LOCUS Exported 6493 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION constitutive Cre expression vector. ACCESSION . VERSION . KEYWORDS pNXRVh-HACre SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6493) AUTHORS Amelina H, Moiseeva V, Collopy LC, Pearson SR, Armstrong CA, Tomita K TITLE Sequential and counter-selectable cassettes for fission yeast. JOURNAL BMC Biotechnol. 2016 Nov 8;16(1):76. PUBMED 27825338 REFERENCE 2 (bases 1 to 6493) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6493 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 43..61 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" primer_bind complement(99..121) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 221..240 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 295..598 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 599..802 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 753..773 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" CDS 895..921 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" CDS 925..951 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" CDS 958..984 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" CDS 1003..1023 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 1033..2061 /codon_start=1 /product="site-specific recombinase" /label=Cre /note="Cre is a site-specific recombinase from bacteriophage P1. Recombination occurs at loxP sequences." /translation="MANLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKML LSVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGL PRPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAF LGIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVE RWISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQ RYLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLE DGD" terminator 2091..2278 /gene="S. cerevisiae ADH1" /label=ADH1 terminator /note="transcription terminator for the S. cerevisiae alcohol dehydrogenase 1 (ADH1) gene" primer_bind complement(2146..2170) /label=pHybLex-R /note="ADH terminator, reverse primer" rep_origin 3779..4562 /label=ars1 /note="Schizosaccharomyces pombe autonomously replicating sequence ars1" primer_bind complement(4573..4590) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4744..5332) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4824..4843) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5503..6363) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 6126..6145 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6364..6468) /gene="bla" /label=AmpR promoter ORIGIN 1 gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg 61 ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg 121 gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg 181 tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta 241 ctgagagtgc accattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac 301 ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc 361 aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt 421 ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac 481 gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac 541 cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt 601 gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc 661 aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt 721 tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg 781 ggaggtctat ataagcagag ctggtttagt gaaccgtcag atccgctagg atctcgagct 841 caagcttcgt acgctgcagg tcgacggatc cccgggttaa ttaacgctag catgtaccca 901 tacgatgttc ctgactatgc gggctacccg tatgacgtgc cggactatgc aggagcctat 961 ccttatgacg ttccagatta cgctggttct agatgtactc caccaaagaa gaagagaaag 1021 gttgccccta gcatggccaa tttactgacc gtacaccaaa atttgcctgc attaccggtc 1081 gatgcaacga gtgatgaggt tcgcaagaac ctgatggaca tgttcaggga tcgccaggcg 1141 ttttctgagc atacctggaa aatgcttctg tccgtttgcc ggtcgtgggc ggcatggtgc 1201 aagttgaata accggaaatg gtttcccgca gaacctgaag atgttcgcga ttatcttcta 1261 tatcttcagg cgcgcggtct ggcagtaaaa actatccagc aacatttggg ccagctaaac 1321 atgcttcatc gtcggtccgg gctgccacga ccaagtgaca gcaatgctgt ttcactggtt 1381 atgcggcgga tccgaaaaga aaacgttgat gccggtgaac gtgcaaaaca ggctctagcg 1441 ttcgaacgca ctgatttcga ccaggttcgt tcactcatgg aaaatagcga tcgctgccag 1501 gatatacgta atctggcatt tctggggatt gcttataaca ccctgttacg tatagccgaa 1561 attgccagga tcagggttaa agatatctca cgtactgacg gtgggagaat gttaatccat 1621 attggcagaa cgaaaacgct ggttagcacc gcaggtgtag agaaggcact tagcctgggg 1681 gtaactaaac tggtcgagcg atggatttcc gtctctggtg tagctgatga tccgaataac 1741 tacctgtttt gccgggtcag aaaaaatggt gttgccgcgc catctgccac cagccagcta 1801 tcaactcgcg ccctggaagg gatttttgaa gcaactcatc gattgattta cggcgctaag 1861 gatgactctg gtcagagata cctggcctgg tctggacaca gtgcccgtgt cggagccgcg 1921 cgagatatgg cccgcgctgg agtttcaata ccggagatca tgcaagctgg tggctggacc 1981 aatgtaaata ttgtcatgaa ctatatccgt aacctggata gtgaaacagg ggcaatggtg 2041 cgcctgctgg aagatggcga ttctagatga ggcgcgccac ttctaaataa