTGAGCGGATAACAATTGACATTGTGAGCGGATAA CAAGATACTGAGCACATACTAGAGAAAGAGGAGAAATACTAG || 103 | |ordered to add Plac and RBS to the cI ORF (project 2008) || Plac-RBS-F || GAATTCGCGGCCGCTTCTAGAGAATTGTGAGCGGATAACAATTGACATTGTGAGCGGATAA CAAGATACTGAGCACATACTAGAGAAAGAGGAGAAATACTAG || 103 | ||
|- | |- | ||
| + | |amplifies c31 int, adds RBS, Prefix, changes start codon to ATG, changes G to A maintaining glutamic acid codon and eliminating EcoRI site (fwd) || Int-BBa-F || GAATTCGCGGCCGCTTCTAGAGAAAGAGGAGAAATACTAGATGGACACGTACGCGGGTGCTTACGACCGTCAGTCGCGCGAGCGCGAAAATTCG || | ||
| + | |- | ||
| + | |amplifies c31 int, adds suffix, changes stop codon to double TAA (rev) || Int-DSC-R || CTGCAGCGGCCGCTACTAGTATTATTAcgccgctacgtcttccgtg || | ||
| + | |- | ||
| + | |amplifies T7 Gene 6 exonuclease, adds ATG start codon, adds BBa prefix (fwd2) || T7G6-F2 || GAATTCGCGGCCGCTTCTAGATGGCACTTCTTGACCTTAAAC || | ||
| + | |- | ||
| + | |amplifies T7 Gene 6 exonuclease, adds double TAA stop codon, adds BBa suffix (rev2) || T7G6-R2 || CTGCAGCGGCCGCTACTAGTATTATTACGGTCTCCACAGGTAAATCTC || | ||
| + | |- | ||
| + | |amplifies T7 G6 makes product for cross over PCR with PmeI cross over PCR product || T7g6-RXO-RBS || CTAGTATTTCTCCTCTTTCTCTAGTATTATTAcggtctccacaggtaaatctc || | ||
| + | |- | ||
| + | |||
| + | ||- | ||
| + | |||
|BBa sequencing/Colony PCR primer (fwd) || VF2 || TGCCACCTGACGTCTAAGAA || 20 | |BBa sequencing/Colony PCR primer (fwd) || VF2 || TGCCACCTGACGTCTAAGAA || 20 | ||
Revision as of 06:19, 20 October 2009
| Description | Oligo Name | Sequence | Length | |
|---|---|---|---|---|
| amplifies first half of E. coli alpha gal gene adds SalI REN site (fwd) | HI-SalI-F | CATGTCGACATGATGTCTGCACCCAAAATT | ||
| amplifies first half of alpha gal gene adds BamHI REN site (rev) | HI-BamHI-attP-R | ATGGGATCCCCCCCAACTGAGAGAACTCAAAGGTTACCCCAGTTGGGGGTAACGCAGCGTAGCTGG | ||
| amplifies second half of alpha gal gene, adds KpnI site (fwd) | H2-KpnI-attP-F | CATGGTACCCCCCCAACTGAGAGAACTCAAAGGTTACCCCAGTTGGGGGCGTTTTACCTGGAGCTGG | ||
| amplifies second half of alpha gal gene, adds XbaI site (rev) | H2-XbaI-R | ATGTCTAGAGGTCAGCAGCGTCTGT | ||
| Amplifes any biobrick, adds EcoRV end for ligation into EcoRV blunt ends (fwd) | Prefix-EcoRV-F | ATCGAATTCGCGGCCGCTTCTAG | ||
| Amplifes any biobrick, adds EcoRV end for ligation into EcoRV blunt ends (rev) | Suffix-EcoRV-R | ATCCTGCAGCGGCCGCTACTAGTA | ||
| B0034 RBS primer, use with VF2 (fwd) | Prefix-RBS | GGCCGCTTCTAGAGAAAGAGG | 21 | |
| B0034 RBS primer, use with VR (rev) | Suffix-RBS | TTTCTCCTCTTTCTCTAGATGCGG | 24 | |
| ordered to add Plac and RBS to the cI ORF (project 2008) | Plac-RBS-F | GAATTCGCGGCCGCTTCTAGAGAATTGTGAGCGGATAACAATTGACATTGTGAGCGGATAA CAAGATACTGAGCACATACTAGAGAAAGAGGAGAAATACTAG | 103 | |
| amplifies c31 int, adds RBS, Prefix, changes start codon to ATG, changes G to A maintaining glutamic acid codon and eliminating EcoRI site (fwd) | Int-BBa-F | GAATTCGCGGCCGCTTCTAGAGAAAGAGGAGAAATACTAGATGGACACGTACGCGGGTGCTTACGACCGTCAGTCGCGCGAGCGCGAAAATTCG | ||
| amplifies c31 int, adds suffix, changes stop codon to double TAA (rev) | Int-DSC-R | CTGCAGCGGCCGCTACTAGTATTATTAcgccgctacgtcttccgtg | ||
| amplifies T7 Gene 6 exonuclease, adds ATG start codon, adds BBa prefix (fwd2) | T7G6-F2 | GAATTCGCGGCCGCTTCTAGATGGCACTTCTTGACCTTAAAC | ||
| amplifies T7 Gene 6 exonuclease, adds double TAA stop codon, adds BBa suffix (rev2) | T7G6-R2 | CTGCAGCGGCCGCTACTAGTATTATTACGGTCTCCACAGGTAAATCTC | ||
