LOCUS       sequence-146382-        8123 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 8123)
  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 8123)
  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: pNXRVbt-HACre
FEATURES             Location/Qualifiers
     source          1..8123
                     /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"
     terminator      5157..5354
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     rep_origin      5409..6192
                     /label=ars1
                     /note="Schizosaccharomyces pombe autonomously replicating 
                     sequence ars1"
     primer_bind     complement(6203..6220)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6374..6962)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6454..6473)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7133..7924,7925..7993))
                     /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     7756..7775
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7994..8098)
                     /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 tcttctagaa agatagttat actagtcata
     2341 tgcctacaac aactaatggc tatcatgcgg aagctggtga gaagaacagc atcgggacaa
     2401 gggaaggaag aacaaagaca aagaaaacaa aagaaagcaa ttgaaaacaa aacaaaacaa
     2461 ttttcattcc ttctcttatc attccttttc ttttcttttc tctcattcaa cgcactccat
     2521 cgtatccgta ttcctcttat tttttctctt tctctatatc catttctttc tctctaggtg
     2581 tgtcctctct ctctcttcaa tttctctact ccgcattcca acgcatcctt cccccaacct
     2641 cccatttcct ccttacggcc cgatagcgat cgtctttccc tcgctatcac tcgctaccgg
     2701 cccctcctct gcaccgtaac ctcctacgta tttaccatat cataaagttt tttccgacgc
     2761 ttatcgctga ccccctgtcg ccctcctatt ggcttccgga ttatcttctt gtccataagg
     2821 tgatccatgc ttcctgaaga ttcccgaaat gtgtccactt tggcggggaa tcattccatc
     2881 cacttctttc tctctcgctt tcctcattcg gcgctcccct tccgcgtctc attggtcttc
     2941 cgctccgttt ttgctttgcc gatgttactt ggggagaggt gcgataatcc tttcgcaaaa
     3001 actcggtttg acgcctccca tggtataaat agtgggtggt ggacaggtgc cttcgctttt
     3061 cttttcttta agcaagagaa gcttatgaaa aagcctgaac tcaccgcgac gtctgtcgag
     3121 aagtttctga tcgaaaagtt cgacagcgtc tccgacctga tgcagctctc ggagggcgaa
     3181 gaatctcgtg ctttcagctt cgatgtagga gggcgtggat atgtcctgcg ggtaaatagc
     3241 tgcgccgatg gtttctacaa agatcgttat gtttatcggc actttgcatc ggccgcgctc
     3301 ccgattccgg aagtgcttga cattggggaa ttcagcgaga gcctgaccta ttgcatctcc
     3361 cgccgtgcac agggtgtcac gttgcaagac ctgcctgaaa ccgaactgcc cgctgttctg
     3421 cagccggtcg cggaggccat ggatgcgatc gctgcggccg atcttagcca gacgagcggg
     3481 ttcggcccat tcggaccgca aggaatcggt caatacacta catggcgtga tttcatatgc
     3541 gcgattgctg atccccatgt gtatcactgg caaactgtga tggacgacac cgtcagtgcg
     3601 tccgtcgcgc aggctctcga tgagctgatg ctttgggccg aggactgccc cgaagtccgg
     3661 cacctcgtgc acgcggattt cggctccaac aatgtcctga cggacaatgg ccgcataaca
     3721 gcggtcattg actggagcga ggcgatgttc ggggattccc aatacgaggt cgccaacatc
     3781 ttcttctgga ggccgtggtt ggcttgtatg gagcagcaga cgcgctactt cgagcggagg
     3841 catccggagc ttgcaggatc gccgcggctc cgggcgtata tgctccgcat tggtcttgac
     3901 caactctatc agagcttggt tgacggcaat ttcgatgatg cagcttgggc gcagggtcga
     3961 tgcgacgcaa tcgtccgatc cggagccggg actgtcgggc gtacacaaat cgcccgcaga
     4021 agcgcggccg tctggaccga tggctgtgta gaagtagcta gtgcgttcga ccaggctgcg
     4081 cgttctcgcg gccatagcaa ccgacgtacg gcgttgcgcc ctcgccggca gcaagaagcc
     4141 acggaagtcc gcctggagca gaaaatgccc acgctactgc gggtttatat agacggtcct
     4201 cacgggatgg ggaaaaccac caccacgcaa ctgctggtgg ccctgggttc gcgcgacgat
     4261 atcgtctacg tacccgagcc gatgacttac tggcaggtgc tgggggcttc cgagacaatc
     4321 gcgaacatct acaccacaca acaccgcctc gaccagggtg agatatcggc cggggacgcg
     4381 gcggtggtaa tgacaagcgc ccagataaca atgggcatgc cttatgccgt gaccgacgcc
     4441 gttctggctc ctcatatcgg gggggaggct gggagctcac atgccccgcc cccggccctc
     4501 accctcatct tcgaccgcca tcccatcgcc gccctcctgt gctacccggc cgcgcgatac
     4561 cttatgggca gcatgacccc ccaggccgtg ctggcgttcg tggccctcat cccgccgacc
