pTYB11
At a Glance
pTYB11; Image provided by NEB.
<protect>
| Type |
Cloning & Expression |
|---|---|
| Origin of Replication |
pMB1 |
| Copy # |
~20 per cell |
| Markers |
Ampicillin |
| Link to Sequence | |
| Features |
lacI
[1] |
| Accession Number |
|
| edit table |
</protect>
Notes
The pTYB1 is an E. coli vector created for fusion protein expression and purification. A target gene can be inserted into the the ORF by use of the MCS downstream of the Sce VMA intein. The SapI site in the MCS is responsible for cloning the 5' end of the inserted gene. The chitin-binding domain(CBD) within the intein gene results in a intein-protein with chitin beads that facilitate protein purification. Release of the protein is induced by the thiol cleavage activity of the intein.
History
pTYB11 is derived from E. coli plamid pBR322, its chitin-binding domain(CBD) is isolated from B. circulans, and the VMA intein from S. cerevisiae.
Sources
References
See Help:References for how to manage references in EcoliWiki.
- ↑ 1.0 1.1 Dubendorff, JW & Studier, FW (1991) Controlling basal expression in an inducible T7 expression system by blocking the target T7 promoter with lac repressor. J. Mol. Biol. 219 45-59 PubMed
- ↑ 2.0 2.1 Chong, S et al. (1998) Utilizing the C-terminal cleavage activity of a protein splicing element to purify recombinant proteins in a single chromatographic step. Nucleic Acids Res. 26 5109-15 PubMed
- ↑ 3.0 3.1 Chong, S et al. (1997) Single-column purification of free recombinant proteins using a self-cleavable affinity tag derived from a protein splicing element. Gene 192 271-81 PubMed
- ↑ Chong, S et al. (1998) Modulation of protein splicing of the Saccharomyces cerevisiae vacuolar membrane ATPase intein. J. Biol. Chem. 273 10567-77 PubMed
Sequence
1 AGAAGGCCAT CCTGACGGAT GGCCTTTTTG CGTTTCTACA AACTCTTTTT
51 GTTTATTTTT CTAAATACAT TCAAATATGT ATCCGCTCAT GAGACAATAA
101 CCCTGATAAA TGCTTCAATA ATATTGAAAA AGGAAGAGTA TGAGTATTCA
151 ACATTTCCGT GTCGCCCTTA TTCCCTTTTT TGCGGCATTT TGCCTTCCTG
201 TTTTTGCTCA CCCAGAAACG CTGGTGAAAG TAAAAGATGC TGAAGATCAG
251 TTGGGTGCAC GAGTGGGTTA CATCGAACTG GATCTCAACA GCGGTAAGAT
