pTYB1
pTYB1
编号 | 载体名称 |
北京华越洋生物VECT5350 | pTYB1 |
pTYB1载体基本信息
载体名称: | pTYB1 |
质粒类型: | 大肠杆菌表达载体;蛋白纯化 |
高拷贝/低拷贝: | -- |
启动子: | T7 |
克隆方法: | 多克隆位点,限制性内切酶 |
载体大小: | 7477 bp |
5' 测序引物及序列: | T7: TAATACGACTCACTATAGGG |
3' 测序引物及序列: | -- |
载体标签: | Intein |
载体抗性: | Ampicillin |
筛选标记: | -- |
备注: | -- |
稳定性: | -- |
组成型: | -- |
病毒/非病毒: | 非病毒 |
pTYB1载体质粒图谱和多克隆位点信息
pTYB1载体简介
pTYB1 is an E. coli plasmid cloning vector designed for recombinant protein expression. It contains the pMB1 origin of replication from pBR322 and is maintained at a similar copy number to pBR322; in addition, pTYB1 also contains an M13 origin of replication. The multiple cloning site (MCS) is positioned to allow translational fusion of the Sce VMA intein tag to the C-terminus of the cloned target protein (1). The chitin binding domain (CBD) from B. circulans, fused to the C-terminus of the intein, facilitates purification of the intein-target protein precursor. Transcription of the gene fusion is controlled by the inducible T7 promoter, requiring E. coli strains containing integrated copies of the T7 RNA polymerase gene [e.g., C2566, C2833 or BL21(DE3)] for expression. Basal expression from the T7 promoter is minimized by the binding of the Lac repressor, encoded by the lacI gene, to the lac operator immediately downstream of the T7 promoter (3). Translation of the fusion utilizes the translation initiation signal (Shine Dalgarno sequence) from the strongly expressed T7 gene 10 protein (φ10). pTYB1, pTYB2, pTYB3, and pTYB4 are identical except for the MCS regions (opposite page). All four vectors contain either an NdeI (pTYB1 and pTYB2) or NcoI (pTYB3 and pTYB4) siteoverlapping the initiating methionine codon of the intein fusion gene. The N-terminal cysteine residue (“Cys1”) of the intein is shaded. Enzymes with unique restriction sites are shown in bold type. Location of sites of all NEB restriction enzymes can be found on the NEB web site (choose Technical Reference > DNA Sequences and Maps). Restriction site coordinates refer to the position of the 5′-most base on the top strand in each recognition sequence. Open reading frame (ORF) coordinates are in the form "translational start – translational stop”; numbers refer to positions on the top (clockwise) strand, regardless of the direction of transcription and include the