LOCUS       sequence-165380-        6697 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 6697)
  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 6697)
  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: pNXRVb-HACre
FEATURES             Location/Qualifiers
     source          1..6697
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        41..344
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        345..548
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     499..519
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             641..667
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             671..697
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             704..730
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             749..769
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             779..1807
                     /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      1837..2024
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1892..1916)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            2099..3674
                     /label=hphMX6
                     /note="yeast selectable marker conferring hygromycin 
                     resistance"
     promoter        2099..2442
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             2443..3471
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLMHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     terminator      3477..3674
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     rep_origin      3731..4512
                     /label=ars1
                     /note="Schizosaccharomyces pombe autonomously replicating 
                     sequence ars1"
     primer_bind     complement(4523..4540)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4694..5282)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4774..4793)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5453..6244,6245..6313))
                     /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     6076..6095
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6314..6418)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6486..6504
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6542..6564)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6664..6683
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 ttacggggtc attagttcat agcccatata tggagttccg cgttacataa cttacggtaa
       61 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
      121 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
      181 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
      241 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
      301 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc
      361 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca
      421 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta
      481 acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa
      541 gcagagctgg tttagtgaac cgtcagatcc gctaggatct cgagctcaag cttcgtacgc
      601 tgcaggtcga cggatccccg ggttaattaa cgctagcatg tacccatacg atgttcctga
      661 ctatgcgggc tacccgtatg acgtgccgga ctatgcagga gcctatcctt atgacgttcc
      721 agattacgct ggttctagat gtactccacc aaagaagaag agaaaggttg cccctagcat
      781 ggccaattta ctgaccgtac accaaaattt gcctgcatta ccggtcgatg caacgagtga
      841 tgaggttcgc aagaacctga tggacatgtt cagggatcgc caggcgtttt ctgagcatac
      901 ctggaaaatg cttctgtccg tttgccggtc gtgggcggca tggtgcaagt tgaataaccg
      961 gaaatggttt cccgcagaac ctgaagatgt tcgcgattat cttctatatc ttcaggcgcg
     1021 cggtctggca gtaaaaacta tccagcaaca tttgggccag ctaaacatgc ttcatcgtcg
     1081 gtccgggctg ccacgaccaa gtgacagcaa tgctgtttca ctggttatgc ggcggatccg
     1141 aaaagaaaac gttgatgccg gtgaacgtgc aaaacaggct ctagcgttcg aacgcactga
