LOCUS pMiniT2.0 2588 bp DNA circular 07-SEP-2016 DEFINITION Cloning vector pMiniT 2.0, complete sequence. ACCESSION VERSION KEYWORDS . SOURCE Cloning vector pMiniT2.0 ORGANISM Cloning vector pMiniT2.0 other sequences; artificial sequences; vectors. REFERENCE 1 (bases 1 to 2588) AUTHORS Kucera,R.B. TITLE Direct Submission JOURNAL Submitted (07-SEP-2016) Research Department, New England Biolabs, 240 County Road, Ipswich, MA 01938, USA FEATURES Location/Qualifiers source 1..2588 /organism="Cloning vector pMiniT2.0" /mol_type="other DNA" promoter 1..214 /note="constitutive promoter region" primer_bind 391..412 /note="cloning analysis forward primer sequence" promoter 479..496 /note="SP6 promoter (transcript forward)" gene 541..549 /gene="toxic minigene" CDS 541..549 /gene="toxic minigene" /note="2 additional stop codons beyond gene boundary" /codon_start=1 /product="Met-Ile" /translation="MI" misc_feature 545..546 /gene="toxic minigene" /note="blunt end cleavage site; product is supplied in linearized form, cut between these two bases" promoter complement(602..619) /note="v2 T7 promoter (transcript start 602 reverse)" primer_bind complement(677..699) /note="cloning analysis reverse primer sequence" gene 733..1593 /gene="bla" CDS 733..1593 /gene="bla" /note="ampR (confers resistance to ampicillin) (mutant V82I,A182V)" /codon_start=1 /product="beta-lactamase" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA EIGASLIKHW" rep_origin 1764..2352 /note="pUC19 origin of replication (RNAII -35 to RNA/DNA switch point)" BASE COUNT 702 a 616 c 617 g 653 t ORIGIN 1 ccggttcaga caggataaag aggaacgcag aatgttagac aacacccgct tacgcatagc 61 tattcagaaa tcaggccgtt taagcgatga ttcacgagaa ttgctggccc gctgcggcat 121 aaaaattaat ttacacactc agcgctgatg aatcccctaa tgattttggt aaaaatcatt 181 aagttaaggt ggacacacat cttgtcatat gattaaatgg tttcgcgaaa aatcaataat 241 cagacaacaa gatgtgcgaa ctcgatattt tacacgactc tctttaccaa ttctgccccg 301 aattacactt aaaacgactc aacagcttaa cgttggcttg ccacgcatta cttgactgta 361 aaactctcac tcttaccgaa cttggccgta acctgccaac caaagcgaga acaaaacata 421 acatcaaacg aatcgaccga ttgttaggta atcgtcacct gcaggaaggt ttaaacgcat 481 ttaggtgaca ctatagaagt gtgtatcgct cgagggatcc gaattcagga ggtaaaaacc 541 atgatctgat aataattaat taagacgtca gaattctcga ggcggccgca tgtgcgtctc 601 cctatagtga gtcgtattaa tttcgcgggc ggaaccccta tttgtttatt tttctaaata 661 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga 721 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca 781 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat 841 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag 901 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc 961 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct 1021 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca 1081 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt 1141 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat 1201 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt 1261 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta 1321 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga 1381 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt 1441 gagcgtggat ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc 1501 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct 1561 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata 1621 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt 1681 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc 1741 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg 1801 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact 1861 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg 1921 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg 1981 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac 2041 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca 2101 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga 2161 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc 2221 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct 2281 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg 2341 agcctatgga aaaacgccag caatgcggcc tttttacggt tcctggcctt ttgctggcct 2401 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc 2461 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc 2521 gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat 2581 taatgcag //