301 CCTTGAGAGT TTTCGCCCCG AAGAACGTTC TCCAATGATG AGCACTTTTA
351 AAGTTCTGCT ATGTGGCGCG GTATTATCCC GTGTTGACGC CGGGCAAGAG
401 CAACTCGGTC GCCGCATACA CTATTCTCAG AATGACTTGG TTGAGTACTC
451 ACCAGTCACA GAAAAGCATC TTACGGATGG CATGACAGTA AGAGAATTAT
501 GCAGTGCTGC CATAACCATG AGTGATAACA CTGCGGCCAA CTTACTTCTG
551 ACAACGATCG GAGGACCGAA GGAGCTAACC GCTTTTTTGC ACAACATGGG
601 GGATCATGTA ACTCGCCTTG ATCGTTGGGA ACCGGAGCTG AATGAAGCCA
651 TACCAAACGA CGAGCGTGAC ACCACGATGC CTGTAGCAAT GGCAACAACG
701 TTGCGCAAAC TATTAACTGG CGAACTACTT ACTCTAGCTT CCCGGCAACA
751 ATTAATAGAC TGGATGGAGG CGGATAAAGT TGCAGGACCA CTTCTGCGCT
801 CGGCCCTTCC GGCTGGCTGG TTTATTGCTG ATAAATCTGG AGCCGGTGAG
851 CGTGGGTCTC GCGGTATCAT TGCAGCACTG GGGCCAGATG GTAAGCCCTC
901 CCGTATCGTA GTTATCTACA CGACGGGGAG TCAGGCAACT ATGGATGAAC
951 GAAATAGACA GATCGCTGAG ATAGGTGCCT CACTGATTAA GCATTGGTAA
1001 CTGTCAGACC AAGTTTACTC ATATATACTT TAGATTGATT TACCCCGGTT
1051 GATAATCAGA AAAGCCCCAA AAACAGGAAG ATTGTATAAG CAAATATTTA
1101 AATTGTAAAC GTTAATATTT TGTTAAAATT CGCGTTAAAT TTTTGTTAAA
1151 TCAGCTCATT TTTTAACCAA TAGGCCGAAA TCGGCAAAAT CCCTTATAAA
1201 TCAAAAGAAT AGCCCGAGAT AGGGTTGAGT GTTGTTCCAG TTTGGAACAA
1251 GAGTCCACTA TTAAAGAACG TGGACTCCAA CGTCAAAGGG CGAAAAACCG
1301 TCTATCAGGG CGATGGCCCA CTACGTGAAC CATCACCCAA ATCAAGTTTT
1351 TTGGGGTCGA GGTGCCGTAA AGCACTAAAT CGGAACCCTA AAGGGAGCCC
1401 CCGATTTAGA GCTTGACGGG GAAAGCCGGC GAACGTGGCG AGAAAGGAAG
1451 GGAAGAAAGC GAAAGGAGCG GGCGCTAGGG CGCTGGCAAG TGTAGCGGTC
1501 ACGCTGCGCG TAACCACCAC ACCCGCCGCG CTTAATGCGC CGCTACAGGG
1551 CGCGTAAAAG GATCTAGGTG AAGATCCTTT TTGATAATCT CATGACCAAA
1601 ATCCCTTAAC GTGAGTTTTC GTTCCACTGA GCGTCAGACC CCGTAGAAAA
1651 GATCAAAGGA TCTTCTTGAG ATCCTTTTTT TCTGCGCGTA ATCTGCTGCT
1701 TGCAAACAAA AAAACCACCG CTACCAGCGG TGGTTTGTTT GCCGGATCAA
1751 GAGCTACCAA CTCTTTTTCC GAAGGTAACT GGCTTCAGCA GAGCGCAGAT
1801 ACCAAATACT GTCCTTCTAG TGTAGCCGTA GTTAGGCCAC CACTTCAAGA
1851 ACTCTGTAGC ACCGCCTACA TACCTCGCTC TGCTAATCCT GTTACCAGTG
1901 GCTGCTGCCA GTGGCGATAA GTCGTGTCTT ACCGGGTTGG ACTCAAGACG
1951 ATAGTTACCG GATAAGGCGC AGCGGTCGGG CTGAACGGGG GGTTCGTGCA
2001 CACAGCCCAG CTTGGAGCGA ACGACCTACA CCGAACTGAG ATACCTACAG
2051 CGTGAGCTAT GAGAAAGCGC CACGCTTCCC GAAGGGAGAA AGGCGGACAG
