LOCUS pMAL-cRI 6130 bp DNA circular 06-JUN-2014 DEFINITION Cloning vector pMAL-cRI, complete sequence. ACCESSION VERSION KEYWORDS . SOURCE Cloning vector pMAL-cRI ORGANISM Cloning vector pMAL-cRI other sequences; artificial sequences; vectors. REFERENCE 1 (bases 1 to 6130) 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 6130) 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..6130 /organism="Cloning vector pMAL-cRI" /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..3292 /gene="malE expression cassette" promoter 1406..1433 /gene="malE expression cassette" /note="Ptac promoter (clockwise)" CDS 1528..2898 /gene="malE expression cassette" /note="malE sequence 1528..2628; multiple cloning site region 2629..2715; lacZalpha sequence 2716..2897" /codon_start=1 /product="maltose binding protein (MBP) fusion" /translation="MKTEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKL EEKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNG KLIAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWP LIAADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAF NKGETAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKEL AKEFLENYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNI PQMSAFWYAVRTAVINAASGRQTVDEALKDAQTNSSSVPSSLVIEGRISEFGSSRVDL QASLALAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWQ LGCFGG" primer_bind 2587..2610 /note="malE primer sequence" misc_feature 2634..2714 /gene="malE expression cassette" /note="multiple cloning site (SacI-HindIII)" terminator 3086..3288 /gene="malE expression cassette" /note="rrnB T1T2 transcription terminators" gene 3399..4259 /gene="bla" CDS 3399..4259 /gene="bla" /note="ampR (confers resistance to ampicillin)" /codon_start=1 /product="beta-lactamase" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVE YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA EIGASLIKHW" sig_peptide 3399..3467 /gene="bla" /note="required for secretion to the periplasm; cleaved off to form the mature beta-lactamase protein" rep_origin 4430..5018 /note="pMB1 origin of replication (clockwise) (RNAII -35 to RNA/DNA switch point)" gene complement(5446..5637) /gene="rop" CDS complement(5446..5637) /gene="rop" /exception="alternative start codon" /codon_start=1 /transl_table=11 /product="ROP protein" /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDH ADELYRSCLARFGDDGENL" BASE COUNT 1518 a 1556 c 1639 g 1417 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 aaaactgaag 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 agctggcgaa agagttcctc gaaaactatc tgctgactga tgaaggtctg 2401 gaagcggtta ataaagacaa accgctgggt gccgtagcgc tgaagtctta cgaggaagag 2461 ttggcgaaag atccacgtat tgccgccacc atggaaaacg cccagaaagg tgaaatcatg 2521 ccgaacatcc cgcagatgtc cgctttctgg tatgccgtgc gtactgcggt gatcaacgcc 2581 gccagcggtc gtcagactgt cgatgaagcc ctgaaagacg cgcagactaa ttcgagctcg 2641 gtaccgtcct ctctcgtgat cgagggaagg atttcagaat tcggatcctc tagagtcgac 2701 ctgcaggcaa gcttggcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc 2761 gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa 2821 gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atggcagctt 2881 ggctgttttg gcggatgaga gaagattttc agcctgatac agattaaatc agaacgcaga 2941 agcggtctga taaaacagaa tttgcctggc ggcagtagcg cggtggtccc acctgacccc 3001 atgccgaact cagaagtgaa acgccgtagc gccgatggta gtgtggggtc tccccatgcg 3061 agagtaggga actgccaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt 3121 tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc cgccgggagc 3181 ggatttgaac gttgcgaagc aacggcccgg agggtggcgg gcaggacgcc cgccataaac 3241 tgccaggcat caaattaagc agaaggccat cctgacggat ggcctttttg cgtttctaca 3301 aactcttttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac 3361 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg 3421 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc 3481 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg 3541 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga 3601 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc 3661 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag 3721 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga 3781 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg 3841 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga 3901 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt 3961 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact 4021 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt 4081 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg 4141 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta 4201 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac 4261 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta 4321 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt 4381 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt 4441 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt 4501 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc 4561 agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg 4621 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg 4681 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt 4741 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac 4801 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg 4861 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg 4921 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat 4981 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt 5041 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg 5101 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa 5161 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc 5221 tccttacgca tctgtgcggt atttcacacc gcatatggtg cactctcagt acaatctgct 5281 ctgatgccgc atagttaagc cagtatacac tccgctatcg ctacgtgact gggtcatggc 5341 tgcgccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc 5401 atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc 5461 gtcatcaccg aaacgcgcga ggcagctgcg gtaaagctca tcagcgtggt cgtgaagcga 5521 ttcacagatg tctgcctgtt catccgcgtc cagctcgttg agtttctcca gaagcgttaa 5581 tgtctggctt ctgataaagc gggccatgtt aagggcggtt ttttcctgtt tggtcactga 5641 tgcctccgtg taagggggat ttctgttcat gggggtaatg ataccgatga aacgagagag 5701 gatgctcacg atacgggtta ctgatgatga acatgcccgg ttactggaac gttgtgaggg 5761 taaacaactg gcggtatgga tgcggcggga ccagagaaaa atcactcagg gtcaatgcca 5821 gcgcttcgtt aatacagatg taggtgttcc acagggtagc cagcagcatc ctgcgatgca 5881 gatccggaac ataatggtgc agggcgctga cttccgcgtt tccagacttt acgaaacacg 5941 gaaaccgaag accattcatg ttgttgctca ggtcgcagac gttttgcagc agcagtcgct 6001 tcacgttcgc tcgcgtatcg gtgattcatt ctgctaacca gtaaggcaac cccgccagcc 6061 tagccgggtc ctcaacgaca ggagcacgat catgcgcacc cgtggccagg acccaacgct 6121 gcccgaaatt //