LOCUS pMAL-c 6145 bp DNA circular 06-JUN-2014 DEFINITION Cloning vector pMAL-c, complete sequence. ACCESSION VERSION KEYWORDS . SOURCE Cloning vector pMAL-c ORGANISM Cloning vector pMAL-c other sequences; artificial sequences; vectors. REFERENCE 1 (bases 1 to 6145) 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 6145) AUTHORS Riggs,P. TITLE Direct Submission JOURNAL Submitted (30-APR-2008) Research Department, New England Biolabs, 240 County Road, Ipswich, MA 01938, USA FEATURES Location/Qualifiers source 1..6145 /organism="Cloning vector pMAL-c" /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..3307 /gene="malE expression cassette" promoter 1406..1433 /gene="malE expression cassette" /note="Ptac promoter (clockwise)" CDS 1528..2913 /gene="malE expression cassette" /note="malE sequence 1528..2628; multiple cloning site region 2629..2730; lacZalpha sequence 2731..2912" /codon_start=1 /product="maltose binding protein (MBP) fusion" /translation="MKTEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKL EEKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNG KLIAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWP LIAADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAF NKGETAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKEL AKEFLENYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNI PQMSAFWYAVRTAVINAASGRQTVDEALKDAQTNSSSVPGRGSIEGRPEFSKPTLDGS SRVDLQASLALAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSL NGEWQLGCFGG" primer_bind 2587..2610 /note="malE primer sequence" misc_feature 2634..2729 /gene="malE expression cassette" /note="multiple cloning site (SacI-HindIII)" terminator 3101..3303 /gene="malE expression cassette" /note="rrnB T1T2 transcription terminators" gene 3414..4274 /gene="bla" CDS 3414..4274 /gene="bla" /note="ampR (confers resistance to ampicillin)" /codon_start=1 /product="beta-lactamase" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVE YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA EIGASLIKHW" sig_peptide 3414..3482 /gene="bla" /note="required for secretion to the periplasm; cleaved off to form the mature beta-lactamase protein" rep_origin 4445..5033 /note="pMB1 origin of replication (clockwise) (RNAII -35 to RNA/DNA switch point)" gene complement(5461..5652) /gene="rop" CDS complement(5461..5652) /gene="rop" /exception="alternative start codon" /codon_start=1 /transl_table=11 /product="ROP protein" /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDH ADELYRSCLARFGDDGENL" BASE COUNT 1522 a 1563 c 1644 g 1416 t ORIGIN 1 ccgacaccat cgaatggcgc 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 atagattatg 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 agctggcgaa agagttcctc gaaaactatc tgctgactga tgaaggtctg 2401 gaagcggtta ataaagacaa accgctgggt gccgtagcgc tgaagtctta cgaggaagag 2461 ttggcgaaag atccacgtat tgccgccacc atggaaaacg cccagaaagg tgaaatcatg 2521 ccgaacatcc cgcagatgtc cgctttctgg tatgccgtgc gtactgcggt gatcaacgcc 2581 gccagcggtc gtcagactgt cgatgaagcc ctgaaagacg cgcagactaa ttcgagctcg 2641 gtacccggcc ggggatccat cgagggtagg cctgaattca gtaaacctac cctcgatgga 2701 tcctctagag tcgacctgca ggcaagcttg gcactggccg tcgttttaca acgtcgtgac 2761 tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc 2821 tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat 2881 ggcgaatggc agcttggctg ttttggcgga tgagagaaga ttttcagcct gatacagatt 2941 aaatcagaac gcagaagcgg tctgataaaa cagaatttgc ctggcggcag tagcgcggtg 3001 gtcccacctg accccatgcc gaactcagaa gtgaaacgcc gtagcgccga tggtagtgtg 3061 gggtctcccc atgcgagagt agggaactgc caggcatcaa ataaaacgaa aggctcagtc 3121 gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg aacgctctcc tgagtaggac 3181 aaatccgccg ggagcggatt tgaacgttgc gaagcaacgg cccggagggt ggcgggcagg 3241 acgcccgcca taaactgcca ggcatcaaat taagcagaag gccatcctga cggatggcct 3301 ttttgcgttt ctacaaactc ttttgtttat ttttctaaat acattcaaat atgtatccgc 3361 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta 3421 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg 3481 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg 3541 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac 3601 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtgttg 3661 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt 3721 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg 3781 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac 3841 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt 3901 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag 3961 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc 4021 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc 4081 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta 4141 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg 4201 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga 4261 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac 4321 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa 4381 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat 4441 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc 4501 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg 4561 gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc 4621 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg 4681 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg 4741 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa 4801 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg 4861 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga 4921 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct 4981 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca 5041 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc 5101 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg 5161 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc 5221 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata tggtgcactc 5281 tcagtacaat ctgctctgat gccgcatagt taagccagta tacactccgc tatcgctacg 5341 tgactgggtc atggctgcgc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc 5401 ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg 5461 tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag ctgcggtaaa gctcatcagc 5521 gtggtcgtga agcgattcac agatgtctgc ctgttcatcc gcgtccagct cgttgagttt 5581 ctccagaagc gttaatgtct ggcttctgat aaagcgggcc atgttaaggg cggttttttc 5641 ctgtttggtc actgatgcct ccgtgtaagg gggatttctg ttcatggggg taatgatacc 5701 gatgaaacga gagaggatgc tcacgatacg ggttactgat gatgaacatg cccggttact 5761 ggaacgttgt gagggtaaac aactggcggt atggatgcgg cgggaccaga gaaaaatcac 5821 tcagggtcaa tgccagcgct tcgttaatac agatgtaggt gttccacagg gtagccagca 5881 gcatcctgcg atgcagatcc ggaacataat ggtgcagggc gctgacttcc gcgtttccag 5941 actttacgaa acacggaaac cgaagaccat tcatgttgtt gctcaggtcg cagacgtttt 6001 gcagcagcag tcgcttcacg ttcgctcgcg tatcggtgat tcattctgct aaccagtaag 6061 gcaaccccgc cagcctagcc gggtcctcaa cgacaggagc acgatcatgc gcacccgtgg 6121 ccaggaccca acgctgcccg aaatt //