LOCUS pMAL-pIII 6706 bp DNA circular 12-MAR-2014 DEFINITION Cloning vector pMAL-pIII, complete sequence. ACCESSION VERSION KEYWORDS . SOURCE Cloning vector pMAL-pIII ORGANISM Cloning vector pMAL-pIII other sequences; artificial sequences; vectors. REFERENCE 1 (bases 1 to 6706) AUTHORS Zagursky,R.J. and Berman,M.L. TITLE Cloning vectors that yield high levels of single-stranded DNA for rapid DNA sequencing JOURNAL Gene 27 (2), 183-191 (1984) PUBMED 6327466 REFERENCE 2 (bases 1 to 6706) AUTHORS Zwick,M.B., Bonnycastle,L.L., Noren,K.A., Venturini,S., Leong,E., Barbas,C.F. III, Noren,C.J. and Scott,J.K. TITLE The maltose-binding protein as a scaffold for monovalent display of peptides derived from phage libraries JOURNAL Anal Biochem 264 (1), 87-97 (1998) PUBMED 9784192 REFERENCE 3 (bases 1 to 6706) AUTHORS Paschal,B.M. TITLE Direct Submission JOURNAL Submitted (22-OCT-2007) Research Department, New England Biolabs, 240 County Road, Ipswich, MA 01938, USA FEATURES Location/Qualifiers source 1..6706 /organism="Cloning vector pMAL-pIII" /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..2976 /gene="malE expression cassette" /note="pIII leader sequence 1528..1581; malE sequence 1591..2688; multiple cloning site region 2689..2794; lacZalpha sequence 2795..2976" /codon_start=1 /product="pIII/maltose binding protein (MBP) fusion" /translation="MKKLLFAIPLVVPFYSHSADMKTEEGKLVIWINGDKGYNGLAEV GKKFEKDTGIKVTVEHPDKLEEKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITP DKAFQDKLYPFTWDAVRYNGKLIAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKEL KAKGKSALMFNLQEPYFTWPLIAADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVD LIKNKHMNADTDYSIAEAAFNKGETAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPS KPFVGVLSAGINAASPNKELAKEFLENYLLTDEGLEAVNKDKPLGAVALKSYEEELAK DPRIAATMENAQKGEIMPNIPQMSAFWYAVRTAVINAASGRQTVDEALKDAQTNSSSN NNNNNNNNNLGIEGRISEFGSSRVDLQASLALAVVLQRRDWENPGVTQLNRLAAHPPF ASWRNSEEARTDRPSQQLRSLNGEWQLGCFGG" sig_peptide 1528..1581 /gene="malE expression cassette" /note="pIII leader sequence (required for secretion to the periplasm; cleaved off to form the mature MBP or MPB-target peptide fusion)" primer_bind 2647..2670 /note="malE primer sequence" misc_feature 2770..2807 /gene="malE expression cassette" /note="multiple cloning site (EcoRI-HindIII); not indended for use" terminator 3164..3367 /note="rrnB T1T2 transcription terminators" gene 3478..4338 /gene="bla" CDS 3478..4338 /gene="bla" /note="ampR (confers resistance to ampicillin)" /codon_start=1 /product="beta-lactamase" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVE YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA EIGASLIKHW" sig_peptide 3478..3546 /gene="bla" /note="required for secretion to the periplasm; cleaved off to form the mature beta-lactamase protein" rep_origin complement(4380..4893) /note="M13 origin of replication (+ -)" rep_origin 5004..5592 /note="pMB1 origin of replication (clockwise) (RNAII -35 to RNA/DNA switch point)" gene complement(6022..6213) /gene="rop" CDS complement(6022..6213) /gene="rop" /exception="alternative start codon" /codon_start=1 /transl_table=11 /product="ROP protein" /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDH ADELYRSCLARFGDDGENL" BASE COUNT 1711 a 1683 c 1775 g 1537 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 aaaaaattat tattcgcaat tcctttagtg 1561 gtacctttct attctcactc ggccgatatg aaaactgaag aaggtaaact ggtaatctgg 1621 attaacggcg ataaaggcta taacggtctc gctgaagtcg gtaagaaatt cgagaaagat 1681 accggaatta aagtcaccgt tgagcatccg gataaactgg aagagaaatt cccacaggtt 1741 gcggcaactg gcgatggccc tgacattatc ttctgggcac acgaccgctt tggtggctac 1801 gctcaatctg gcctgttggc tgaaatcacc ccggacaaag cgttccagga caagctgtat 1861 ccgtttacct gggatgccgt acgttacaac ggcaagctga ttgcttaccc gatcgctgtt 1921 gaagcgttat cgctgattta taacaaagat ctgctgccga acccgccaaa aacctgggaa 1981 gagatcccgg cgctggataa agaactgaaa gcgaaaggta agagcgcgct gatgttcaac 2041 ctgcaagaac cgtacttcac ctggccgctg attgctgctg acgggggtta tgcgttcaag 2101 tatgaaaacg gcaagtacga cattaaagac gtgggcgtgg ataacgctgg cgcgaaagcg 2161 ggtctgacct tcctggttga cctgattaaa aacaaacaca tgaatgcaga caccgattac 2221 tccatcgcag aagctgcctt taataaaggc gaaacagcga tgaccatcaa cggcccgtgg 2281 gcatggtcca acatcgacac cagcaaagtg aattatggtg taacggtact gccgaccttc 2341 aagggtcaac catccaaacc gttcgttggc gtgctgagcg caggtattaa cgccgccagt 2401 ccgaacaaag agctggcaaa agagttcctc gaaaactatc tgctgactga tgaaggtctg 2461 gaagcggtta ataaagacaa accgctgggt gccgtagcgc tgaagtctta cgaggaagag 2521 ttggcgaaag atccacgtat tgccgccact atggaaaacg