LOCUS pMAL-p 6220 bp DNA circular 06-JUN-2014 DEFINITION Cloning vector pMAL-p, complete sequence. ACCESSION VERSION KEYWORDS . SOURCE Cloning vector pMAL-p ORGANISM Cloning vector pMAL-p other sequences; artificial sequences; vectors. REFERENCE 1 (bases 1 to 6220) 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 6220) 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..6220 /organism="Cloning vector pMAL-p" /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..3382 /gene="malE expression cassette" promoter 1406..1433 /gene="malE expression cassette" /note="Ptac promoter (clockwise)" CDS 1528..2988 /gene="malE expression cassette" /exception="alternative start codon" /note="malE sequence 1528..2703; multiple cloning site region 2704..2805; lacZalpha sequence 2806..2987" /codon_start=1 /transl_table=11 /product="maltose binding protein (MBP) fusion" /translation="MKIKTGARILALSALTTMMFSASALAKIEEGKLVIWINGDKGYN GLAEVGKKFEKDTGIKVTVEHPDKLEEKFPQVAATGDGPDIIFWAHDRFGGYAQSGLL AEITPDKAFQDKLYPFTWDAVRYNGKLIAYPIAVEALSLIYNKDLLPNPPKTWEEIPA LDKELKAKGKSALMFNLQEPYFTWPLIAADGGYAFKYENGKYDIKDVGVDNAGAKAGL TFLVDLIKNKHMNADTDYSIAEAAFNKGETAMTINGPWAWSNIDTSKVNYGVTVLPTF KGQPSKPFVGVLSAGINAASPNKELAKEFLENYLLTDEGLEAVNKDKPLGAVALKSYE EELAKDPRIAATMENAQKGEIMPNIPQMSAFWYAVRTAVINAASGRQTVDEALKDAQT NSSSVPGRGSIEGRPEFSKPTLDGSSRVDLQASLALAVVLQRRDWENPGVTQLNRLAA HPPFASWRNSEEARTDRPSQQLRSLNGEWQLGCFGG" primer_bind 2662..2685 /note="malE primer sequence" misc_feature 2709..2804 /gene="malE expression cassette" /note="multiple cloning site (SacI-HindIII)" terminator 3176..3378 /gene="malE expression cassette" /note="rrnB T1T2 transcription terminators" gene 3489..4349 /gene="bla" CDS 3489..4349 /gene="bla" /note="ampR (confers resistance to ampicillin)" /codon_start=1 /product="beta-lactamase" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVE YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA EIGASLIKHW" sig_peptide 3489..3557 /gene="bla" /note="required for secretion to the periplasm; cleaved off to form the mature beta-lactamase protein" rep_origin 4520..5108 /note="pMB1 origin of replication (clockwise) (RNAII -35 to RNA/DNA switch point)" gene complement(5536..5727) /gene="rop" CDS complement(5536..5727) /gene="rop" /exception="alternative start codon" /codon_start=1 /transl_table=11 /product="ROP protein" /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDH ADELYRSCLARFGDDGENL" BASE COUNT 1542 a 1586 c 1658 g 1434 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 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 gcgaaagagt tcctcgaaaa ctatctgctg 2461 actgatgaag gtctggaagc ggttaataaa gacaaaccgc tgggtgccgt agcgctgaag 2521 tcttacgagg aagagttggc gaaagatcca cgtattgccg ccaccatgga aaacgcccag 2581 aaaggtgaaa tcatgccgaa catcccgcag atgtccgctt tctggtatgc cgtgcgtact 2641 gcggtgatca acgccgccag cggtcgtcag actgtcgatg aagccctgaa agacgcgcag 2701 actaattcga gctcggtacc cggccgggga tccatcgagg gtaggcctga attcagtaaa 2761 cctaccctcg atggatcctc tagagtcgac ctgcaggcaa gcttggcact ggccgtcgtt 2821 ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat 2881 ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag 2941 ttgcgcagcc tgaatggcga atggcagctt ggctgttttg gcggatgaga gaagattttc 3001 agcctgatac agattaaatc agaacgcaga agcggtctga taaaacagaa tttgcctggc 3061 ggcagtagcg cggtggtccc acctgacccc atgccgaact cagaagtgaa acgccgtagc 3121 gccgatggta gtgtggggtc tccccatgcg agagtaggga actgccaggc atcaaataaa 3181 acgaaaggct cagtcgaaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc 3241 tctcctgagt aggacaaatc cgccgggagc ggatttgaac gttgcgaagc aacggcccgg 3301 agggtggcgg gcaggacgcc cgccataaac tgccaggcat caaattaagc agaaggccat 3361 cctgacggat ggcctttttg cgtttctaca aactcttttg tttatttttc taaatacatt 3421 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa 3481 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt 3541 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt 3601 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt 3661 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg 3721 tattatcccg tgttgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga 3781 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa 3841 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga 3901 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa 3961 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca 4021 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta 4081 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac 4141 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc 4201 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag 4261 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga 4321 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt 4381 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata 4441 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag 4501 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa 4561 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt 4621 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc 4681 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa 4741 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa 4801 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc 4861 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa 4921 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa 4981 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg 5041 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc 5101 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg 5161 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg 5221 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg 5281 aagcggaaga gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc 5341 gcatatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatacac 5401 tccgctatcg ctacgtgact gggtcatggc tgcgccccga cacccgccaa cacccgctga 5461 cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc 5521 cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga ggcagctgcg 5581 gtaaagctca tcagcgtggt cgtgaagcga ttcacagatg tctgcctgtt catccgcgtc 5641 cagctcgttg agtttctcca gaagcgttaa tgtctggctt ctgataaagc gggccatgtt 5701 aagggcggtt ttttcctgtt tggtcactga tgcctccgtg taagggggat ttctgttcat 5761 gggggtaatg ataccgatga aacgagagag gatgctcacg atacgggtta ctgatgatga 5821 acatgcccgg ttactggaac gttgtgaggg taaacaactg gcggtatgga tgcggcggga 5881 ccagagaaaa atcactcagg gtcaatgcca gcgcttcgtt aatacagatg taggtgttcc 5941 acagggtagc cagcagcatc ctgcgatgca gatccggaac ataatggtgc agggcgctga 6001 cttccgcgtt tccagacttt acgaaacacg gaaaccgaag accattcatg ttgttgctca 6061 ggtcgcagac gttttgcagc agcagtcgct tcacgttcgc tcgcgtatcg gtgattcatt 6121 ctgctaacca gtaaggcaac cccgccagcc tagccgggtc ctcaacgaca ggagcacgat 6181 catgcgcacc cgtggccagg acccaacgct gcccgaaatt //