LOCUS pTYB11 7412 bp DNA circular 23-OCT-2007 DEFINITION Cloning vector pTYB11, complete sequence. ACCESSION VERSION KEYWORDS . SOURCE Cloning vector pTYB11 ORGANISM Cloning vector pTYB11 other sequences; artificial sequences; vectors. REFERENCE 1 (bases 1 to 7412) AUTHORS Chong,S., Williams,K.S., Wotkowicz,C. and Xu,M.-Q. TITLE Modulation of protein splicing of the Saccharomyces cerevisiae vacuolar membrane ATPase intein JOURNAL J Biol Chem 273, 10567-10577 (1998) PUBMED 9553117 REFERENCE 2 (bases 1 to 7412) AUTHORS Chong,S., Montello,G.E., Zhang,A., Cantor,E.J., Liao,W., Xu,M.-Q. and Benner,J. TITLE Utilizing the C-terminal cleavage activity of a protein splicing element to purify recombinant proteins in a single chromatographic step JOURNAL Nucleic Acids Res 26, 5109-5115 (1998) PUBMED 9801307 REFERENCE 3 (bases 1 to 7412) AUTHORS Perler,F.B. TITLE InBase, the intein database JOURNAL Nucleic Acids Res 30 (1), 383-384 (2002) PUBMED 11752343 REFERENCE 4 (bases 1 to 7412) AUTHORS Ghosh,I. and Xu,M.Q. TITLE Direct Submission JOURNAL Submitted (23-OCT-2007) Research Department, New England Biolabs, 240 County Road, Ipswich, MA 01938, USA FEATURES Location/Qualifiers source 1..7412 /organism="Cloning vector pTYB11" /mol_type="other DNA" gene 140..1000 /gene="bla" CDS 140..1000 /gene="bla" /note="ampR (confers resistance to ampicillin)" /codon_start=1 /product="beta-lactamase" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY IELDLNSGKILESFRPEERSPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVE YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA EIGASLIKHW" sig_peptide 140..208 /gene="bla" /note="required for secretion to the periplasm; cleaved off to form the mature beta-lactamase protein" rep_origin 1042..1555 /note="M13 origin of replication (+ -)" rep_origin 1666..2254 /note="pMB1 origin of replication (clockwise) (RNAII -35 to RNA/DNA switch point)" gene complement(2623..2814) /gene="rop" CDS complement(2623..2814) /gene="rop" /exception="alternative start codon" /codon_start=1 /transl_table=11 /product="ROP protein" /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDH ADELYRSCLARFGDDGENL" gene complement(3371..4453) /gene="lacI" CDS complement(3371..4453) /gene="lacI" /exception="alternative start codon" /codon_start=1 /transl_table=11 /product="lac repressor (LacI)" /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAE LNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERS GVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSII FSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREG DWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDT EDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNT QTASPRALADSLMQLARQVSRLESGQ" primer_bind 4840..4859 /note="T7 universal primer sequence (NEB #S1248S)" promoter 4840..4857 /note="T7 promoter (transcript start 5654 clockwise)" protein_bind 4859..4883 /note="lac operator" /bound_moiety="LacI" RBS 4913..4919 /note="RBS for expression CDS" gene 4928..6559 /gene="expression CDS" CDS 4928..6559 /gene="expression CDS" /note="malE fragment (translational start) 4928..4972; Sce VMA intein 5' region 4973..5791; chitin binding domain (CBD) 5798..5950; Sce VMA intein 3' region 5963..6502; multiple cloning site (SapI-SmaI) 6503..6556" /codon_start=1 /product="intein/CBD fusion" /translation="MKIEEGKLVIGSLEGCFAKGTNVLMADGSIECIENIEVGNKVMG KDGRPREVIKLPRGRETMYSVVQKSQHRAHKSDSSREVPELLKFTCNATHELVVRTPR SVRRLSRTIKGVEYFEVITFEMGQKKAPDGRIVELVKEVSKSYPISEGPERANELVES YRKASNKAYFEWTIEARDLSLLGSHVRKATYQTYAPILYENDHFFDYMQKSKFHLTIE GPKVLAYLLGLWIGDGLSDRATFSVDSRDTSLMERVTEYAEKLNLCAEYKDRKEPQVA KTVNLYSKVVRGASTNPGVSAWQVNTAYTAGQLVTYNGKTYKCLQPHTSLAGWEPSNV PALWQLQGGHGGIRNNLNTENPLWDAIVGLGFLKDGVKNIPSFLSTDNIGTRETFLAG LIDSDGYVTDEHGIKATIKTIHTSVRDGLVSLARSLGLVVSVNAEPAKVDMNVTKHKI