2101 GTATCCGGTA AGCGGCAGGG TCGGAACAGG AGAGCGCACG AGGGAGCTTC
2151 CAGGGGGAAA CGCCTGGTAT CTTTATAGTC CTGTCGGGTT TCGCCACCTC
2201 TGACTTGAGC GTCGATTTTT GTGATGCTCG TCAGGGGGGC GGAGCCTATG
2251 GAAAAACGCC AGCAACGCGG CCTTTTTACG GTTCCTGGCC TTTTGCTGGC
2301 CTTTTGCTCA CATGTTCTTT CCTGCGTTAT CCCCTGATTC TGTGGATAAC
2351 CGTATTACCG CCTTTGAGTG AGCTGATACC GCTCGCCGCA GCCGAACGAC
2401 CGAGCGCAGC GAGTCAGTGA GCGAGGAAGC TATGGTGCAC TCTCAGTACA
2451 ATCTGCTCTG ATGCCGCATA GTTAAGCCAG TATACACTCC GCTATCGCTA
2501 CGTGACTGGG TCATGGCTGC GCCCCGACAC CCGCCAACAC CCGCTGACGC
2551 GCCCTGACGG GCTTGTCTGC TCCCGGCATC CGCTTACAGA CAAGCTGTGA
2601 CCGTCTCCGG GAGCTGCATG TGTCAGAGGT TTTCACCGTC ATCACCGAAA
2651 CGCGCGAGGC AGCTGCGGTA AAGCTCATCA GCGTGGTCGT GCAGCGATTC
2701 ACAGATGTCT GCCTGTTCAT CCGCGTCCAG CTCGTTGAGT TTCTCCAGAA
2751 GCGTTAATGT CTGGCTTCTG ATAAAGCGGG CCATGTTAAG GGCGGTTTTT
2801 TCCTGTTTGG TCACTGATGC CTCCGTGTAA GGGGGATTTC TGTTCATGGG
2851 GGTAATGATA CCGATGAAAC GAGAGAGGAT GCTCACGATA CGGGTTACTG
2901 ATGATGAACA TGCCCGGTTA CTGGAACGTT GTGAGGGTAA ACAACTGGCG
2951 GTATGGATGC GGCGGGACCA GAGAAAAATC ACTCAGGGTC AATGCCAGCC
3001 GAACGCCAGC AAGACGTAGC CCAGCGCGTC GGCCGCCATG CCGGCGATAA
3051 TGGCCTGCTT CTCGCCGAAA CGTTTGGTGG CGGGACCAGT GACGAAGGCT
3101 TGAGCGAGGG CGTGCAAGAT TCCGAATACC GCAAGCGACA GGCCGATCAT
3151 CGTCGCGCTC CAGCGAAAGC GGTCCTCGCC GAAAATGACC CAGAGCGCTG
3201 CCGGCACCTG TCCTACGAGT TGCATGATAA AGAAGACAGT CATAAGTGCG
3251 GCGACGATAG TCATGCCCCG CGCCCACCGG AAGGAGCTGA CTGGGTTGAA
3301 GGCTCTCAAG GGCATCGGTC GAGATCCCGG TGCCTAATGA GTGAGCTAAC
3351 TTACATTAAT TGCGTTGCGC TCACTGCCCG CTTTCCAGTC GGGAAACCTG
3401 TCGTGCCAGC TGCATTAATG AATCGGCCAA CGCGCGGGGA GAGGCGGTTT
3451 GCGTATTGGG CGCCAGGGTG GTTTTTCTTT TCACCAGTGA GACGGGCAAC
3501 AGCTGATTGC CCTTCACCGC CTGGCCCTGA GAGAGTTGCA GCAAGCGGTC
3551 CACGCTGGTT TGCCCCAGCA GGCGAAAATC CTGTTTGATG GTGGTTAACG
3601 GCGGGATATA ACATGAGCTG TCTTCGGTAT CGTCGTATCC CACTACCGAG
3651 ATATCCGCAC CAACGCGCAG CCCGGACTCG GTAATGGCGC GCATTGCGCC
3701 CAGCGCCATC TGATCGTTGG CAACCAGCAT CGCAGTGGGA ACGATGCCCT
3751 CATTCAGCAT TTGCATGGTT TGTTGAAAAC CGGACATGGC ACTCCAGTCG
3801 CCTTCCCGTT CCGCTATCGG CTGAATTTGA TTGCGAGTGA GATATTTATG
