LOCUS pMAL-c2X 6645 bp DNA circular 06-JUN-2014 DEFINITION Cloning vector pMAL-c2X, complete sequence. ACCESSION VERSION KEYWORDS . SOURCE Cloning vector pMAL-c2X ORGANISM Cloning vector pMAL-c2X other sequences; artificial sequences; vectors. REFERENCE 1 (bases 1 to 6645) 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 6645) AUTHORS Riggs,P.D. TITLE Direct Submission JOURNAL Submitted (07-DEC-2007) Research Department, New England Biolabs, 240 County Road, Ipswich, MA 01938, USA FEATURES Location/Qualifiers source 1..6645 /organism="Cloning vector pMAL-c2X" /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..2916 /gene="malE expression cassette" /note="malE sequence 1528..2628; multiple cloning site region 2629..2734; lacZalpha sequence 2735..2916" /codon_start=1 /product="maltose binding protein (MBP) fusion" /translation="MKIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKL EEKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNG KLIAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWP LIAADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAF NKGETAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKEL AKEFLENYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNI PQMSAFWYAVRTAVINAASGRQTVDEALKDAQTNSSSNNNNNNNNNNLGIEGRISEFG SSRVDLQASLALAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRS LNGEWQLGCFGG" primer_bind 2587..2610 /note="malE primer sequence" misc_feature 2695..2732 /gene="malE expression cassette" /note="multiple cloning site (EcoRI-HindIII)" terminator 3104..3307 /gene="malE expression cassette" /note="rrnB T1T2 transcription terminators" gene 3417..4277 /gene="bla" CDS 3417..4277 /gene="bla" /note="ampR (confers resistance to ampicillin)" /codon_start=1 /product="beta-lactamase" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVE YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA EIGASLIKHW" sig_peptide 3417..3485 /gene="bla" /note="required for secretion to the periplasm; cleaved off to form the mature beta-lactamase protein" rep_origin complement(4319..4832) /note="M13 origin of replication (+ -)" rep_origin 4943..5531 /note="pMB1 origin of replication (clockwise) (RNAII -35 to RNA/DNA switch point)" gene complement(5961..6152) /gene="rop" CDS complement(5961..6152) /gene="rop" /exception="alternative start codon" /codon_start=1 /transl_table=11 /product="ROP protein" /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDH ADELYRSCLARFGDDGENL" BASE COUNT 1695 a 1670 c 1767 g 1513 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 aaaatcgaag 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 agctggcaaa agagttcctc gaaaactatc tgctgactga tgaaggtctg 2401 gaagcggtta ataaagacaa accgctgggt gccgtagcgc tgaagtctta cgaggaagag 2461 ttggcgaaag atccacgtat tgccgccact atggaaaacg cccagaaagg tgaaatcatg 2521 ccgaacatcc cgcagatgtc cgctttctgg tatgccgtgc gtactgcggt gatcaacgcc 2581 gccagcggtc gtcagactgt cgatgaagcc ctgaaagacg cgcagactaa ttcgagctcg 2641 aacaacaaca acaataacaa taacaacaac ctcgggatcg agggaaggat ttcagaattc 2701 ggatcctcta gagtcgacct gcaggcaagc ttggcactgg ccgtcgtttt acaacgtcgt 2761 gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc 2821 agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg 2881 aatggcgaat ggcagcttgg ctgttttggc ggatgagata agattttcag cctgatacag 2941 attaaatcag aacgcagaag cggtctgata aaacagaatt tgcctggcgg cagtagcgcg 3001 gtggtcccac ctgaccccat gccgaactca gaagtgaaac gccgtagcgc cgatggtagt 3061 gtggggtctc cccatgcgag agtagggaac tgccaggcat caaataaaac gaaaggctca 3121 gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc tcctgagtag 3181 gacaaatccg ccgggagcgg atttgaacgt tgcgaagcaa cggcccggag ggtggcgggc 3241 aggacgcccg ccataaactg ccaggcatca aattaagcag aaggccatcc tgacggatgg 3301 cctttttgcg tttctacaaa ctcttttgtt tatttttcta aatacattca aatatgtatc 3361 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga 3421 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt 3481 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag 3541 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag 3601 aacgtttccc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgtg 3661 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg 3721 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca 3781 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag 3841 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc 3901 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg 3961 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc 4021 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg 4081 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg 4141 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga 4201 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac 4261 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttac 4321 cccggttgat aatcagaaaa gccccaaaaa caggaagatt gtataagcaa atatttaaat 4381 tgtaaacgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt 4441 taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg 4501 gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt 4561 caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat cacccaaatc 4621 aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg 4681 atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa 4741 aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc 4801 cgccgcgctt aatgcgccgc tacagggcgc gtaaaaggat ctaggtgaag atcctttttg 4861 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg 4921 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc 4981 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc 5041 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt 5101 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc 5161 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact 5221 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac 5281 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag 5341 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg 5401 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg 5461 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga 5521 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt 5581 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct 5641 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg 5701 aggaagcgga agagcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac 5761 accgcatata tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta 5821 tacactccgc tatcgctacg tgactgggtc atggctgcgc cccgacaccc gccaacaccc 5881 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc 5941 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag 6001 ctgcggtaaa gctcatcagc gtggtcgtgc agcgattcac agatgtctgc ctgttcatcc 6061 gcgtccagct cgttgagttt ctccagaagc gttaatgtct ggcttctgat aaagcgggcc 6121 atgttaaggg cggttttttc ctgtttggtc actgatgcct ccgtgtaagg gggatttctg 6181 ttcatggggg taatgatacc gatgaaacga gagaggatgc tcacgatacg ggttactgat 6241 gatgaacatg cccggttact ggaacgttgt gagggtaaac aactggcggt atggatgcgg 6301 cgggaccaga gaaaaatcac tcagggtcaa tgccagcgct tcgttaatac agatgtaggt 6361 gttccacagg gtagccagca gcatcctgcg atgcagatcc ggaacataat ggtgcagggc 6421 gctgacttcc gcgtttccag actttacgaa acacggaaac cgaagaccat tcatgttgtt 6481 gctcaggtcg cagacgtttt gcagcagcag tcgcttcacg ttcgctcgcg tatcggtgat 6541 tcattctgct aaccagtaag gcaaccccgc cagcctagcc gggtcctcaa cgacaggagc 6601 acgatcatgc gcacccgtgg ccaggaccca acgctgcccg aaatt //