LOCUS pMAL-p5X 5752 bp DNA circular 12-MAR-2014 DEFINITION Cloning vector pMAL-p5X, complete sequence. ACCESSION VERSION KEYWORDS . SOURCE Cloning vector pMAL-p5X ORGANISM Cloning vector pMAL-p5X other sequences; artificial sequences; vectors. REFERENCE 1 (bases 1 to 5752) 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 5752) AUTHORS Riggs,P.D. TITLE Direct Submission JOURNAL Submitted (10-NOV-2008) Research Department, New England Biolabs, 240 County Road, Ipswich, MA 01938, USA FEATURES Location/Qualifiers source 1..5752 /organism="Cloning vector pMAL-p5X" /mol_type="other DNA" gene 15..1163 /gene="lacIq" -35_signal 15..19 /gene="lacIq" /note="lacIq promoter (clockwise); TGGTG" -10_signal 40..46 /gene="lacIq" /note="lacIq promoter (clockwise); CATGATA" CDS 81..1163 /gene="lacIq" /codon_start=1 /transl_table=11 /product="lac repressor (LacI)" /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAE LNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERS GVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSII FSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREG DWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDT EDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNT QTASPRALADSLMQLARQVSRLESGQ" gene 1406..3043 /gene="malE expression cassette" -35_signal 1406..1411 /gene="malE expression cassette" /note="Ptac promoter (clockwise); TTGACA" -10_signal 1428..1433 /gene="malE expression cassette" /note="Ptac promoter (clockwise); TATAAT" CDS 1528..2829 /gene="malE expression cassette" /note="malE sequence 1528..2703; spacer and factor Xa site 2704..2762; multiple cloning site region 2763..2829" /codon_start=1 /product="maltose binding protein (MBP) fusion" /translation="MKIKTGARILALSALTTMMFSASALAKIEEGKLVIWINGDKGYN GLAEVGKKFEKDTGIKVTVEHPDKLEEKFPQVAATGDGPDIIFWAHDRFGGYAQSGLL AEITPDKAFQDKLYPFTWDAVRYNGKLIAYPIAVEALSLIYNKDLLPNPPKTWEEIPA LDKELKAKGKSALMFNLQEPYFTWPLIAADGGYAFKYENGKYDIKDVGVDNAGAKAGL TFLVDLIKNKHMNADTDYSIAEAAFNKGETAMTINGPWAWSNIDTSKVNYGVTVLPTF KGQPSKPFVGVLSAGINAASPNKELAKEFLENYLLTDEGLEAVNKDKPLGAVALKSYE EELVKDPRIAATMENAQKGEIMPNIPQMSAFWYAVRTAVINAASGRQTVDEALKDAQT NSSSNNNNNNNNNNLGIEGRISHMSMGGRDIVDGSEFPAGN" primer_bind 2662..2685 /note="malE primer sequence" misc_feature 2770..2822 /gene="malE expression cassette" /note="multiple cloning site (NdeI-SbfI)" terminator 2841..2884 /gene="malE expression cassette" /note="rrnB T1 transcription terminator" primer_bind complement(2898..2921) /note="MCS reverse primer sequence" terminator 3016..3043 /gene="malE expression cassette" /note="rrnB T2 transcription terminator" gene 3092..4022 /gene="bla" -35_signal 3092..3097 /gene="bla" /note="bla promoter (clockwise); TTCAAA" -10_signal 3115..3120 /gene="bla" /note="bla promoter (clockwise); GACAAT" CDS 3162..4022 /gene="bla" /note="ampR (confers resistance to ampicillin)" /codon_start=1 /product="beta-lactamase" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVE YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA EIGASLIKHW" rep_origin 4110..4698 /note="pMB1 origin of replication (clockwise) (RNAII -35 to RNA/DNA switch point)" gene complement(5068..5259) /gene="rop" CDS complement(5068..5259) /gene="rop" /codon_start=1 /transl_table=11 /product="ROP protein" /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDH ADELYRSCLARFGDDGENL" BASE COUNT 1438 a 1453 c 1537 g 1324 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 tatttcggta 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 gcaattgacc 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 gcaaaagagt tcctcgaaaa ctatctgctg 2461 actgatgaag gtctggaagc ggttaataaa gacaaaccgc tgggtgccgt agcgctgaag 2521 tcttacgagg aagagttggt gaaagatccg cgtattgccg ccactatgga aaacgcccag 2581 aaaggtgaaa tcatgccgaa catcccgcag atgtccgctt tctggtatgc cgtgcgtact 2641 gcggtgatca acgccgccag cggtcgtcag actgtcgatg aagccctgaa agacgcgcag 2701 actaattcga gctcgaacaa caacaacaat aacaataaca acaacctcgg gatcgaggga 2761 aggatttcac atatgtccat gggcggccgc gatatcgtcg acggatccga attccctgca 2821 ggtaattaaa taagcttcaa ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt 2881 ttatctgttg tttgtcggtg aacgctctcc tgagtaggac aaatccgccg ggagcggatt 2941 tgaacgttgc gaagcaacgg cccggagggt ggcgggcagg acgcccgcca taaactgcca 3001 ggcatcaaat taagcagaag gccatcctga cggatggcct ttttgcgttt ctacaaactc 3061 tttcggtccg ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat 3121 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc 3181 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa 3241 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac 3301 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt ttcccaatga 3361 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtgttgac gccgggcaag 3421 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca 3481 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca 3541 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa 3601 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc 3661 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa 3721 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag 3781 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct 3841 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac 3901 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa 3961 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt 4021 aactgtcaga ccaagtttac tcatatatac tttagattga tttccttagg actgagcgtc 4081 aaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc 4141 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta 4201 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt 4261 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc 4321 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg 4381 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg 4441 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag 4501 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc 4561 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat 4621 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg 4681 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc 4741 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt 4801 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca 4861 gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca tctgtgcggt 4921 atttcacacc gcatataagg tgcactgtga ctgggtcatg gctgcgcccc gacacccgcc 4981 aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc 5041 tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc 5101 gaggcagctg cggtaaagct catcagcgtg gtcgtgcagc gattcacaga tgtctgcctg 5161 ttcatccgcg tccagctcgt tgagtttctc cagaagcgtt aatgtctggc ttctgataaa 5221 gcgggccatg ttaagggcgg ttttttcctg tttggtcact gatgcctccg tgtaaggggg 5281 atttctgttc atgggggtaa tgataccgat gaaacgagag aggatgctca cgatacgggt 5341 tactgatgat gaacatgccc ggttactgga acgttgtgag ggtaaacaac tggcggtatg 5401 gatgcggcgg gaccagagaa aaatcactca gggtcaatgc cagcgcttcg ttaatacaga 5461 tgtaggtgtt ccacagggta gccagcagca tcctgcgatg cagatccgga acataatggt 5521 gcagggcgct gacttccgcg tttccagact ttacgaaaca cggaaaccga agaccattca 5581 tgttgttgct caggtcgcag acgttttgca gcagcagtcg cttcacgttc gctcgcgtat 5641 cggtgattca ttctgctaac cagtaaggca accccgccag cctagccggg tcctcaacga 5701 caggagcacg atcatgcgca cccgtggcca ggacccaacg ctgcccgaaa tt //