LOCUS Exported 2817 bp ds-DNA circular SYN 25-11-2013 DEFINITION High copy-number E. coli vector with a multiple cloning site flanked by mutant T7 promoters. The MCS is reversed relative to LITMUS38. ACCESSION . VERSION . KEYWORDS LITMUS39 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 2817) AUTHORS Evans PD, Cook SN, Riggs PD, Noren CJ. TITLE LITMUS: multipurpose cloning vectors with a novel system for bidirectional in vitro transcription. JOURNAL BioTechniques 1995;19:130-5. PUBMED 7669286 REFERENCE 2 (bases 1 to 2817) AUTHORS New England Biolabs TITLE Direct Submission FEATURES Location/Qualifiers source 1..2817 /organism="synthetic DNA construct" /lab_host="Escherichia coli" /mol_type="other DNA" promoter 38..142 /gene="bla" /note="AmpR promoter" CDS 143..1003 /codon_start=1 /gene="bla" /product="beta-lactamase" /note="AmpR" /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERSPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" rep_origin complement(1045..1554) /direction=LEFT /note="M13 ori" /note="M13 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" rep_origin 1665..2253 /direction=RIGHT /note="ori" /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" promoter 2350..2380 /note="lac promoter" /note="promoter for the E. coli lac operon" protein_bind 2388..2404 /bound_moiety="lac repressor encoded by lacI" /note="lac operator" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." primer_bind 2412..2428 /note="M13 rev" /note="common sequencing primer, one of multiple similar variants" CDS 2424..2789 /codon_start=1 /gene="lacZalpha" /product="LacZ-alpha fragment of beta-galactosidase" /note="lacZ-alpha" /translation="MTMITPSYVIRLTSGSTACLYTPEDAFGRASEFWIHDILQEAFWR LAGAIEWAPLVKALSESYYGLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEARTD RPSQQLRSLNGEWRFAW" promoter 2449..2466 /gene="lacZalpha" /note="T7 promoter (mutant)" /note="mutant version of the promoter for bacteriophage T7 RNA polymerase" misc_feature 2460..2602 /gene="lacZalpha" /note="MCS" /note="multiple cloning site" promoter complement(2597..2614) /note="T7 promoter (mutant)" /note="mutant version of the promoter for bacteriophage T7 RNA polymerase" primer_bind complement(2618..2634) /note="M13 fwd" /note="common sequencing primer, one of multiple similar variants" ORIGIN 1 gttaactacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt 61 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca 121 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt 181 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga 241 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa 301 gatccttgag agttttcgcc ccgaagaacg ttctccaatg atgagcactt ttaaagttct 361 gctatgtggc gcggtattat cccgtgttga cgccgggcaa gagcaactcg gtcgccgcat 421 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga 481 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc 541 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat 601 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa 661 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac 721 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa 781 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc 841 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc 901 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag 961 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta 1021 ctcatatata ctttagattg atttaccccg gttgataatc agaaaagccc caaaaacagg 1081 aagattgtat aagcaaatat ttaaattgta aacgttaata ttttgttaaa attcgcgtta 1141 aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat 1201 aaatcaaaag aatagcccga gatagggttg agtgttgttc cagtttggaa caagagtcca 1261 ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc 1321 ccactacgtg aaccatcacc caaatcaagt tttttggggt cgaggtgccg taaagcacta 1381 aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcg aacgtggcga 1441 gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca 1501 cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtaaaagg 1561 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg 1621 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt 1681 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg 1741 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata 1801 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca 1861 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag 1921 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc 1981 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga 2041 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg 2101 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac 2161 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg 2221 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg 2281 ttcctggcct tttgctggcc ttttgctcac atgtaatgtg agttagctca ctcattaggc 2341 accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata 2401 acaatttcac acaggaaaca gctatgacca tgattacgcc aagctacgta atacgactca 2461 ctagtgggtc gaccgcatgc ctgtacactc cggaagacgc gttcggccgt gctagcgaat 2521 tctggatcca cgatatcctg caggaagctt tctggcgcct ggccggcgca attgaatggg 2581 cccctctagt caaggcctta agtgagtcgt attacggact ggccgtcgtt ttacaacgtc 2641 gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg 2701 ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc 2761 tgaatggcga atggcgcttc gcttggtaat aaagcccgct tcggcgggct ttttttt //