LOCUS pMYB5 8602 bp DNA circular 05-DEC-2007 DEFINITION Control vector pMYB5, complete sequence. ACCESSION VERSION KEYWORDS . SOURCE Control vector pMYB5 ORGANISM Control vector pMYB5 other sequences; artificial sequences; vectors. REFERENCE 1 (bases 1 to 8602) AUTHORS Chong,S., Mersha,F.B., Comb,D.G., Scott,M.E., Landry,D., Vence,L.M., Perler,F.B., Benner,J., Kucera,R.B., Hirvonen,C.A., Pelletier,J.J., Paulus,H. and Xu,M.-Q. TITLE Single-column purification of free recombinant proteins using a self-cleavable affinity tag derived from a protein splicing element JOURNAL Gene 192, 271-281 (1997) PUBMED 9224900 REFERENCE 2 (bases 1 to 8602) AUTHORS Perler,F.B. TITLE InBase, the intein database JOURNAL Nucleic Acids Res 30 (1), 383-384 (2002) PUBMED 11752343 REFERENCE 3 (bases 1 to 8602) AUTHORS Ghosh,I. and Xu,M.Q. TITLE Direct Submission JOURNAL Submitted (05-DEC-2007) Research Department, New England Biolabs, 240 County Road, Ipswich, MA 01938, USA COMMENT Derived from cloning vector pTYB1. FEATURES Location/Qualifiers source 1..8602 /organism="Control vector pMYB5" /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 5637..5656 /note="T7 universal primer sequence (NEB #S1248S)" promoter 5637..5654 /note="T7 promoter (transcript start 5654 clockwise)" protein_bind 5656..5680 /note="lac operator" /bound_moiety="LacI" RBS 5710..5716 /note="RBS for malE-intein-CBD CDS" gene 5725..8454 /gene="malE-intein-CBD CDS" CDS 5725..8454 /gene="malE-intein-CBD CDS" /note="malE sequence 5725..6825; multiple cloning site (MCS)/Factor Xa cleavage site (from pMAL-p2X) 6826..6885; Sce VMA intein sequence 6901..8262; chitin binding domain (CBD) sequence 8296..8454" /codon_start=1 /product="MBP/intein/CBD fusion" /translation="MKIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKL EEKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNG KLIAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWP LIAADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAF NKGETAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKEL AKEFLENYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNI PQMSAFWYAVRTAVINAASGRQTVDEALKDAQTNSSSNNNNNNNNNNLGIEGRGTLEG CFAKGTNVLMADGSIECIENIEVGNKVMGKDGRPREVIKLPRGRETMYSVVQKSQHRA HKSDSSREVPELLKFTCNATHELVVRTPRSVRRLSRTIKGVEYFEVITFEMGQKKAPD GRIVELVKEVSKSYPISEGPERANELVESYRKASNKAYFEWTIEARDLSLLGSHVRKA TYQTYAPILYENDHFFDYMQKSKFHLTIEGPKVLAYLLGLWIGDGLSDRATFSVDSRD TSLMERVTEYAEKLNLCAEYKDRKEPQVAKTVNLYSKVVRGNGIRNNLNTENPLWDAI VGLGFLKDGVKNIPSFLSTDNIGTRETFLAGLIDSDGYVTDEHGIKATIKTIHTSVRD GLVSLARSLGLVVSVNAEPAKVDMNVTKHKISYAIYMSGGDVLLNVLSKCAGSKKFRP APAAAFARECRGFYFELQELKEDDYYGITLSDDSDHQFLLGSQVVVHACGGLTGLNSG LTTNPGVSAWQVNTAYTAGQLVTYNGKTYKCLQPHTSLAGWEPSNVPALWQLQ" primer_bind complement(6964..6987) /note="intein reverse primer sequence (NEB #S1261S)" terminator 8474..8596 /note="T7 Tphi transcription terminator" BASE COUNT 2211 a 2074 c 2275 g 2042 t 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 acatatgaaa atcgaagaag gtaaactggt aatctggatt 5761 aacggcgata aaggctataa cggtctcgct gaagtcggta agaaattcga gaaagatacc 5821 ggaattaaag tcaccgttga gcatccggat aaactggaag agaaattccc acaggttgcg 5881 gcaactggcg atggccctga cattatcttc tgggcacacg