     4621 ttgcccggca caaacatcgt gttgggggcc cttccggagg acagacacat cgaccgcctg
     4681 gccaaacgcc agcgccccgg cgagcggctt gacctggcta tgctggccgc gattcgccgc
     4741 gtttacgggc tgcttgccaa tacggtgcgg tatctgcagg gcggcgggtc gtggtgggag
     4801 gattggggac agctttcggg gacggccgtg ccgccccagg gtgccgagcc ccagagcaac
     4861 gcgggcccac gaccccatat cggggacacg ttatttaccc tgtttcgggc ccccgagttg
     4921 ctggccccca acggcgacct gtataacgtg tttgcctggg ccttggacgt cttggccaaa
     4981 cgcctccgtc ccatgcacgt ctttatcctg gattacgacc aatcgcccgc cggctgccgg
     5041 gacgccctgc tgcaacttac ctccgggatg gtccagaccc acgtcaccac cccaggctcc
     5101 ataccgacga tctgcgacct ggcgcgcacg tttgcccggc cctgccccta atcagtactg
     5161 acaataaaaa gattcttgtt ttcaagaact tgtcatttgt atagtttttt tatattgtag
     5221 ttgttctatt ttaatcaaat gttagcgtga tttatatttt ttttcgcctc gacatcatct
     5281 gcccagatgc gaagttaagt gcgcagaaag taatatcatg cgtcaatcgt atgtgaatgc
     5341 tggtcgctat actgctgtcg attcgatact aacgccgcca tccagtttaa acgagctcga
     5401 attctaggta ggcattttgt ttagttaaag gtacttaagt aattagtata aacgaaccaa
     5461 ttttataatc aggaagttaa gtgaatggta gcacatgtcg taaaaattgt gaatttttat
     5521 tgaataatat tttaaataca agcctttcta gactaggtat actcataaac atatatgagc
     5581 aaaaggatag aggagattac attgcatctt ctacaaattt atttattgcc ctttactgaa
     5641 aaattaaata atgagtacta aatgataaaa agcgctcagt acaagaaaga ttgcaaaaat
     5701 attgcattct tcatgaatta aagttgcata taaggcatat tgaaagtaat agtactaaaa
     5761 cagcagttag cgaaaattaa tagaattata ttcgcaagac aattgtgaca ttaaattaaa
     5821 aaattgtaga atttttacta tcctctttaa cgccatgagc ctttataaaa aggttaaatt
     5881 agttttaaca ttcttttttt gagtaagagt ttacaattta tcaaaacctg tgttattata
     5941 ttcattagtt tcaatttatt agcatctaga gaaaaatcaa ttggcagtta cattgttgga
     6001 atttatgaag aaaaagaatc tacaacggag aataagttgc tgatcgcttt ccctaaaatt
     6061 gtatattttg ctgagcttat tttgacattt cgttgaagtt ttctctaatt cgcattcatt
     6121 ttaagtaaaa caatgagaaa taaaattaca aaaaatacaa attaaaatac aatttttagc
     6181 tataattata gacctaggct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     6241 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     6301 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     6361 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     6421 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     6481 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     6541 tcccttcggg aagcgtggcg ctttctcaat gctcacgctg taggtatctc agttcggtgt
     6601 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     6661 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     6721 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     6781 tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc
     6841 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     6901 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     6961 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     7021 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     7081 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     7141 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     7201 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     7261 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     7321 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     7381 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     7441 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     7501 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     7561 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     7621 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     7681 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     7741 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     7801 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     7861 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     7921 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     7981 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     8041 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     8101 catttccccg aaaagtgcca cct
//