start and stop codons. Component genes or regions of fusion ORFs are indented below the ORF itself. pMB1 origin of replication coordinates include the region from the -35 promoter sequence of the RNAII transcript to the RNA/DNA switch point. For the M13 origin, the arrow shows the direction of synthesis of the (+) strand, which gets packaged into phage particles. bla (ApR) gene coordinates include the signal sequence.
pTYB1载体序列
ORIGIN
1 AACTACGTCA GGTGGCACTT TTCGGGGAAA TGTGCGCGGA ACCCCTATTT GTTTATTTTT
61 CTAAATACAT TCAAATATGT ATCCGCTCAT GAGACAATAA CCCTGATAAA TGCTTCAATA
121 ATATTGAAAA AGGAAGAGTA TGAGTATTCA ACATTTCCGT GTCGCCCTTA TTCCCTTTTT
181 TGCGGCATTT TGCCTTCCTG TTTTTGCTCA CCCAGAAACG CTGGTGAAAG TAAAAGATGC
241 TGAAGATCAG TTGGGTGCAC GAGTGGGTTA CATCGAACTG GATCTCAACA GCGGTAAGAT
301 CCTTGAGAGT TTTCGCCCCG AAGAACGTTC TCCAATGATG AGCACTTTTA AAGTTCTGCT
361 ATGTGGCGCG GTATTATCCC GTGTTGACGC CGGGCAAGAG CAACTCGGTC GCCGCATACA
421 CTATTCTCAG AATGACTTGG TTGAGTACTC ACCAGTCACA GAAAAGCATC TTACGGATGG
481 CATGACAGTA AGAGAATTAT GCAGTGCTGC CATAACCATG AGTGATAACA CTGCGGCCAA
541 CTTACTTCTG ACAACGATCG GAGGACCGAA GGAGCTAACC GCTTTTTTGC ACAACATGGG
601 GGATCATGTA ACTCGCCTTG ATCGTTGGGA ACCGGAGCTG AATGAAGCCA TACCAAACGA
661 CGAGCGTGAC ACCACGATGC CTGTAGCAAT GGCAACAACG TTGCGCAAAC TATTAACTGG
721 CGAACTACTT ACTCTAGCTT CCCGGCAACA ATTAATAGAC TGGATGGAGG CGGATAAAGT
781 TGCAGGACCA CTTCTGCGCT CGGCCCTTCC GGCTGGCTGG TTTATTGCTG ATAAATCTGG
841 AGCCGGTGAG CGTGGGTCTC GCGGTATCAT TGCAGCACTG GGGCCAGATG GTAAGCCCTC
901 CCGTATCGTA GTTATCTACA CGACGGGGAG TCAGGCAACT ATGGATGAAC GAAATAGACA
961 GATCGCTGAG ATAGGTGCCT CACTGATTAA GCATTGGTAA CTGTCAGACC AAGTTTACTC
1021 ATATATACTT TAGATTGATT TACCCCGGTT GATAATCAGA AAAGCCCCAA AAACAGGAAG
1081 ATTGTATAAG CAAATATTTA AATTGTAAAC GTTAATATTT TGTTAAAATT CGCGTTAAAT
1141 TTTTGTTAAA TCAGCTCATT TTTTAACCAA TAGGCCGAAA TCGGCAAAAT CCCTTATAAA
1201 TCAAAAGAAT AGCCCGAGAT AGGGTTGAGT GTTGTTCCAG TTTGGAACAA GAGTCCACTA
1261 TTAAAGAACG TGGACTCCAA CGTCAAAGGG CGAAAAACCG TCTATCAGGG CGATGGCCCA
1321 CTACGTGAAC CATCACCCAA ATCAAGTTTT TTGGGGTCGA GGTGCCGTAA AGCACTAAAT
1381 CGGAACCCTA AAGGGAGCCC CCGATTTAGA GCTTGACGGG GAAAGCCGGC GAACGTGGCG
1441 AGAAAGGAAG GGAAGAAAGC GAAAGGAGCG GGCGCTAGGG CGCTGGCAAG TGTAGCGGTC
1501 ACGCTGCGCG TAACCACCAC ACCCGCCGCG CTTAATGCGC CGCTACAGGG CGCGTAAAAG
1561 GATCTAGGTG AAGATCCTTT TTGATAATCT CATGACCAAA ATCCCTTAAC GTGAGTTTTC
