LOCUS pMAL-c5X-His 5695 bp DNA circular 12-MAR-2014 DEFINITION Cloning vector pMAL-c5X-His, complete sequence. ACCESSION VERSION KEYWORDS . SOURCE Cloning vector pMAL-c5X-His ORGANISM Cloning vector pMAL-c5X-His other sequences; artificial sequences; vectors. REFERENCE 1 (bases 1 to 5695) AUTHORS Riggs,P. TITLE Expression and purification of maltose-binding protein fusions JOURNAL (in) Asubel,F.A., Brent,R. and Kingston,R.E. (Eds.); CURRENT PROTOCOLS IN MOLECULAR BIOLOGY: 1-14; Greene Publishing and Wiley Interscience, New York, NY (1992) REFERENCE 2 (bases 1 to 5695) AUTHORS Riggs,P.D. TITLE Direct Submission JOURNAL Submitted (12-MAR-2014) Research Department, New England Biolabs, 240 County Road, Ipswich, MA 01938, USA FEATURES Location/Qualifiers source 1..5695 /organism="Cloning vector pMAL-c5X-His" /mol_type="other DNA" gene 15..1163 /gene="lacIq" -35_signal 15..19 /gene="lacIq" /note="lacIq promoter (clockwise); TGGTG" -10_signal 40..46 /gene="lacIq" /note="lacIq promoter (clockwise); CATGATA" CDS 81..1163 /gene="lacIq" /codon_start=1 /transl_table=11 /product="lac repressor (LacI)" /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAE LNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERS GVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSII FSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREG DWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDT EDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNT QTASPRALADSLMQLARQVSRLESGQ" gene 1406..2986 /gene="malE expression cassette" -35_signal 1406..1411 /gene="malE expression cassette" /note="Ptac promoter (clockwise); TTGACA" -10_signal 1428..1433 /gene="malE expression cassette" /note="Ptac promoter (clockwise); TATAAT" CDS 1528..2772 /gene="malE expression cassette" /note="malE sequence 1528..2628; spacer and factor Xa site 2629..2687; multiple cloning site region 2684..2747; 6x His tag 2752..2769" /codon_start=1 /product="maltose binding protein (MBP) fusion" /translation="MKIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKL EEKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNG KLIAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWP LIAADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAF NKGETAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKEL AKEFLENYLLTDEGLEAVNKDKPLGAVALKSYEEELVKDPRIAATMENAQKGEIMPNI PQMSAFWYAVRTAVINAASGRQTVDEALKDAQTNSSSNNNNNNNNNNLGIEGRISHMS MGGRDIVDGSEFPAGNHHHHHH" primer_bind 2587..2610 /note="malE primer sequence" misc_feature 2695..2747 /gene="malE expression cassette" /note="multiple cloning site (NdeI-SbfI)" misc_feature 2752..2769 /gene="malE expression cassette" /note="6x polyhistidine tag" terminator 2784..2827 /gene="malE expression cassette" /note="rrnB T1 transcription terminator" primer_bind complement(2841..2864) /note="MCS reverse primer sequence" terminator 2959..2986 /gene="malE expression cassette" /note="rrnB T2 transcription terminator" gene 3035..3965 /gene="bla" -35_signal 3035..3040 /gene="bla" /note="bla promoter (clockwise); TTCAAA" -10_signal 3058..3063 /gene="bla" /note="bla promoter (clockwise); GACAAT" CDS 3105..3965 /gene="bla" /note="ampR (confers resistance to ampicillin)" /codon_start=1 /product="beta-lactamase" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVE YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA EIGASLIKHW" rep_origin 4053..4641 /note="pMB1 