SYAIYMSGGDVLLNVLSKCAGSKKFRPAPAAAFARECRGFYFELQELKEDDYYGITLS DDSDHQFLLGSQVVVQNRRATSSRVDGGREFLEPG" primer_bind 6362..6385 /note="intein forward primer sequence (NEB #S1263S)" misc_feature 6503..6556 /gene="expression CDS" /note="multiple cloning site (NdeI-SapI)" terminator 6579..6701 /note="T7 Tphi transcription terminator" BASE COUNT 1859 a 1809 c 1954 g 1790 t ORIGIN 1 agaaggccat cctgacggat ggcctttttg cgtttctaca aactcttttt 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 gcaaggagat ggcgcccaac agtcccccgg ccacggggcc tgccaccata 4681 cccacgccga aacaagcgct catgagcccg aagtggcgag cccgatcttc cccatcggtg 4741 atgtcggcga tataggcgcc agcaaccgca cctgtggcgc cggtgatgcc ggccacgatg 4801 cgtccggcgt agaggatcga gatctcgatc ccgcgaaatt aatacgactc actatagggg 4861 aattgtgagc ggataacaat tcccctctag aaataatttt gtttaacttt aagaaggaga 4921 tatacttatg aaaatcgaag aaggtaaact ggtaatcggt tccctggagg gttgctttgc 4981 caagggtacc aatgttttaa tggcggatgg gtctattgaa tgtattgaaa acattgaggt 5041 tggtaataag gtcatgggta aagatggcag acctcgtgag gtaattaaat tgcccagagg 5101 aagagaaact atgtacagcg tcgtgcagaa aagtcagcac agagcccaca aaagtgactc 5161 aagtcgtgaa gtgccagaat tactcaagtt tacgtgtaat gcgacccatg agttggttgt 5221 tagaacacct cgtagtgtcc gccgtttgtc tcgtaccatt aagggtgtcg aatattttga 5281 agttattact tttgagatgg gccaaaagaa agcccccgac ggtagaattg ttgagcttgt 5341 caaggaagtt tcaaagagct acccaatatc tgaggggcct gagagagcca acgaattagt 5401 agaatcctat agaaaggctt caaataaagc ttattttgag tggactattg aggccagaga 5461 tctttctctg ttgggttccc atgttcgtaa agctacctac cagacttacg ctccaattct 5521 ttatgagaat gaccactttt tcgactacat gcaaaaaagt aagtttcatc tcaccattga 5581 aggtccaaaa gtacttgctt atttacttgg tttatggatt ggtgatggat tgtctgacag 5641 ggcaactttt tcggttgatt ccagagatac ttctttgatg gaacgtgtta ctgaatatgc 5701 tgaaaagttg aatttgtgcg ccgagtataa ggacagaaaa gaaccacaag ttgccaaaac 5761 tgttaatttg tactctaaag ttgtcagagg tgctagcaca aatcctggtg tatccgcttg 5821 gcaggtcaac acagcttata ctgcgggaca attggtcaca tataacggca agacgtataa 5881 atgtttgcag ccccacacct ccttggcagg atgggaacca tccaacgttc ctgccttgtg 5941 gcagcttcaa ggtggccatg gtggtattcg caataatctt aatactgaga atccattatg 6001 ggacgctatt gttggcttag gattcttgaa ggacggtgtc aaaaatattc cttctttctt 6061 gtctacggac aatatcggta ctcgtgaaac atttcttgct ggtctaattg attctgatgg 6121 ctatgttact gatgagcatg gtattaaagc aacaataaag acaattcata cttctgtcag 6181 agatggtttg gtttcccttg ctcgttcttt aggcttagta gtctcggtta acgcagaacc 6241 tgctaaggtt gacatgaatg tcaccaaaca taaaattagt tatgctattt atatgtctgg 6301 tggagatgtt ttgcttaacg ttctttcgaa gtgtgccggc tctaaaaaat tcaggcctgc 6361 tcccgccgct gcttttgcac gtgagtgccg cggattttat ttcgagttac aagaattgaa 6421 ggaagacgat tattatggga ttactttatc tgatgattct gatcatcagt ttttgcttgg 6481 atcccaggtt gttgtacaga acagaagagc tactagttcg cgagtcgacg gcggccgcga 6541 attcctcgag cccgggtgac tgcaggaagg ggatccggct gctaacaaag cccgaaagga 6601 agctgagttg gctgctgcca ccgctgagca ataactagca taaccccttg gggcctctaa 6661 acgggtcttg aggggttttt tgctgaaagg aggaactata tccggatatc ccgcaagagg 6721 cccggcagta ccggcataac caagcctatg cctacagcat ccagggtgac ggtgccgagg 6781 atgacgatga gcgcattgtt agatttcata cacggtgcct gactgcgtta gcaatttaac 6841 tgtgataaac taccgcatta aagcttggca ctggccgtcg ttttacaacg tcgtgactgg 6901 gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg 6961 cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc 7021 gaatggcagc ttggctgttt tggcggatga gataagattt tcagcctgat acagattaaa 7081 tcagaacgca gaagcggtct gataaaacag aatttgcctg gcggcagtag cgcggtggtc 7141 ccacctgacc ccatgccgaa ctcagaagtg aaacgccgta gcgccgatgg tagtgtgggg 7201 tctccccatg cgagagtagg gaactgccag gcatcaaata aaacgaaagg ctcagtcgaa 7261 agactgggcc tttcgtttta tctgttgttt gtcggtgaac gctctcctga gtaggacaaa 7321 tccgccggga gcggatttga acgttgcgaa gcaacggccc ggagggtggc gggcaggacg 7381 cccgccataa actgccaggc atcaaattaa gc //