3851 CCAGCCAGCC AGACGCAGAC GCGCCGAGAC AGAACTTAAT GGGCCCGCTA
3901 ACAGCGCGAT TTGCTGGTGA CCCAATGCGA CCAGATGCTC CACGCCCAGT
3951 CGCGTACCGT CTTCATGGGA GAAAATAATA CTGTTGATGG GTGTCTGGTC
4001 AGAGACATCA AGAAATAACG CCGGAACATT AGTGCAGGCA GCTTCCACAG
4051 CAATGGCATC CTGGTCATCC AGCGGATAGT TAATGATCAG CCCACTGACG
4101 CGTTGCGCGA GAAGATTGTG CACCGCCGCT TTACAGGCTT CGACGCCGCT
4151 TCGTTCTACC ATCGACACCA CCACGCTGGC ACCCAGTTGA TCGGCGCGAG
4201 ATTTAATCGC CGCGACAATT TGCGACGGCG CGTGCAGGGC CAGACTGGAG
4251 GTGGCAACGC CAATCAGCAA CGACTGTTTG CCCGCCAGTT GTTGTGCCAC
4301 GCGGTTGGGA ATGTAATTCA GCTCCGCCAT CGCCGCTTCC ACTTTTTCCC
4351 GCGTTTTCGC AGAAACGTGG CTGGCCTGGT TCACCACGCG GGAAACGGTC
4401 TGATAAGAGA CACCGGCATA CTCTGCGACA TCGTATAACG TTACTGGTTT
4451 CACATTCACC ACCCTGAATT GACTCTCTTC CGGGCGCTAT CATGCCATAC
4501 CGCGAAAGGT TTTGCGCCAT TCGATGGTGT CCGGGATCTC GACGCTCTCC
4551 CTTATGCGAC TCCTGCATTA GGAAGCAGCC CAGTAGTAGG TTGAGGCCGT
4601 TGAGCACCGC CGCCGCAAGG AATGGTGCAT GCAAGGAGAT GGCGCCCAAC
4651 AGTCCCCCGG CCACGGGGCC TGCCACCATA CCCACGCCGA AACAAGCGCT
4701 CATGAGCCCG AAGTGGCGAG CCCGATCTTC CCCATCGGTG ATGTCGGCGA
4751 TATAGGCGCC AGCAACCGCA CCTGTGGCGC CGGTGATGCC GGCCACGATG
4801 CGTCCGGCGT AGAGGATCGA GATCTCGATC CCGCGAAATT AATACGACTC
4851 ACTATAGGGG AATTGTGAGC GGATAACAAT TCCCCTCTAG AAATAATTTT
4901 GTTTAACTTT AAGAAGGAGA TATACTTATG AAAATCGAAG AAGGTAAACT
4951 GGTAATCGGT TCCCTGGAGG GTTGCTTTGC CAAGGGTACC AATGTTTTAA
5001 TGGCGGATGG GTCTATTGAA TGTATTGAAA ACATTGAGGT TGGTAATAAG
5051 GTCATGGGTA AAGATGGCAG ACCTCGTGAG GTAATTAAAT TGCCCAGAGG
5101 AAGAGAAACT ATGTACAGCG TCGTGCAGAA AAGTCAGCAC AGAGCCCACA
5151 AAAGTGACTC AAGTCGTGAA GTGCCAGAAT TACTCAAGTT TACGTGTAAT
5201 GCGACCCATG AGTTGGTTGT TAGAACACCT CGTAGTGTCC GCCGTTTGTC
5251 TCGTACCATT AAGGGTGTCG AATATTTTGA AGTTATTACT TTTGAGATGG
5301 GCCAAAAGAA AGCCCCCGAC GGTAGAATTG TTGAGCTTGT CAAGGAAGTT
5351 TCAAAGAGCT ACCCAATATC TGAGGGGCCT GAGAGAGCCA ACGAATTAGT
5401 AGAATCCTAT AGAAAGGCTT CAAATAAAGC TTATTTTGAG TGGACTATTG
5451 AGGCCAGAGA TCTTTCTCTG TTGGGTTCCC ATGTTCGTAA AGCTACCTAC
5501 CAGACTTACG CTCCAATTCT TTATGAGAAT GACCACTTTT TCGACTACAT
5551 GCAAAAAAGT AAGTTTCATC TCACCATTGA AGGTCCAAAA GTACTTGCTT
5601 ATTTACTTGG TTTATGGATT GGTGATGGAT TGTCTGACAG GGCAACTTTT