origin of replication (clockwise) (RNAII -35 to RNA/DNA switch point)" gene complement(5011..5202) /gene="rop" CDS complement(5011..5202) /gene="rop" /codon_start=1 /transl_table=11 /product="ROP protein" /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDH ADELYRSCLARFGDDGENL" BASE COUNT 1424 a 1439 c 1523 g 1309 t ORIGIN 1 ccgacaccat cgaatggtgc aaaacctttc gcggtatggc atgatagcgc ccggaagaga 61 gtcaattcag ggtggtgaat gtgaaaccag taacgttata cgatgtcgca gagtatgccg 121 gtgtctctta tcagaccgtt tcccgcgtgg tgaaccaggc cagccacgtt tctgcgaaaa 181 cgcgggaaaa agtggaagcg gcgatggcgg agctgaatta cattcccaac cgcgtggcac 241 aacaactggc gggcaaacag tcgttgctga ttggcgttgc cacctccagt ctggccctgc 301 acgcgccgtc gcaaattgtc gcggcgatta aatctcgcgc cgatcaactg ggtgccagcg 361 tggtggtgtc gatggtagaa cgaagcggcg tcgaagcctg taaagcggcg gtgcacaatc 421 ttctcgcgca acgcgtcagt gggctgatca ttaactatcc gctggatgac caggatgcca 481 ttgctgtgga agctgcctgc actaatgttc cggcgttatt tcttgatgtc tctgaccaga 541 cacccatcaa cagtattatt ttctcccatg aagacggtac gcgactgggc gtggagcatc 601 tggtcgcatt gggtcaccag caaatcgcgc tgttagcggg cccattaagt tctgtctcgg 661 cgcgtctgcg tctggctggc tggcataaat atctcactcg caatcaaatt cagccgatag 721 cggaacggga aggcgactgg agtgccatgt ccggttttca acaaaccatg caaatgctga 781 atgagggcat cgttcccact gcgatgctgg ttgccaacga tcagatggcg ctgggcgcaa 841 tgcgcgccat taccgagtcc gggctgcgcg ttggtgcgga tatttcggta gtgggatacg 901 acgataccga agacagctca tgttatatcc cgccgttaac caccatcaaa caggattttc 961 gcctgctggg gcaaaccagc gtggaccgct tgctgcaact ctctcagggc caggcggtga 1021 agggcaatca gctgttgccc gtctcactgg tgaaaagaaa aaccaccctg gcgcccaata 1081 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt 1141 cccgactgga aagcgggcag tgagcgcaac gcaattaatg taagttagct cactcattag 1201 gcacaattct catgtttgac agcttatcat cgactgcacg gtgcaccaat gcttctggcg 1261 tcaggcagcc atcggaagct gtggtatggc tgtgcaggtc gtaaatcact gcataattcg 1321 tgtcgctcaa ggcgcactcc cgttctggat aatgtttttt gcgccgacat cataacggtt 1381 ctggcaaata ttctgaaatg agctgttgac aattaatcat cggctcgtat aatgtgtgga 1441 attgtgagcg gataacaatt tcacacagga aacagccagt ccgtttaggt gttttcacga 1501 gcaattgacc aacaaggacc atagattatg aaaatcgaag aaggtaaact ggtaatctgg 1561 attaacggcg ataaaggcta taacggtctc gctgaagtcg gtaagaaatt cgagaaagat 1621 accggaatta aagtcaccgt tgagcatccg gataaactgg aagagaaatt cccacaggtt 1681 gcggcaactg gcgatggccc tgacattatc ttctgggcac acgaccgctt tggtggctac 1741 gctcaatctg gcctgttggc tgaaatcacc ccggacaaag cgttccagga caagctgtat 1801 ccgtttacct gggatgccgt acgttacaac ggcaagctga ttgcttaccc gatcgctgtt 1861 gaagcgttat cgctgattta taacaaagat ctgctgccga acccgccaaa aacctgggaa 1921 gagatcccgg cgctggataa agaactgaaa gcgaaaggta agagcgcgct gatgttcaac 1981 ctgcaagaac cgtacttcac ctggccgctg attgctgctg acgggggtta tgcgttcaag 2041 tatgaaaacg gcaagtacga cattaaagac gtgggcgtgg ataacgctgg cgcgaaagcg 2101 ggtctgacct tcctggttga cctgattaaa aacaaacaca tgaatgcaga caccgattac 2161 tccatcgcag aagctgcctt taataaaggc gaaacagcga tgaccatcaa cggcccgtgg 2221 gcatggtcca acatcgacac cagcaaagtg aattatggtg taacggtact gccgaccttc 2281 aagggtcaac catccaaacc gttcgttggc gtgctgagcg caggtattaa cgccgccagt 2341 ccgaacaaag agctggcaaa agagttcctc gaaaactatc tgctgactga tgaaggtctg 2401 gaagcggtta ataaagacaa accgctgggt gccgtagcgc tgaagtctta cgaggaagag 2461 ttggtgaaag atccgcgtat tgccgccact atggaaaacg cccagaaagg tgaaatcatg 2521 ccgaacatcc cgcagatgtc cgctttctgg tatgccgtgc gtactgcggt gatcaacgcc 2581 gccagcggtc gtcagactgt