LOCUS pMAL-c5G 5680 bp DNA circular 12-MAR-2014 DEFINITION Cloning vector pMAL-c5G, complete sequence. ACCESSION VERSION KEYWORDS . SOURCE Cloning vector pMAL-c5G ORGANISM Cloning vector pMAL-c5G other sequences; artificial sequences; vectors. REFERENCE 1 (bases 1 to 5680) 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 5680) 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..5680 /organism="Cloning vector pMAL-c5G" /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..2971 /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..2757 /gene="malE expression cassette" /note="malE sequence 1528..2628; spacer and Genenase I site 2629..2690; multiple cloning site region 2691..2757" /codon_start=1 /product="maltose binding protein (MBP) fusion" /translation="MKIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKL EEKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNG KLIAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWP LIAADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAF NKGETAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKEL AKEFLENYLLTDEGLEAVNKDKPLGAVALKSYEEELVKDPRIAATMENAQKGEIMPNI PQMSAFWYAVRTAVINAASGRQTVDEALKDAQTNSSSNNNNNNNNNNLGPGAAHYVHM SMGGRDIVDGSEFPAGN" primer_bind 2587..2610 /note="malE primer sequence" misc_feature 2692..2750 /gene="malE expression cassette" /note="multiple cloning site (SnaBI-SbfI)" terminator 2769..2812 /gene="malE expression cassette" /note="rrnB T1 transcription terminator" primer_bind complement(2826..2849) /note="MCS reverse primer sequence" terminator 2944..2971 /gene="malE expression cassette" /note="rrnB T2 transcription terminator" gene 3020..3950 /gene="bla" -35_signal 3020..3025 /gene="bla" /note="bla promoter (clockwise); TTCAAA" -10_signal 3043..3048 /gene="bla" /note="bla promoter (clockwise); GACAAT" CDS 3090..3950 /gene="bla" /note="ampR (confers resistance to ampicillin)" /codon_start=1 /product="beta-lactamase" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVE YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA EIGASLIKHW" rep_origin 4038..4626 /note="pMB1 origin of replication (clockwise) (RNAII -35 to RNA/DNA switch point)" gene complement(4996..5187) /gene="rop" CDS complement(4996..5187) /gene="rop" /codon_start=1 /transl_table=11 /product="ROP protein" /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDH ADELYRSCLARFGDDGENL" BASE COUNT 1416 a 1435 c 1524 g 1305 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 ctcgggccgg gtgcggcaca ctacgtacat 2701 atgtccatgg gcggccgcga tatcgtcgac ggatccgaat tccctgcagg taattaaata 2761 agcttcaaat aaaacgaaag gctcagtcga aagactgggc ctttcgtttt atctgttgtt 2821 tgtcggtgaa cgctctcctg agtaggacaa atccgccggg agcggatttg aacgttgcga 2881 agcaacggcc cggagggtgg cgggcaggac gcccgccata aactgccagg catcaaatta 2941 agcagaaggc catcctgacg gatggccttt ttgcgtttct acaaactctt tcggtccgtt 3001 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa 3061 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta 3121 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag 3181 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca 3241 gcggtaagat ccttgagagt tttcgccccg aagaacgttt cccaatgatg agcactttta 3301 aagttctgct atgtggcgcg gtattatccc gtgttgacgc cgggcaagag caactcggtc 3361 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc 3421 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca 3481 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc 3541 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca 3601 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac 3661 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg 3721 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg 3781 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg 3841 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac 3901 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc 3961 aagtttactc atatatactt tagattgatt tccttaggac tgagcgtcaa ccccgtagaa 4021 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca 4081 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt 4141 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg 4201 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc 4261 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga 4321 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc 4381 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc 4441 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca 4501 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg 4561 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta 4621 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct 4681 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag 4741 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa 4801 gcggaagagc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc 4861 atataaggtg cactgtgact gggtcatggc tgcgccccga cacccgccaa cacccgctga 4921 cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc 4981 cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga ggcagctgcg 5041 gtaaagctca tcagcgtggt cgtgcagcga ttcacagatg tctgcctgtt catccgcgtc 5101 cagctcgttg agtttctcca gaagcgttaa tgtctggctt ctgataaagc gggccatgtt 5161 aagggcggtt ttttcctgtt tggtcactga tgcctccgtg taagggggat ttctgttcat 5221 gggggtaatg ataccgatga aacgagagag gatgctcacg atacgggtta ctgatgatga 5281 acatgcccgg ttactggaac gttgtgaggg taaacaactg gcggtatgga tgcggcggga 5341 ccagagaaaa atcactcagg gtcaatgcca gcgcttcgtt aatacagatg taggtgttcc 5401 acagggtagc cagcagcatc ctgcgatgca gatccggaac ataatggtgc agggcgctga 5461 cttccgcgtt tccagacttt acgaaacacg gaaaccgaag accattcatg ttgttgctca 5521 ggtcgcagac gttttgcagc agcagtcgct tcacgttcgc tcgcgtatcg gtgattcatt 5581 ctgctaacca gtaaggcaac cccgccagcc tagccgggtc ctcaacgaca ggagcacgat 5641 catgcgcacc cgtggccagg acccaacgct gcccgaaatt //