LOCUS       sequence-146386-        6493 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  constitutive Cre expression vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
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 Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "BMC 
            Biotechnol."; date: "2016-11-8"; volume: "16(1)"; pages: "76"
COMMENT     Sequence Label: pNXRVh-HACre
FEATURES             Location/Qualifiers
     source          1..6493
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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(join(5503..6294,6295..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
//