cgatgaagcc ctgaaagacg cgcagactaa ttcgagctcg 2641 aacaacaaca acaataacaa taacaacaac ctcgggatcg agggaaggat ttcacatatg 2701 tccatgggcg gccgcgatat cgtcgacgga tccgaattcc ctgcaggtaa tcatcatcat 2761 caccaccact aaataagctt caaataaaac gaaaggctca gtcgaaagac tgggcctttc 2821 gttttatctg ttgtttgtcg gtgaacgctc tcctgagtag gacaaatccg ccgggagcgg 2881 atttgaacgt tgcgaagcaa cggcccggag ggtggcgggc aggacgcccg ccataaactg 2941 ccaggcatca aattaagcag aaggccatcc tgacggatgg cctttttgcg tttctacaaa 3001 ctctttcggt ccgttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac 3061 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt 3121 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag 3181 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg 3241 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgtttcccaa 3301 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtgtt gacgccgggc 3361 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag 3421 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa 3481 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc 3541 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg 3601 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa 3661 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa 3721 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg 3781 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag 3841 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg 3901 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt 3961 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttcctt aggactgagc 4021 gtcaaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc 4081 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag 4141 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc 4201 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac 4261 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc 4321 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt 4381 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt 4441 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc 4501 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt 4561 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca 4621 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt 4681 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt 4741 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag 4801 tcagtgagcg aggaagcgga agagcgcctg atgcggtatt ttctccttac gcatctgtgc 4861 ggtatttcac accgcatata aggtgcactg tgactgggtc atggctgcgc cccgacaccc 4921 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca 4981 agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg 5041 cgcgaggcag ctgcggtaaa gctcatcagc gtggtcgtgc agcgattcac agatgtctgc 5101 ctgttcatcc gcgtccagct cgttgagttt ctccagaagc gttaatgtct ggcttctgat 5161 aaagcgggcc atgttaaggg cggttttttc ctgtttggtc actgatgcct ccgtgtaagg 5221 gggatttctg ttcatggggg taatgatacc gatgaaacga gagaggatgc tcacgatacg 5281 ggttactgat gatgaacatg cccggttact ggaacgttgt gagggtaaac aactggcggt 5341 atggatgcgg cgggaccaga gaaaaatcac tcagggtcaa tgccagcgct tcgttaatac 5401 agatgtaggt gttccacagg gtagccagca gcatcctgcg atgcagatcc ggaacataat 5461 ggtgcagggc gctgacttcc gcgtttccag actttacgaa acacggaaac cgaagaccat 5521 tcatgttgtt gctcaggtcg cagacgtttt gcagcagcag tcgcttcacg ttcgctcgcg 5581 tatcggtgat tcattctgct aaccagtaag gcaaccccgc cagcctagcc gggtcctcaa 5641 cgacaggagc acgatcatgc gcacccgtgg ccaggaccca acgctgcccg aaatt //