1621 GTTCCACTGA GCGTCAGACC CCGTAGAAAA GATCAAAGGA TCTTCTTGAG ATCCTTTTTT
1681 TCTGCGCGTA ATCTGCTGCT TGCAAACAAA AAAACCACCG CTACCAGCGG TGGTTTGTTT
1741 GCCGGATCAA GAGCTACCAA CTCTTTTTCC GAAGGTAACT GGCTTCAGCA GAGCGCAGAT
1801 ACCAAATACT GTCCTTCTAG TGTAGCCGTA GTTAGGCCAC CACTTCAAGA ACTCTGTAGC
1861 ACCGCCTACA TACCTCGCTC TGCTAATCCT GTTACCAGTG GCTGCTGCCA GTGGCGATAA
1921 GTCGTGTCTT ACCGGGTTGG ACTCAAGACG ATAGTTACCG GATAAGGCGC AGCGGTCGGG
1981 CTGAACGGGG GGTTCGTGCA CACAGCCCAG CTTGGAGCGA ACGACCTACA CCGAACTGAG
2041 ATACCTACAG CGTGAGCTAT GAGAAAGCGC CACGCTTCCC GAAGGGAGAA AGGCGGACAG
2101 GTATCCGGTA AGCGGCAGGG TCGGAACAGG AGAGCGCACG AGGGAGCTTC CAGGGGGAAA
2161 CGCCTGGTAT CTTTATAGTC CTGTCGGGTT TCGCCACCTC TGACTTGAGC GTCGATTTTT
2221 GTGATGCTCG TCAGGGGGGC GGAGCCTATG GAAAAACGCC AGCAACGCGG CCTTTTTACG
2281 GTTCCTGGCC TTTTGCTGGC CTTTTGCTCA CATGTTCTTT CCTGCGTTAT CCCCTGATTC
2341 TGTGGATAAC CGTATTACCG CCTTTGAGTG AGCTGATACC GCTCGCCGCA GCCGAACGAC
2401 CGAGCGCAGC GAGTCAGTGA GCGAGGAAGC TATGGTGCAC TCTCAGTACA ATCTGCTCTG
2461 ATGCCGCATA GTTAAGCCAG TATACACTCC GCTATCGCTA CGTGACTGGG TCATGGCTGC
2521 GCCCCGACAC CCGCCAACAC CCGCTGACGC GCCCTGACGG GCTTGTCTGC TCCCGGCATC
2581 CGCTTACAGA CAAGCTGTGA CCGTCTCCGG GAGCTGCATG TGTCAGAGGT TTTCACCGTC
2641 ATCACCGAAA CGCGCGAGGC AGCTGCGGTA AAGCTCATCA GCGTGGTCGT GCAGCGATTC
2701 ACAGATGTCT GCCTGTTCAT CCGCGTCCAG CTCGTTGAGT TTCTCCAGAA GCGTTAATGT
2761 CTGGCTTCTG ATAAAGCGGG CCATGTTAAG GGCGGTTTTT TCCTGTTTGG TCACTGATGC
2821 CTCCGTGTAA GGGGGATTTC TGTTCATGGG GGTAATGATA CCGATGAAAC GAGAGAGGAT
2881 GCTCACGATA CGGGTTACTG ATGATGAACA TGCCCGGTTA CTGGAACGTT GTGAGGGTAA
2941 ACAACTGGCG GTATGGATGC GGCGGGACCA GAGAAAAATC ACTCAGGGTC AATGCCAGCC
3001 GAACGCCAGC AAGACGTAGC CCAGCGCGTC GGCCGCCATG CCGGCGATAA TGGCCTGCTT
3061 CTCGCCGAAA CGTTTGGTGG CGGGACCAGT GACGAAGGCT TGAGCGAGGG CGTGCAAGAT
3121 TCCGAATACC GCAAGCGACA GGCCGATCAT CGTCGCGCTC CAGCGAAAGC GGTCCTCGCC
3181 GAAAATGACC CAGAGCGCTG CCGGCACCTG TCCTACGAGT TGCATGATAA AGAAGACAGT
3241 CATAAGTGCG GCGACGATAG TCATGCCCCG CGCCCACCGG AAGGAGCTGA CTGGGTTGAA
3301 GGCTCTCAAG GGCATCGGTC GAGATCCCGG TGCCTAATGA GTGAGCTAAC TTACATTAAT
3361 TGCGTTGCGC TCACTGCCCG CTTTCCAGTC GGGAAACCTG TCGTGCCAGC TGCATTAATG
3421 AATCGGCCAA CGCGCGGGGA GAGGCGGTTT GCGTATTGGG CGCCAGGGTG GTTTTTCTTT
3481 TCACCAGTGA GACGGGCAAC AGCTGATTGC CCTTCACCGC CTGGCCCTGA GAGAGTTGCA