gcgaatttct 2101 tatgatttat gatttttatt attaaataag ttataaaaaa aataagtgta tacaaatttt 2161 aaagtgactc ttaggtttta aaacgaaaat tcttattctt gagtaactct ttcctgtagg 2221 tcaggttgct ttctcaggta tagtatgagg tcgctcttat tgaccacacc tctaccggca 2281 gatccgctag ggataacagg gtaatataga tctccatacg atatcccaat ctgtagtata 2341 aaaagtgcaa cagtgttgag tttcccgaaa ccaggcattt ttgcggtagg gtctgaaact 2401 ttcacgaaac catttcacaa caaaacttct tcaacaacat gtgtgcaaag aaaatcgtcg 2461 tcttaccagg agaccatatt ggccctgaaa ttgttgcttc tgccttggag gttttgaaag 2521 tcgttgagaa gaagcgacct gagttaaaac tcgagtttga agaacacaag attggaggtg 2581 cctctattga tgcctatgga acccctttga ctgacgagac tgtgaaggct tgtttggaag 2641 ctgacggtgt tcttttgggt gccgttggtg gtcctgaatg gaccaacccc aattgtcgtc 2701 ctgagcaagg tttattgaag cttcgtaaga gtatgggtgt ttgggccaac cttcgacctt 2761 gcaactttgc cagcaagtct ttagtcaagt acagcccttt gaagcctgaa atcgttgaag 2821 gtgtcgattt ttgtgttgta cgagaactta ctggaggttg ttactttggt gagcgcactg 2881 aggacaacgg atcgggttat gctatggaca cttggcctta cagtttggaa gaagtttctc 2941 gtattgctcg tttggctgct tggttagctg aaacttccaa ccctcctgct cccgtcacat 3001 tactcgacaa agctaatgtt ttggcaactt ctcgtctttg gcgcaagact gtagctaaga 3061 tttttaagga ggaatatcct caccttacct taaaaaacca gctcattgac tccgcagcca 3121 tgcttttggt caagagccct cgtacactta acggtgttgt tttgactgac aacttgtttg 3181 gtgacattat ttcagatgag gcttctgtca ttcctggtag cttgggcctt ttgccttctg 3241 cctccctttc cggtgtggta ggaaaatcag aagaaaaggt tcattgtttg gttgagccca 3301 ttcacggtag cgctcccgat atcgctggca agggcattgt taatcctgtt ggtacaattt 3361 tatctgcttc ccttctcctt cgttatggtt tgaatgctcc taaggaggct gaagctatcg 3421 aagccgccgt acgcaaggtc ttggatgata cttcaattgg tggacgtggt ctttatactc 3481 gcgatttggg aggtgaggct tctaccgctg atattactaa ggctgttgtt gaagaacttg 3541 aaaaaatttt gtaatagggt ttaatgtaga ataaattcat atgatttgat ttgacctaat 3601 ttttattgat taattgtgtc gcatatttat tatcgttaaa aatgcaattg acaaaagcgt 3661 aaattttaag gctctaatgt tttttctttt ggtatgttat agtcaagcaa agatgtatta 3721 agttgcaaaa gcatggaatt gatttacgac atccgtttaa acgagctcga attctaggta 3781 ggcattttgt ttagttaaag gtacttaagt aattagtata aacgaaccaa ttttataatc 3841 aggaagttaa gtgaatggta gcacatgtcg taaaaattgt gaatttttat tgaataatat 3901 tttaaataca agcctttcta gactaggtat actcataaac atatatgagc aaaaggatag 3961 aggagattac attgcatctt ctacaaattt atttattgcc ctttactgaa aaattaaata 4021 atgagtacta aatgataaaa agcgctcagt acaagaaaga ttgcaaaaat attgcattct 4081 tcatgaatta aagttgcata taaggcatat tgaaagtaat agtactaaaa cagcagttag 4141 cgaaaattaa tagaattata ttcgcaagac aattgtgaca ttaaattaaa aaattgtaga 4201 atttttacta tcctctttaa cgccatgagc ctttataaaa aggttaaatt agttttaaca 4261 ttcttttttt gagtaagagt ttacaattta tcaaaacctg tgttattata ttcattagtt 4321 tcaatttatt agcatctaga gaaaaatcaa ttggcagtta cattgttgga atttatgaag 4381 aaaaagaatc tacaacggag aataagttgc tgatcgcttt ccctaaaatt gtatattttg 4441 ctgagcttat tttgacattt cgttgaagtt ttctctaatt cgcattcatt ttaagtaaaa 4501 caatgagaaa taaaattaca aaaaatacaa attaaaatac aatttttagc tataattata 4561 gacctaggct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt 4621 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa 4681 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc 4741 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag 4801 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt 4861 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg 4921 aagcgtggcg ctttctcaat gctcacgctg taggtatctc agttcggtgt aggtcgttcg 4981 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg 5041 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac 5101 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg 5161 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt 5221 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg 5281 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc 5341 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt 5401 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt 5461 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag 5521 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt 5581 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc 5641 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc 5701 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg 5761 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac 5821 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg 5881 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc 5941 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact 6001 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc 6061 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat 6121 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc 6181 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac 6241 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa 6301 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact 6361 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg 6421 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg 6481 aaaagtgcca cct //