5651 TCGGTTGATT CCAGAGATAC TTCTTTGATG GAACGTGTTA CTGAATATGC
5701 TGAAAAGTTG AATTTGTGCG CCGAGTATAA GGACAGAAAA GAACCACAAG
5751 TTGCCAAAAC TGTTAATTTG TACTCTAAAG TTGTCAGAGG TGCTAGCACA
5801 AATCCTGGTG TATCCGCTTG GCAGGTCAAC ACAGCTTATA CTGCGGGACA
5851 ATTGGTCACA TATAACGGCA AGACGTATAA ATGTTTGCAG CCCCACACCT
5901 CCTTGGCAGG ATGGGAACCA TCCAACGTTC CTGCCTTGTG GCAGCTTCAA
5951 GGTGGCCATG GTGGTATTCG CAATAATCTT AATACTGAGA ATCCATTATG
6001 GGACGCTATT GTTGGCTTAG GATTCTTGAA GGACGGTGTC AAAAATATTC
6051 CTTCTTTCTT GTCTACGGAC AATATCGGTA CTCGTGAAAC ATTTCTTGCT
6101 GGTCTAATTG ATTCTGATGG CTATGTTACT GATGAGCATG GTATTAAAGC
6151 AACAATAAAG ACAATTCATA CTTCTGTCAG AGATGGTTTG GTTTCCCTTG
6201 CTCGTTCTTT AGGCTTAGTA GTCTCGGTTA ACGCAGAACC TGCTAAGGTT
6251 GACATGAATG TCACCAAACA TAAAATTAGT TATGCTATTT ATATGTCTGG
6301 TGGAGATGTT TTGCTTAACG TTCTTTCGAA GTGTGCCGGC TCTAAAAAAT
6351 TCAGGCCTGC TCCCGCCGCT GCTTTTGCAC GTGAGTGCCG CGGATTTTAT
6401 TTCGAGTTAC AAGAATTGAA GGAAGACGAT TATTATGGGA TTACTTTATC
6451 TGATGATTCT GATCATCAGT TTTTGCTTGG ATCCCAGGTT GTTGTACAGA
6501 ACAGAAGAGC TACTAGTTCG CGAGTCGACG GCGGCCGCGA ATTCCTCGAG
6551 CCCGGGTGAC TGCAGGAAGG GGATCCGGCT GCTAACAAAG CCCGAAAGGA
6601 AGCTGAGTTG GCTGCTGCCA CCGCTGAGCA ATAACTAGCA TAACCCCTTG
6651 GGGCCTCTAA ACGGGTCTTG AGGGGTTTTT TGCTGAAAGG AGGAACTATA
6701 TCCGGATATC CCGCAAGAGG CCCGGCAGTA CCGGCATAAC CAAGCCTATG
6751 CCTACAGCAT CCAGGGTGAC GGTGCCGAGG ATGACGATGA GCGCATTGTT
6801 AGATTTCATA CACGGTGCCT GACTGCGTTA GCAATTTAAC TGTGATAAAC
6851 TACCGCATTA AAGCTTGGCA CTGGCCGTCG TTTTACAACG TCGTGACTGG
6901 GAAAACCCTG GCGTTACCCA ACTTAATCGC CTTGCAGCAC ATCCCCCTTT
6951 CGCCAGCTGG CGTAATAGCG AAGAGGCCCG CACCGATCGC CCTTCCCAAC
7001 AGTTGCGCAG CCTGAATGGC GAATGGCAGC TTGGCTGTTT TGGCGGATGA
7051 GATAAGATTT TCAGCCTGAT ACAGATTAAA TCAGAACGCA GAAGCGGTCT
7101 GATAAAACAG AATTTGCCTG GCGGCAGTAG CGCGGTGGTC CCACCTGACC
7151 CCATGCCGAA CTCAGAAGTG AAACGCCGTA GCGCCGATGG TAGTGTGGGG
7201 TCTCCCCATG CGAGAGTAGG GAACTGCCAG GCATCAAATA AAACGAAAGG
7251 CTCAGTCGAA AGACTGGGCC TTTCGTTTTA TCTGTTGTTT GTCGGTGAAC
7301 GCTCTCCTGA GTAGGACAAA TCCGCCGGGA GCGGATTTGA ACGTTGCGAA
7351 GCAACGGCCC GGAGGGTGGC GGGCAGGACG CCCGCCATAA ACTGCCAGGC
7401 ATCAAATTAA GC