accgctttgg tggctacgct 5941 caatctggcc tgttggctga aatcaccccg gacaaagcgt tccaggacaa gctgtatccg 6001 tttacctggg atgccgtacg ttacaacggc aagctgattg cttacccgat cgctgttgaa 6061 gcgttatcgc tgatttataa caaagatctg ctgccgaacc cgccaaaaac ctgggaagag 6121 atcccggcgc tggataaaga actgaaagcg aaaggtaaga gcgcgctgat gttcaacctg 6181 caagaaccgt acttcacctg gccgctgatt gctgctgacg ggggttatgc gttcaagtat 6241 gaaaacggca agtacgacat taaagacgtg ggcgtggata acgctggcgc gaaagcgggt 6301 ctgaccttcc tggttgacct gattaaaaac aaacacatga atgcagacac cgattactcc 6361 atcgcagaag ctgcctttaa taaaggcgaa acagcgatga ccatcaacgg cccgtgggca 6421 tggtccaaca tcgacaccag caaagtgaat tatggtgtaa cggtactgcc gaccttcaag 6481 ggtcaaccat ccaaaccgtt cgttggcgtg ctgagcgcag gtattaacgc cgccagtccg 6541 aacaaagagc tggcaaaaga gttcctcgaa aactatctgc tgactgatga aggtctggaa 6601 gcggttaata aagacaaacc gctgggtgcc gtagcgctga agtcttacga ggaagagttg 6661 gcgaaagatc cacgtattgc cgccactatg gaaaacgccc agaaaggtga aatcatgccg 6721 aacatcccgc agatgtccgc tttctggtat gccgtgcgta ctgcggtgat caacgccgcc 6781 agcggtcgtc agactgtcga tgaagccctg aaagacgcgc agactaattc gagctcgaac 6841 aacaacaaca ataacaataa caacaacctc gggatcgagg gaaggggtac gctcgagggg 6901 tgctttgcca agggtaccaa tgttttaatg gcggatgggt ctattgaatg tattgaaaac 6961 attgaggttg gtaataaggt catgggtaaa gatggcagac ctcgtgaggt aattaaattg 7021 cccagaggaa gagaaactat gtacagcgtc gtgcagaaaa gtcagcacag agcccacaaa 7081 agtgactcaa gtcgtgaagt gccagaatta ctcaagttta cgtgtaatgc gacccatgag 7141 ttggttgtta gaacacctcg tagtgtccgc cgtttgtctc gtaccattaa gggtgtcgaa 7201 tattttgaag ttattacttt tgagatgggc caaaagaaag cccccgacgg tagaattgtt 7261 gagcttgtca aggaagtttc aaagagctac ccaatatctg aggggcctga gagagccaac 7321 gaattagtag aatcctatag aaaggcttca aataaagctt attttgagtg gactattgag 7381 gccagagatc tttctctgtt gggttcccat gttcgtaaag ctacctacca gacttacgct 7441 ccaattcttt atgagaatga ccactttttc gactacatgc aaaaaagtaa gtttcatctc 7501 accattgaag gtccaaaagt acttgcttat ttacttggtt tatggattgg tgatggattg 7561 tctgacaggg caactttttc ggttgattcc agagatactt ctttgatgga acgtgttact 7621 gaatatgctg aaaagttgaa tttgtgcgcc gagtataagg acagaaaaga accacaagtt 7681 gccaaaactg ttaatttgta ctctaaagtt gtcagaggta atggtattcg caataatctt 7741 aatactgaga atccattatg ggacgctatt gttggcttag gattcttgaa ggacggtgtc 7801 aaaaatattc cttctttctt gtctacggac aatatcggta ctcgtgaaac atttcttgct 7861 ggtctaattg attctgatgg ctatgttact gatgagcatg gtattaaagc aacaataaag 7921 acaattcata cttctgtcag agatggtttg gtttcccttg ctcgttcttt aggcttagta 7981 gtctcggtta acgcagaacc tgctaaggtt gacatgaatg tcaccaaaca taaaattagt 8041 tatgctattt atatgtctgg tggagatgtt ttgcttaacg ttctttcgaa gtgtgccggc 8101 tctaaaaaat tcaggcctgc tcccgccgct gcttttgcac gtgagtgccg cggattttat 8161 ttcgagttac aagaattgaa ggaagacgat tattatggga ttactttatc tgatgattct 8221 gatcatcagt ttttgcttgg atcccaggtt gtcgtccatg catgcggtgg cctgaccggt 8281 ctgaactcag gcctcacgac aaatcctggt gtatccgctt ggcaggtcaa cacagcttat 8341 actgcgggac aattggtcac atataacggc aagacgtata aatgtttgca gccccacacc 8401 tccttggcag gatgggaacc atccaacgtt cctgccttgt ggcagcttca atgactgcag 8461 gaaggggatc cggctgctaa caaagcccga aaggaagctg agttggctgc tgccaccgct 8521 gagcaataac tagcataacc ccttggggcc tctaaacggg tcttgagggg ttttttgctg 8581 aaaggaggaa ctatatccgg at //