LOCUS pMAL-c2G 6648 bp DNA circular 06-JUN-2014 DEFINITION Cloning vector pMAL-c2G, complete sequence. ACCESSION VERSION KEYWORDS . SOURCE Cloning vector pMAL-c2G ORGANISM Cloning vector pMAL-c2G other sequences; artificial sequences; vectors. REFERENCE 1 (bases 1 to 6648) AUTHORS Riggs,P. TITLE Expression and purification of maltose-binding protein fusions JOURNAL (in) Asubel,F.A., Brent,R., Kingston,R.E., Moore,D.D., Seidman,J.G., Smith,J.A. and Struhl,K. (Eds.); CURRENT PROTOCOLS IN MOLECULAR BIOLOGY: 1-14; Greene Publishing and Wiley Interscience, New York, NY (1992) REFERENCE 2 (bases 1 to 6648) AUTHORS Riggs,P.D. TITLE Direct Submission JOURNAL Submitted (07-DEC-2007) Research Department, New England Biolabs, 240 County Road, Ipswich, MA 01938, USA FEATURES Location/Qualifiers source 1..6648 /organism="Cloning vector pMAL-c2G" /mol_type="other DNA" gene 15..1163 /gene="lacIq" promoter 15..46 /gene="lacIq" /note="lacIq promoter (overexpressing mutant) (clockwise)" CDS 81..1163 /gene="lacIq" /exception="alternative start codon" /codon_start=1 /transl_table=11 /product="lac repressor (LacI)" /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAE LNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERS GVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSII FSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREG DWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDT EDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNT QTASPRALADSLMQLARQVSRLESGQ" gene 1406..3310 /gene="malE expression cassette" promoter 1406..1433 /gene="malE expression cassette" /note="Ptac promoter (clockwise)" CDS 1528..2919 /gene="malE expression cassette" /note="malE sequence 1528..2628; multiple cloning site region 2629..2737; lacZalpha sequence 2738..2919" /codon_start=1 /product="maltose binding protein (MBP) fusion" /translation="MKTEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKL EEKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNG KLIAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWP LIAADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAF NKGETAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKEL AKEFLENYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNI PQMSAFWYAVRTAVINAASGRQTVDEALKDAQTNSSSNNNNNNNNNNLGPGAAHYVEF GSSRVDLQASLALAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLR SLNGEWQLGCFGG" primer_bind 2587..2610 /note="malE primer sequence" misc_feature 2698..2735 /gene="malE expression cassette" /note="multiple cloning site (EcoRI-HindIII)" terminator 3107..3310 /gene="malE expression cassette" /note="rrnB T1T2 transcription terminators" gene 3420..4280 /gene="bla" CDS 3420..4280 /gene="bla" /note="ampR (confers resistance to ampicillin)" /codon_start=1 /product="beta-lactamase" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVE YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA EIGASLIKHW" sig_peptide 3420..3488 /gene="bla" /note="required for secretion to the periplasm; cleaved off to form the mature beta-lactamase protein" rep_origin complement(4322..4835) /note="M13 origin of replication (+ -)" rep_origin 4946..5534 /note="pMB1 origin of replication (clockwise) (RNAII -35 to RNA/DNA switch point)" gene complement(5964..6155) /gene="rop" CDS complement(5964..6155) /gene="rop" /exception="alternative start codon" /codon_start=1 /transl_table=11 /product="ROP protein" /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDH ADELYRSCLARFGDDGENL" BASE COUNT 1693 a 1675 c 1768 g 1512 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 tatctcggta 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 gcacttcacc aacaaggacc atagcatatg aaaactgaag 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 ttggcgaaag atccacgtat tgccgccact atggaaaacg cccagaaagg tgaaatcatg 2521 ccgaacatcc cgcagatgtc cgctttctgg tatgccgtgc gtactgcggt gatcaacgcc 2581 gccagcggtc gtcagactgt cgatgaagcc ctgaaagacg cgcagactaa ttcgagctcg 2641 aacaacaaca acaataacaa taacaacaac ctcgggccgg gtgcggcaca ctacgtagaa 2701 ttcggatcct ctagagtcga cctgcaggca agcttggcac tggccgtcgt tttacaacgt 2761 cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc 2821 gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc 2881 ctgaatggcg aatggcagct tggctgtttt ggcggatgag ataagatttt cagcctgata 2941 cagattaaat cagaacgcag aagcggtctg ataaaacaga atttgcctgg cggcagtagc 3001 gcggtggtcc cacctgaccc catgccgaac tcagaagtga aacgccgtag cgccgatggt 3061 agtgtggggt ctccccatgc gagagtaggg aactgccagg catcaaataa aacgaaaggc 3121 tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg tcggtgaacg ctctcctgag 3181 taggacaaat ccgccgggag cggatttgaa cgttgcgaag caacggcccg gagggtggcg 3241 ggcaggacgc ccgccataaa ctgccaggca tcaaattaag cagaaggcca tcctgacgga 3301 tggccttttt gcgtttctac aaactctttt gtttattttt ctaaatacat tcaaatatgt 3361 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta 3421 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg 3481 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac 3541 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg 3601 aagaacgttt cccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc 3661 gtgttgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg 3721 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat 3781 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg 3841 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg 3901 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc 3961 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt 4021 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct 4081 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc 4141 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca 4201 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct 4261 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt 4321 taccccggtt gataatcaga aaagccccaa aaacaggaag attgtataag caaatattta 4381 aattgtaaac gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt 4441 ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat 4501 agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa 4561 cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccaa 4621 atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc 4681 ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc 4741 gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac 4801 acccgccgcg cttaatgcgc cgctacaggg cgcgtaaaag gatctaggtg aagatccttt 4861 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc 4921 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct 4981 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa 5041 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag 5101 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc 5161 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg 5221 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca 5281 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat 5341 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg 5401 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc 5461 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc 5521 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc 5581 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg 5641 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga 5701 gcgaggaagc ggaagagcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt 5761 cacaccgcat atatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca 5821 gtatacactc cgctatcgct acgtgactgg gtcatggctg cgccccgaca cccgccaaca 5881 cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg 5941 accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg 6001 cagctgcggt aaagctcatc agcgtggtcg tgcagcgatt cacagatgtc tgcctgttca 6061 tccgcgtcca gctcgttgag tttctccaga agcgttaatg tctggcttct gataaagcgg 6121 gccatgttaa gggcggtttt ttcctgtttg gtcactgatg cctccgtgta agggggattt 6181 ctgttcatgg gggtaatgat accgatgaaa cgagagagga tgctcacgat acgggttact 6241 gatgatgaac atgcccggtt actggaacgt tgtgagggta aacaactggc ggtatggatg 6301 cggcgggacc agagaaaaat cactcagggt caatgccagc gcttcgttaa tacagatgta 6361 ggtgttccac agggtagcca gcagcatcct gcgatgcaga tccggaacat aatggtgcag 6421 ggcgctgact tccgcgtttc cagactttac gaaacacgga aaccgaagac cattcatgtt 6481 gttgctcagg tcgcagacgt tttgcagcag cagtcgcttc acgttcgctc gcgtatcggt 6541 gattcattct gctaaccagt aaggcaaccc cgccagccta gccgggtcct caacgacagg 6601 agcacgatca tgcgcacccg tggccaggac ccaacgctgc ccgaaatt //