3541 GCAAGCGGTC CACGCTGGTT TGCCCCAGCA GGCGAAAATC CTGTTTGATG GTGGTTAACG
3601 GCGGGATATA ACATGAGCTG TCTTCGGTAT CGTCGTATCC CACTACCGAG ATATCCGCAC
3661 CAACGCGCAG CCCGGACTCG GTAATGGCGC GCATTGCGCC CAGCGCCATC TGATCGTTGG
3721 CAACCAGCAT CGCAGTGGGA ACGATGCCCT CATTCAGCAT TTGCATGGTT TGTTGAAAAC
3781 CGGACATGGC ACTCCAGTCG CCTTCCCGTT CCGCTATCGG CTGAATTTGA TTGCGAGTGA
3841 GATATTTATG CCAGCCAGCC AGACGCAGAC GCGCCGAGAC AGAACTTAAT GGGCCCGCTA
3901 ACAGCGCGAT TTGCTGGTGA CCCAATGCGA CCAGATGCTC CACGCCCAGT CGCGTACCGT
3961 CTTCATGGGA GAAAATAATA CTGTTGATGG GTGTCTGGTC AGAGACATCA AGAAATAACG
4021 CCGGAACATT AGTGCAGGCA GCTTCCACAG CAATGGCATC CTGGTCATCC AGCGGATAGT
4081 TAATGATCAG CCCACTGACG CGTTGCGCGA GAAGATTGTG CACCGCCGCT TTACAGGCTT
4141 CGACGCCGCT TCGTTCTACC ATCGACACCA CCACGCTGGC ACCCAGTTGA TCGGCGCGAG
4201 ATTTAATCGC CGCGACAATT TGCGACGGCG CGTGCAGGGC CAGACTGGAG GTGGCAACGC
4261 CAATCAGCAA CGACTGTTTG CCCGCCAGTT GTTGTGCCAC GCGGTTGGGA ATGTAATTCA
4321 GCTCCGCCAT CGCCGCTTCC ACTTTTTCCC GCGTTTTCGC AGAAACGTGG CTGGCCTGGT
4381 TCACCACGCG GGAAACGGTC TGATAAGAGA CACCGGCATA CTCTGCGACA TCGTATAACG
4441 TTACTGGTTT CACATTCACC ACCCTGAATT GACTCTCTTC CGGGCGCTAT CATGCCATAC
4501 CGCGAAAGGT TTTGCGCCAT TCGATGGTGT CCGGGATCTC GACGCTCTCC CTTATGCGAC
4561 TCCTGCATTA GGAAGCAGCC CAGTAGTAGG TTGAGGCCGT TGAGCACCGC CGCCGCAAGG
4621 AATGGTGCAT GCCGGCATGC CGCCCTTTCG TCTTCAAGAA TTAATTCCCA ATTCCCCAGG
4681 CATCAAATAA AACGAAAGGC TCAGTCGAAA GACTGGGCCT TTCGTTTTAT CTGTTGTTTG
4741 TCGGTGAACG CTCTCCTGAG TAGGACAAAT CCGCCGGGAG CGGATTTGAA CGTTGCGAAG
4801 CAACGGCCCG GAGGGTGGCG GGCAGGACGC CCGCCATAAA CTGCCAGGAA TTAATTCCCC
4861 AGGCATCAAA TAAAACGAAA GGCTCAGTCG AAAGACTGGG CCTTTCGTTT TATCTGTTGT
4921 TTGTCGGTGA ACGCTCTCCT GAGTAGGACA AATCCGCCGG GAGCGGATTT GAACGTTGCG
4981 AAGCAACGGC CCGGAGGGTG GCGGGCAGGA CGCCCGCCAT AAACTGCCAG GAATTAATTC
5041 CCCAGGCATC AAATAAAACG AAAGGCTCAG TCGAAAGACT GGGCCTTTCG TTTTATCTGT
5101 TGTTTGTCGG TGAACGCTCT CCTGAGTAGG ACAAATCCGC CGGGAGCGGA TTTGAACGTT
5161 GCGAAGCAAC GGCCCGGAGG GTGGCGGGCA GGACGCCCGC CATAAACTGC CAGGAATTAA
5221 TTCCCCAGGC ATCAAATAAA ACGAAAGGCT CAGTCGAAAG ACTGGGCCTT TCGTTTTATC
5281 TGTTGTTTGT CGGTGAACGC TCTCCTGAGT AGGACAAATC CGCCGGGAGC GGATTTGAAC
5341 GTTGCGAAGC AACGGCCCGG AGGGTGGCGG GCAGGACGCC CGCCATAAAC TGCCAGGAAT
5401 TAATTCCCCA GGCATCAAAT AAAACGAAAG GCTCAGTCGA AAGACTGGGC CTTTCGTTTT
5461 ATCTGTTGTT TGTCGGTGAA CGCTCTCCTG AGTAGGACAA ATCCGCCGGG AGCGGATTTG
