LOCUS pMAL-c5X 5677 bp DNA circular 12-MAR-2014 DEFINITION Cloning vector pMAL-c5X, complete sequence. ACCESSION VERSION KEYWORDS . SOURCE Cloning vector pMAL-c5X ORGANISM Cloning vector pMAL-c5X other sequences; artificial sequences; vectors. REFERENCE 1 (bases 1 to 5677) 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 5677) AUTHORS Riggs,P.D. TITLE Direct Submission JOURNAL Submitted (13-NOV-2008) Research Department, New England Biolabs, 240 County Road, Ipswich, MA 01938, USA FEATURES Location/Qualifiers source 1..5677 /organism="Cloning vector pMAL-c5X" /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..2968 /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..2754 /gene="malE expression cassette" /note="malE sequence 1528..2628; spacer and factor Xa site 2629..2687; multiple cloning site region 2688..2754" /codon_start=1 /product="maltose binding protein (MBP) fusion" /translation="MKIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKL EEKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNG KLIAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWP LIAADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAF NKGETAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKEL AKEFLENYLLTDEGLEAVNKDKPLGAVALKSYEEELVKDPRIAATMENAQKGEIMPNI PQMSAFWYAVRTAVINAASGRQTVDEALKDAQTNSSSNNNNNNNNNNLGIEGRISHMS MGGRDIVDGSEFPAGN" primer_bind 2587..2610 /note="malE primer sequence" misc_feature 2695..2747 /gene="malE expression cassette" /note="multiple cloning site (NdeI-SbfI)" terminator 2766..2809 /gene="malE expression cassette" /note="rrnB T1 transcription terminator" primer_bind complement(2823..2846) /note="MCS reverse primer sequence" terminator 2941..2968 /gene="malE expression cassette" /note="rrnB T2 transcription terminator" gene 3017..3947 /gene="bla" -35_signal 3017..3022 /gene="bla" /note="bla promoter (clockwise); TTCAAA" -10_signal 3040..3045 /gene="bla" /note="bla promoter (clockwise); GACAAT" CDS 3087..3947 /gene="bla" /note="ampR (confers resistance to ampicillin)" /codon_start=1 /product="beta-lactamase" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVE YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA EIGASLIKHW" rep_origin 4035..4623 /note="pMB1 origin of replication (clockwise) (RNAII -35 to RNA/DNA switch point)" gene complement(4993..5184) /gene="rop" CDS complement(4993..5184) /gene="rop" /codon_start=1 /transl_table=11 /product="ROP protein" /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDH ADELYRSCLARFGDDGENL" BASE COUNT 1418 a 1430 c 1523 g 1306 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 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 ttggtgaaag atccgcgtat tgccgccact atggaaaacg cccagaaagg tgaaatcatg 2521 ccgaacatcc cgcagatgtc cgctttctgg tatgccgtgc gtactgcggt gatcaacgcc 2581 gccagcggtc gtcagactgt cgatgaagcc ctgaaagacg cgcagactaa ttcgagctcg 2641 aacaacaaca acaataacaa taacaacaac ctcgggatcg agggaaggat ttcacatatg 2701 tccatgggcg gccgcgatat cgtcgacgga tccgaattcc ctgcaggtaa ttaaataagc 2761 ttcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc tgttgtttgt 2821 cggtgaacgc tctcctgagt aggacaaatc cgccgggagc ggatttgaac gttgcgaagc 2881 aacggcccgg agggtggcgg gcaggacgcc cgccataaac tgccaggcat caaattaagc 2941 agaaggccat cctgacggat ggcctttttg cgtttctaca aactctttcg gtccgttgtt 3001 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc 3061 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc 3121 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa 3181 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg 3241 gtaagatcct tgagagtttt cgccccgaag aacgtttccc aatgatgagc acttttaaag 3301 ttctgctatg tggcgcggta ttatcccgtg ttgacgccgg gcaagagcaa ctcggtcgcc 3361 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta 3421 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg 3481 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca 3541 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac 3601 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat 3661 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg 3721 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata 3781 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta 3841 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa 3901 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag 3961 tttactcata tatactttag attgatttcc ttaggactga gcgtcaaccc cgtagaaaag 4021 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa 4081 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg 4141 aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag 4201 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg 4261 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga 4321 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc 4381 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc 4441 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga 4501 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt 4561 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg 4621 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac 4681 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga 4741 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg 4801 gaagagcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata 4861 taaggtgcac tgtgactggg tcatggctgc gccccgacac ccgccaacac ccgctgacgc 4921 gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg 4981 gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgaggc agctgcggta 5041 aagctcatca gcgtggtcgt gcagcgattc acagatgtct gcctgttcat ccgcgtccag 5101 ctcgttgagt ttctccagaa gcgttaatgt ctggcttctg ataaagcggg ccatgttaag 5161 ggcggttttt tcctgtttgg tcactgatgc ctccgtgtaa gggggatttc tgttcatggg 5221 ggtaatgata ccgatgaaac gagagaggat gctcacgata cgggttactg atgatgaaca 5281 tgcccggtta ctggaacgtt gtgagggtaa acaactggcg gtatggatgc ggcgggacca 5341 gagaaaaatc actcagggtc aatgccagcg cttcgttaat acagatgtag gtgttccaca 5401 gggtagccag cagcatcctg cgatgcagat ccggaacata atggtgcagg gcgctgactt 5461 ccgcgtttcc agactttacg aaacacggaa accgaagacc attcatgttg ttgctcaggt 5521 cgcagacgtt ttgcagcagc agtcgcttca cgttcgctcg cgtatcggtg attcattctg 5581 ctaaccagta aggcaacccc gccagcctag ccgggtcctc aacgacagga gcacgatcat 5641 gcgcacccgt ggccaggacc caacgctgcc cgaaatt //