| amplifies T7 G6 makes product for cross over PCR with PmeI cross over PCR product | T7g6-RXO-RBS | CTAGTATTTCTCCTCTTTCTCTAGTATTATTAcggtctccacaggtaaatctc | ||
| -
| BBa sequencing/Colony PCR primer (fwd) | VF2 | TGCCACCTGACGTCTAAGAA | 20 |
| BBa sequencing/Colony PCR primer (rev) | VR | ATTACCGCCTTTGAGTGAGC | 20 | |
| T7 gene 5 primer, 3'-5' exonuclease, adds BBa prefix (fwd) | T7gene5-F | CAATGGAATTCGCGGCCGCTTCTAGATGATCGTTTCTGACATCGAA | 46 | |
| T7 gene 5 primer, 3'-5' exonuclease, adds BBa suffix (rev) | T7gene5-R | CAATTTCTGCAGCGGCCGCTACTAGTATCAGTGGCAAATCGCCCAATTAGG | 51 | |
| Gene 6 ORF from T7 phage (fwd; Prefix adapter) | T7gene6-F | CAATGGAATTCGCGGCCGCTTCTAGATGGCACTTCTTGACCTTAAA | 46 | |
| Gene 6 ORF from T7 phage (rev; Suffix adapter) | T7gene6-R | CAATTTCTGCAGCGGCCGCTACTAGTACTACGGTCTCCACAGG | 43 | |
| PmeIR ORF (fwd) | PmeIR-F | ATCCGGAATTCGCGGCGCTTTCTAGATGACAACAAACTCCCCCTCAGACG | 50 | |
| PmeIR ORF (rev) | PmelR-R | CGCAAAACTGCAGCGGCCGCTACTAGTATCAAACCGTACTTCGCCCCCGTTG | 52 | |
| pRecA oligo (sense strand; produces BioBrick-compatible EcoRI/SpeI overhangs when annealed to recAR) | recAF | AATTCGCGGCCGCTTCTAGAGTACAAAACACTTGATACTGTATATATATACAGTATAATTGCTTCAACATA | 71 | |
| pRecA oligo (antisense strand; produces BioBrick-compatible EcoRI/SpeI overhangs when annealed to recAR) | recAR | CTAGTATGTTGAAGCAATTATACTGTATATATATACAGTATCAAGTGTTTTGTACTCTAGAAGCGGCCGCG | 71 | |
| {B0034}-{T7gene6 ORF} (fwd; for adding {Prefix-RBS} onto the gene 6 ORF by PCR) | RBST7g6-F | GAATTCCGGCCGCTTCTAGAGAAAGAGGAGAAATACTAGATGGCACTTCTTGACCTTAAAC | 61 | |
| T7gene6 ORF (rev) | T7gs-R | CTGCAGCGGCCGCTACTAGTATTATTACGGTCTCCACAGGTAAATCTC | 48 | |
| T7gene6 ORF rev; for joining to PmeI with crossover PCR | T7g6-RXO-RBS | CTAGTATTTCTCCTCTTTCTCTAGTATTATTACGGTCTCCACAGGTAAATCTC | 53 | |
| PmeI ORF (fwd; for adding prefix-RBS) | RBSpmeI-F | GAATTCCGGCCGCTTCTAGAGAAAGAGGAGAAATACTAGATGACAACAAACTCCCCCTCAG | 61 | |
| PmeI ORF (fwd; for adding prefix-RBS and joining to T7gene6 by crossover PCR) | RBSpmeI-FXO | TACTAGAGAAAGAGGAGAAATACTAGATGACAACAAACTCCCCCTCAG | 48 | |
| PmeI ORF (rev; TAATAA stop codon-suffix) | pmeI-R | CTGCAGCGGCCGCTACTAGTATTATTAAACCGTACTTCGCCCCCG | 45 | |
| Primer for part R0011 (Plac-PL hybrid promoter; fwd) | R0011-F | GAATTCCGGCCGCTTCTAGAGAATTGTGAGCGGATAAC | 38 | |
| Primer for part R0011 (Plac-PL hybrid promoter; rev for joining to ORFs with RBS by crossover PCR) | R0011RBS-RXO | CTAGTATTTCTCCTCTTTCTCTAGTATGTGCTCAGTATCTTGTTATC | 47 | |
| CI ORF (fwd; prefix-RBS) | RBSC0051-F | GAATTCCGGCCGCTTCTAGAGAAAGAGGAGAAATACTAGATGAGCACAAAAAAGAAACC | 59 | |
| CI ORF (fwd; prefix-RBS for joining to R0011 promoter by crossover PCR) | RBSC0051-FXO | TACTAGAGAAAGAGGAGAAATACTAGATGAGCACAAAAAAGAAACC | 46 | |
| CI ORF (rev; suffix adapter) | C0051-R | CTGCAGCGGCCGCTACTAGTATTATTAAGCTACTAAAGCG | 40 | |
| CI ORF (rev; for joining to transcriptional terminator (b0015) by crossover PCR) | C0051-RXO | GATGCCTGGCTCTAGTATTATTAAGCTACTAAAGCG | 36 | |
| Transcriptional terminator (B0015; fwd; for joining to ORFs by crossover PCR) | B0015-FXO | TAATAATACTAGAGCCAGGCATCAAATAAAACG | 33 | |
| Transcriptional terminator (B0015; rev) | B0015-R | CTGCAGCGGCCGCTACTAGTATATAAACGCAGAAAGGCC | 39 | |
| RFP ORF (fwd; prefix-RBS) | RFP-RBS-F | GAATTCCGGCCGCTTCTAGAGAAAGAGGAGAAATACTAGATGGCTTCCTCCGAAGACG | 58 | |
| RFP ORF (fwd; prefix-RBS for joining to transcriptional terminator by crossover PCR ) | RFP-RXO | GATGCCTGGCTCTAGTATTATTAAGCACCGGTGG | 34 |