5521 AACGTTGCGA AGCAACGGCC CGGAGGGTGG CGGGCAGGAC GCCCGCCATA AACTGCCAGG
5581 AATTGGGGAT CGGAATTAAT TCCCGGTTTA AACCGGGGAT CTCGATCCCG CGAAATTAAT
5641 ACGACTCACT ATAGGGGAAT TGTGAGCGGA TAACAATTCC CCTCTAGAAA TAATTTTGTT
5701 TAACTTTAAG AAGGAGATAT ACATATGGCT AGCTCGCGAG TCGACGGCGG CCGCGAATTC
5761 CTCGAGGGCT CTTCCTGCTT TGCCAAGGGT ACCAATGTTT TAATGGCGGA TGGGTCTATT
5821 GAATGTATTG AAAACATTGA GGTTGGTAAT AAGGTCATGG GTAAAGATGG CAGACCTCGT
5881 GAGGTAATTA AATTGCCCAG AGGAAGAGAA ACTATGTACA GCGTCGTGCA GAAAAGTCAG
5941 CACAGAGCCC ACAAAAGTGA CTCAAGTCGT GAAGTGCCAG AATTACTCAA GTTTACGTGT
6001 AATGCGACCC ATGAGTTGGT TGTTAGAACA CCTCGTAGTG TCCGCCGTTT GTCTCGTACC
6061 ATTAAGGGTG TCGAATATTT TGAAGTTATT ACTTTTGAGA TGGGCCAAAA GAAAGCCCCC
6121 GACGGTAGAA TTGTTGAGCT TGTCAAGGAA GTTTCAAAGA GCTACCCAAT ATCTGAGGGG
6181 CCTGAGAGAG CCAACGAATT AGTAGAATCC TATAGAAAGG CTTCAAATAA AGCTTATTTT
6241 GAGTGGACTA TTGAGGCCAG AGATCTTTCT CTGTTGGGTT CCCATGTTCG TAAAGCTACC
6301 TACCAGACTT ACGCTCCAAT TCTTTATGAG AATGACCACT TTTTCGACTA CATGCAAAAA
6361 AGTAAGTTTC ATCTCACCAT TGAAGGTCCA AAAGTACTTG CTTATTTACT TGGTTTATGG
6421 ATTGGTGATG GATTGTCTGA CAGGGCAACT TTTTCGGTTG ATTCCAGAGA TACTTCTTTG
6481 ATGGAACGTG TTACTGAATA TGCTGAAAAG TTGAATTTGT GCGCCGAGTA TAAGGACAGA
6541 AAAGAACCAC AAGTTGCCAA AACTGTTAAT TTGTACTCTA AAGTTGTCAG AGGTAATGGT
6601 ATTCGCAATA ATCTTAATAC TGAGAATCCA TTATGGGACG CTATTGTTGG CTTAGGATTC
6661 TTGAAGGACG GTGTCAAAAA TATTCCTTCT TTCTTGTCTA CGGACAATAT CGGTACTCGT
6721 GAAACATTTC TTGCTGGTCT AATTGATTCT GATGGCTATG TTACTGATGA GCATGGTATT
6781 AAAGCAACAA TAAAGACAAT TCATACTTCT GTCAGAGATG GTTTGGTTTC CCTTGCTCGT
6841 TCTTTAGGCT TAGTAGTCTC GGTTAACGCA GAACCTGCTA AGGTTGACAT GAATGTCACC
6901 AAACATAAAA TTAGTTATGC TATTTATATG TCTGGTGGAG ATGTTTTGCT TAACGTTCTT
6961 TCGAAGTGTG CCGGCTCTAA AAAATTCAGG CCTGCTCCCG CCGCTGCTTT TGCACGTGAG
7021 TGCCGCGGAT TTTATTTCGA GTTACAAGAA TTGAAGGAAG ACGATTATTA TGGGATTACT
7081 TTATCTGATG ATTCTGATCA TCAGTTTTTG CTTGGATCCC AGGTTGTCGT CCATGCATGC
7141 GGTGGCCTGA CCGGTCTGAA CTCAGGCCTC ACGACAAATC CTGGTGTATC CGCTTGGCAG
7201 GTCAACACAG CTTATACTGC GGGACAATTG GTCACATATA ACGGCAAGAC GTATAAATGT
7261 TTGCAGCCCC ACACCTCCTT GGCAGGATGG GAACCATCCA ACGTTCCTGC CTTGTGGCAG
7321 CTTCAATGAC TGCAGGAAGG GGATCCGGCT GCTAACAAAG CCCGAAAGGA AGCTGAGTTG
7381 GCTGCTGCCA CCGCTGAGCA ATAACTAGCA TAACCCCTTG GGGCCTCTAA ACGGGTCTTG
7441 AGGGGTTTTT TGCTGAAAGG AGGAACTATA TCCGGAT
//
其他大肠杆菌表达载体: