LOCUS pMAL-p4X 6720 bp DNA circular 06-JUN-2014 DEFINITION Cloning vector pMAL-p4X, complete sequence. ACCESSION VERSION KEYWORDS . SOURCE Cloning vector pMAL-p4X ORGANISM Cloning vector pMAL-p4X other sequences; artificial sequences; vectors. REFERENCE 1 (bases 1 to 6720) 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) In press REFERENCE 2 (bases 1 to 6720) 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..6720 /organism="Cloning vector pMAL-p4X" /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..2991 /gene="malE expression cassette" /note="malE sequence 1528..2703; multiple cloning site region 2704..2809; lacZalpha sequence 2810..2991" /codon_start=1 /product="maltose binding protein (MBP) fusion" /translation="MKIKTGARILALSALTTMMFSASALAKIEEGKLVIWINGDKGYN GLAEVGKKFEKDTGIKVTVEHPDKLEEKFPQVAATGDGPDIIFWAHDRFGGYAQSGLL AEITPDKAFQDKLYPFTWDAVRYNGKLIAYPIAVEALSLIYNKDLLPNPPKTWEEIPA LDKELKAKGKSALMFNLQEPYFTWPLIAADGGYAFKYENGKYDIKDVGVDNAGAKAGL TFLVDLIKNKHMNADTDYSIAEAAFNKGETAMTINGPWAWSNIDTSKVNYGVTVLPTF KGQPSKPFVGVLSAGINAASPNKELAKEFLENYLLTDEGLEAVNKDKPLGAVALKSYE EELVKDPRIAATMENAQKGEIMPNIPQMSAFWYAVRTAVINAASGRQTVDEALKDAQT NSSSNNNNNNNNNNLGIEGRISEFGSSRVDLQASLALAVVLQRRDWENPGVTQLNRLA AHPPFASWRNSEEARTDRPSQQLRSLNGEWQLGCFGG" primer_bind 2662..2685 /note="malE primer sequence" misc_feature 2770..2807 /gene="malE expression cassette" /note="multiple cloning site (EcoRI-HindIII)" terminator 3179..3382 /note="rrnB T1T2 transcription terminators" gene 3492..4352 /gene="bla" CDS 3492..4352 /gene="bla" /note="ampR (confers resistance to ampicillin)" /codon_start=1 /product="beta-lactamase" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVE YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA EIGASLIKHW" sig_peptide 3492..3560 /gene="bla" /note="required for secretion to the periplasm; cleaved off to form the mature beta-lactamase protein" rep_origin complement(4394..4907) /note="M13 origin of replication (+ -)" rep_origin 5018..5606 /note="pMB1 origin of replication (clockwise) (RNAII -35 to RNA/DNA switch point)" gene complement(6036..6227) /gene="rop" CDS complement(6036..6227) /gene="rop" /exception="alternative start codon" /codon_start=1 /transl_table=11 /product="ROP protein" /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDH ADELYRSCLARFGDDGENL" BASE COUNT 1714 a 1692 c 1783 g 1531 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 gatcgaggga 2761 aggatttcag aattcggatc ctctagagtc gacctgcagg caagcttggc actggccgtc 2821 gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca 2881 catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa 2941 cagttgcgca gcctgaatgg cgaatggcag cttggctgtt ttggcggatg agataagatt 3001 ttcagcctga tacagattaa atcagaacgc agaagcggtc tgataaaaca gaatttgcct 3061 ggcggcagta gcgcggtggt cccacctgac cccatgccga actcagaagt gaaacgccgt 3121 agcgccgatg gtagtgtggg gtctccccat gcgagagtag ggaactgcca ggcatcaaat 3181 aaaacgaaag gctcagtcga aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa 3241 cgctctcctg agtaggacaa atccgccggg agcggatttg aacgttgcga agcaacggcc 3301 cggagggtgg cgggcaggac gcccgccata aactgccagg catcaaatta agcagaaggc 3361 catcctgacg gatggccttt ttgcgtttct acaaactctt ttgtttattt ttctaaatac 3421 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa 3481 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat 3541 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc 3601 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga 3661 gttttcgccc cgaagaacgt ttcccaatga tgagcacttt taaagttctg ctatgtggcg 3721 cggtattatc ccgtgttgac gccgggcaag agcaactcgg tcgccgcata cactattctc 3781 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag 3841 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc 3901 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg 3961 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg 4021 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac 4081 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac 4141 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg 4201 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg 4261 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg 4321 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac 4381 tttagattga tttaccccgg ttgataatca gaaaagcccc aaaaacagga agattgtata 4441 agcaaatatt taaattgtaa acgttaatat tttgttaaaa ttcgcgttaa atttttgtta 4501 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga 4561 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa 4621 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga 4681 accatcaccc aaatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc 4741 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga 4801 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg 4861 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtaaa aggatctagg 4921 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact 4981 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg 5041 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc 5101 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata 5161 ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta 5221 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc 5281 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg 5341 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac 5401 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg 5461 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt 5521 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct 5581 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg 5641 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata 5701 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca 5761 gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg gtattttctc cttacgcatc 5821 tgtgcggtat ttcacaccgc atatatggtg cactctcagt acaatctgct ctgatgccgc 5881 atagttaagc cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga 5941 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac 6001 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg 6061 aaacgcgcga ggcagctgcg gtaaagctca tcagcgtggt cgtgcagcga ttcacagatg 6121 tctgcctgtt catccgcgtc cagctcgttg agtttctcca gaagcgttaa tgtctggctt 6181 ctgataaagc gggccatgtt aagggcggtt ttttcctgtt tggtcactga tgcctccgtg 6241 taagggggat ttctgttcat gggggtaatg ataccgatga aacgagagag gatgctcacg 6301 atacgggtta ctgatgatga acatgcccgg ttactggaac gttgtgaggg taaacaactg 6361 gcggtatgga tgcggcggga ccagagaaaa atcactcagg gtcaatgcca gcgcttcgtt 6421 aatacagatg taggtgttcc acagggtagc cagcagcatc ctgcgatgca gatccggaac 6481 ataatggtgc agggcgctga cttccgcgtt tccagacttt acgaaacacg gaaaccgaag 6541 accattcatg ttgttgctca ggtcgcagac gttttgcagc agcagtcgct tcacgttcgc 6601 tcgcgtatcg gtgattcatt ctgctaacca gtaaggcaac cccgccagcc tagccgggtc 6661 ctcaacgaca ggagcacgat catgcgcacc cgtggccagg acccaacgct gcccgaaatt //