LOCUS pMAL-p4G 6723 bp DNA circular 06-JUN-2014 DEFINITION Cloning vector pMAL-p4G, complete sequence. ACCESSION VERSION KEYWORDS . SOURCE Cloning vector pMAL-p4G ORGANISM Cloning vector pMAL-p4G other sequences; artificial sequences; vectors. REFERENCE 1 (bases 1 to 6723) 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 6723) AUTHORS Riggs,P.D. TITLE Direct Submission JOURNAL Submitted (22-OCT-2007) Research Department, New England Biolabs, 240 County Road, Ipswich, MA 01938, USA FEATURES Location/Qualifiers source 1..6723 /organism="Cloning vector pMAL-p4G" /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..2994 /gene="malE expression cassette" /note="malE sequence 1528..2703; multiple cloning site region 2704..2812; lacZalpha sequence 2813..2994" /codon_start=1 /product="maltose binding protein (MBP) fusion" /translation="MKIKTGARILALSALTTMMFSASALAKIEEGKLVIWINGDKGYN GLAEVGKKFEKDTGIKVTVEHPDKLEEKFPQVAATGDGPDIIFWAHDRFGGYAQSGLL AEITPDKAFQDKLYPFTWDAVRYNGKLIAYPIAVEALSLIYNKDLLPNPPKTWEEIPA LDKELKAKGKSALMFNLQEPYFTWPLIAADGGYAFKYENGKYDIKDVGVDNAGAKAGL TFLVDLIKNKHMNADTDYSIAEAAFNKGETAMTINGPWAWSNIDTSKVNYGVTVLPTF KGQPSKPFVGVLSAGINAASPNKELAKEFLENYLLTDEGLEAVNKDKPLGAVALKSYE EELVKDPRIAATMENAQKGEIMPNIPQMSAFWYAVRTAVINAASGRQTVDEALKDAQT NSSSNNNNNNNNNNLGPGAAHYVEFGSSRVDLQASLALAVVLQRRDWENPGVTQLNRL AAHPPFASWRNSEEARTDRPSQQLRSLNGEWQLGCFGG" primer_bind 2662..2685 /note="malE primer sequence" misc_feature 2773..2810 /gene="malE expression cassette" /note="multiple cloning site (EcoRI-HindIII)" terminator 3182..3385 /note="rrnB T1T2 transcription terminators" gene 3495..4355 /gene="bla" CDS 3495..4355 /gene="bla" /note="ampR (confers resistance to ampicillin)" /codon_start=1 /product="beta-lactamase" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVE YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA EIGASLIKHW" sig_peptide 3495..3563 /gene="bla" /note="required for secretion to the periplasm; cleaved off to form the mature beta-lactamase protein" rep_origin complement(4397..4910) /note="M13 origin of replication (+ -)" rep_origin 5021..5609 /note="pMB1 origin of replication (clockwise) (RNAII -35 to RNA/DNA switch point)" gene complement(6039..6230) /gene="rop" CDS complement(6039..6230) /gene="rop" /exception="alternative start codon" /codon_start=1 /transl_table=11 /product="ROP protein" /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDH ADELYRSCLARFGDDGENL" BASE COUNT 1712 a 1697 c 1784 g 1530 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 aaaataaaaa caggtgcacg catcctcgca 1561 ttatccgcat taacgacgat gatgttttcc gcctcggctc tcgccaaaat cgaagaaggt 1621 aaactggtaa tctggattaa cggcgataaa ggctataacg gtctcgctga agtcggtaag 1681 aaattcgaga aagataccgg aattaaagtc accgttgagc atccggataa actggaagag 1741 aaattcccac aggttgcggc aactggcgat ggccctgaca ttatcttctg ggcacacgac 1801 cgctttggtg gctacgctca atctggcctg ttggctgaaa tcaccccgga caaagcgttc 1861 caggacaagc tgtatccgtt tacctgggat gccgtacgtt acaacggcaa gctgattgct 1921 tacccgatcg ctgttgaagc gttatcgctg atttataaca aagatctgct gccgaacccg 1981 ccaaaaacct gggaagagat cccggcgctg gataaagaac tgaaagcgaa aggtaagagc 2041 gcgctgatgt tcaacctgca agaaccgtac ttcacctggc cgctgattgc tgctgacggg 2101 ggttatgcgt tcaagtatga aaacggcaag tacgacatta aagacgtggg cgtggataac 2161 gctggcgcga aagcgggtct gaccttcctg gttgacctga ttaaaaacaa acacatgaat 2221 gcagacaccg attactccat cgcagaagct gcctttaata aaggcgaaac agcgatgacc 2281 atcaacggcc cgtgggcatg gtccaacatc gacaccagca aagtgaatta tggtgtaacg 2341 gtactgccga ccttcaaggg tcaaccatcc aaaccgttcg ttggcgtgct gagcgcaggt 2401 attaacgccg ccagtccgaa caaagagctg gcaaaagagt tcctcgaaaa ctatctgctg 2461 actgatgaag gtctggaagc ggttaataaa gacaaaccgc tgggtgccgt agcgctgaag 2521 tcttacgagg aagagttggt gaaagatccg cggattgccg ccactatgga aaacgcccag 2581 aaaggtgaaa tcatgccgaa catcccgcag atgtccgctt tctggtatgc cgtgcgtact 2641 gcggtgatca acgccgccag cggtcgtcag actgtcgatg aagccctgaa agacgcgcag 2701 actaattcga gctcgaacaa caacaacaat aacaataaca acaacctcgg gccgggtgcg 2761 gcacactacg tagaattcgg atcctctaga gtcgacctgc aggcaagctt ggcactggcc 2821 gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca 2881 gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc 2941 caacagttgc gcagcctgaa tggcgaatgg cagcttggct gttttggcgg atgagataag 3001 attttcagcc tgatacagat taaatcagaa cgcagaagcg gtctgataaa acagaatttg 3061 cctggcggca gtagcgcggt ggtcccacct gaccccatgc cgaactcaga agtgaaacgc 3121 cgtagcgccg atggtagtgt ggggtctccc catgcgagag tagggaactg ccaggcatca 3181 aataaaacga aaggctcagt cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt 3241 gaacgctctc ctgagtagga caaatccgcc gggagcggat ttgaacgttg cgaagcaacg 3301 gcccggaggg tggcgggcag gacgcccgcc ataaactgcc aggcatcaaa ttaagcagaa 3361 ggccatcctg acggatggcc tttttgcgtt tctacaaact cttttgttta tttttctaaa 3421 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt 3481 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg 3541 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag 3601 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg 3661 agagttttcg ccccgaagaa cgtttcccaa tgatgagcac ttttaaagtt ctgctatgtg 3721 gcgcggtatt atcccgtgtt gacgccgggc aagagcaact cggtcgccgc atacactatt 3781 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga 3841 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac 3901 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc 3961 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc 4021 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac 4081 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag 4141 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg 4201 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta 4261 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg 4321 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata 4381 tactttagat tgatttaccc cggttgataa tcagaaaagc cccaaaaaca ggaagattgt 4441 ataagcaaat atttaaattg taaacgttaa tattttgtta aaattcgcgt taaatttttg 4501 ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa 4561 agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa 4621 gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg 4681 tgaaccatca cccaaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa 4741 ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa 4801 ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct 4861 gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt aaaaggatct 4921 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc 4981 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc 5041 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg 5101 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa 5161 atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc 5221 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt 5281 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa 5341 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc 5401 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc 5461 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct 5521 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat 5581 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc 5641 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg 5701 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc 5761 gcagcgagtc agtgagcgag gaagcggaag agcgcctgat gcggtatttt ctccttacgc 5821 atctgtgcgg tatttcacac cgcatatatg gtgcactctc agtacaatct gctctgatgc 5881 cgcatagtta agccagtata cactccgcta tcgctacgtg actgggtcat ggctgcgccc 5941 cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct 6001 tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca 6061 ccgaaacgcg cgaggcagct gcggtaaagc tcatcagcgt ggtcgtgcag cgattcacag 6121 atgtctgcct gttcatccgc gtccagctcg ttgagtttct ccagaagcgt taatgtctgg 6181 cttctgataa agcgggccat gttaagggcg gttttttcct gtttggtcac tgatgcctcc 6241 gtgtaagggg gatttctgtt catgggggta atgataccga tgaaacgaga gaggatgctc 6301 acgatacggg ttactgatga tgaacatgcc cggttactgg aacgttgtga gggtaaacaa 6361 ctggcggtat ggatgcggcg ggaccagaga aaaatcactc agggtcaatg ccagcgcttc 6421 gttaatacag atgtaggtgt tccacagggt agccagcagc atcctgcgat gcagatccgg 6481 aacataatgg tgcagggcgc tgacttccgc gtttccagac tttacgaaac acggaaaccg 6541 aagaccattc atgttgttgc tcaggtcgca gacgttttgc agcagcagtc gcttcacgtt 6601 cgctcgcgta tcggtgattc attctgctaa ccagtaaggc aaccccgcca gcctagccgg 6661 gtcctcaacg acaggagcac gatcatgcgc acccgtggcc aggacccaac gctgcccgaa 6721 att //