cccagaaagg tgaaatcatg 2581 ccgaacatcc cgcagatgtc cgctttctgg tatgccgtgc gtactgcggt gatcaacgcc 2641 gccagcggtc gtcagactgt cgatgaagcc ctgaaagacg cgcagactaa ttcgagctcg 2701 aacaacaaca acaataacaa taacaacaac ctcgggatcg agggaaggat ttcagaattc 2761 ggatcctcta gagtcgacct gcaggcaagc ttggcactgg ccgtcgtttt acaacgtcgt 2821 gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc 2881 agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg 2941 aatggcgaat ggcagcttgg ctgttttggc ggatgagata agattttcag cctgatacag 3001 attaaatcag aacgcagaag cggtctgata aaacagaatt tgcctggcgg cagtagcgcg 3061 gtggtcccac ctgaccccat gccgaactca gaagtgaaac gccgtagcgc cgatggtagt 3121 gtggggtctc cccatgcgag agtagggaac tgccaggcat caaataaaac gaaaggctca 3181 gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc tcctgagtag 3241 gacaaatccg ccgggagcgg atttgaacgt tgcgaagcaa cggcccggag ggtggcgggc 3301 aggacgcccg ccataaactg ccaggcatca aattaagcag aaggccatcc tgacggatgg 3361 cctttttgcg tttctacaaa ctctttttgt ttatttttct aaatacattc aaatatgtat 3421 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg 3481 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt 3541 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga 3601 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa 3661 gaacgttctc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt 3721 gttgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt 3781 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc 3841 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga 3901 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat 3961 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct 4021 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc 4081 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg 4141 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc 4201 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg 4261 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca 4321 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta 4381 ccccggttga taatcagaaa agccccaaaa acaggaagat tgtataagca aatatttaaa 4441 ttgtaaacgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt 4501 ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag 4561 ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg 4621 tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcacccaaat 4681 caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc 4741 gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga 4801 aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac 4861 ccgccgcgct taatgcgccg ctacagggcg cgtaaaagga tctaggtgaa gatccttttt 4921 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc 4981 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg 5041 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact 5101 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg 5161 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg 5221 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac 5281 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca 5341 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga 5401 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc 5461 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct 5521 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg 5581 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct 5641 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc 5701 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc 5761 gaggaagcgg aagagcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca 5821 caccgcatat atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagt 5881 atacactccg ctatcgctac gtgactgggt catggctgcg ccccgacacc cgccaacacc 5941 cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac 6001 cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca 6061 gctgcggtaa agctcatcag cgtggtcgtg cagcgattca cagatgtctg cctgttcatc 6121 cgcgtccagc tcgttgagtt tctccagaag cgttaatgtc tggcttctga taaagcgggc 6181 catgttaagg gcggtttttt cctgtttggt cactgatgcc tccgtgtaag ggggatttct 6241 gttcatgggg gtaatgatac cgatgaaacg agagaggatg ctcacgatac gggttactga 6301 tgatgaacat gcccggttac tggaacgttg tgagggtaaa caactggcgg tatggatgcg 6361 gcgggaccag agaaaaatca ctcagggtca atgccagcgc ttcgttaata cagatgtagg 6421 tgttccacag ggtagccagc agcatcctgc gatgcagatc cggaacataa tggtgcaggg 6481 cgctgacttc cgcgtttcca gactttacga aacacggaaa ccgaagacca ttcatgttgt 6541 tgctcaggtc gcagacgttt tgcagcagca gtcgcttcac gttcgctcgc gtatcggtga 6601 ttcattctgc taaccagtaa ggcaaccccg ccagcctagc cgggtcctca acgacaggag 6661 cacgatcatg cgcacccgtg gccaggaccc aacgctgccc gaaatt //