     1201 tttcgaccag gttcgttcac tcatggaaaa tagcgatcgc tgccaggata tacgtaatct
     1261 ggcatttctg gggattgctt ataacaccct gttacgtata gccgaaattg ccaggatcag
     1321 ggttaaagat atctcacgta ctgacggtgg gagaatgtta atccatattg gcagaacgaa
     1381 aacgctggtt agcaccgcag gtgtagagaa ggcacttagc ctgggggtaa ctaaactggt
     1441 cgagcgatgg atttccgtct ctggtgtagc tgatgatccg aataactacc tgttttgccg
     1501 ggtcagaaaa aatggtgttg ccgcgccatc tgccaccagc cagctatcaa ctcgcgccct
     1561 ggaagggatt tttgaagcaa ctcatcgatt gatttacggc gctaaggatg actctggtca
     1621 gagatacctg gcctggtctg gacacagtgc ccgtgtcgga gccgcgcgag atatggcccg
     1681 cgctggagtt tcaataccgg agatcatgca agctggtggc tggaccaatg taaatattgt
     1741 catgaactat atccgtaacc tggatagtga aacaggggca atggtgcgcc tgctggaaga
     1801 tggcgattct agatgaggcg cgccacttct aaataagcga atttcttatg atttatgatt
     1861 tttattatta aataagttat aaaaaaaata agtgtataca aattttaaag tgactcttag
     1921 gttttaaaac gaaaattctt attcttgagt aactctttcc tgtaggtcag gttgctttct
     1981 caggtatagt atgaggtcgc tcttattgac cacacctcta ccggcagatc cgctagggat
     2041 aacagggtaa tatagatctg tttagcttgc ctcgtccccg ccgggtcacc cggtcagcga
     2101 catggaggcc cagaataccc tccttgacag tcttgacgtg cgcagctcag gggcatgatg
     2161 tgactgtcgc ccgtacattt agcccataca tccccatgta taatcatttg catccataca
     2221 ttttgatggc cgcacggcgc gaagcaaaaa ttacggctcc tcgctgcgga cctgcgagca
     2281 gggaaacgct cccctcacag acgcgttgaa ttgtccccac gccgcgcccc tgtagagaaa
     2341 tataaaaggt taggatttgc cactgaggtt cttctttcat atacttcctt ttaaaatctt
     2401 gctaggatac agttctcaca tcacatccga acataaacaa ccatgggtaa aaagcctgaa
     2461 ctcaccgcga cgtctgtcga gaagtttctg atcgaaaagt tcgacagcgt ctccgacctg
     2521 atgcagctct cggagggcga agaatctcgt gctttcagct tcgatgtagg agggcgtgga
     2581 tatgtcctgc gggtaaatag ctgcgccgat ggtttctaca aagatcgtta tgtttatcgg
     2641 cactttgcat cggccgcgct cccgattccg gaagtgcttg acattgggga attcagcgag
     2701 agcctgacct attgcatctc ccgccgtgca cagggtgtca cgttgcaaga cctgcctgaa
     2761 accgaactgc ccgctgttct gcagccggtc gcggaggcca tggatgcgat cgctgcggcc
     2821 gatcttagcc agacgagcgg gttcggccca ttcggaccgc aaggaatcgg tcaatacact
     2881 acatggcgtg atttcatatg cgcgattgct gatccccatg tgtatcactg gcaaactgtg
     2941 atggacgaca ccgtcagtgc gtccgtcgcg caggctctcg atgagctgat gctttgggcc
     3001 gaggactgcc ccgaagtccg gcacctcatg cacgcggatt tcggctccaa caatgtcctg
     3061 acggacaatg gccgcataac agcggtcatt gactggagcg aggcgatgtt cggggattcc
     3121 caatacgagg tcgccaacat cttcttctgg aggccgtggt tggcttgtat ggagcagcag
     3181 acgcgctact tcgagcggag gcatccggag cttgcaggat cgccgcggct ccgggcgtat
     3241 atgctccgca ttggtcttga ccaactctat cagagcttgg ttgacggcaa tttcgatgat
     3301 gcagcttggg cgcagggtcg atgcgacgca atcgtccgat ccggagccgg gactgtcggg
     3361 cgtacacaaa tcgcccgcag aagcgcggcc gtctggaccg atggctgtgt agaagtactc
     3421 gccgatagtg gaaaccgacg ccccagcact cgtccgaggg caaaggaata atcagtactg
     3481 acaataaaaa gattcttgtt ttcaagaact tgtcatttgt atagtttttt tatattgtag
     3541 ttgttctatt ttaatcaaat gttagcgtga tttatatttt ttttcgcctc gacatcatct
     3601 gcccagatgc gaagttaagt gcgcagaaag taatatcatg cgtcaatcgt atgtgaatgc
     3661 tggtcgctat actgctgtcg attcgatact aacgccgcca tccagtgttt aaacgagctc
     3721 gaattctagg taggcatttt gtttagttaa aggtacttaa gtaattagta taaacgaacc
     3781 aattttataa tcaggaagtt aagtgaatgg tagcacatgt cgtaaaaatt gtgaattttt
     3841 attgaataat attttaaata caagcctttc tagactaggt atactcataa acatatgagc
     3901 aaaaggatag aggagattac attgcatctt ctacaaattt atttattgcc ctttactgaa
     3961 aaattaaata atgagtacta aatgataaaa agcgctcagt acaagaaaga ttgcaaaaat
     4021 attgcattct tcatgaatta aagttgcata taaggcatat tgaaagtaat agtactaaaa
     4081 cagcagttag cgaaaattaa tagaattata ttcgcaagac aattgtgaca ttaaattaaa
     4141 aaattgtaga atttttacta tcctctttaa cgccatgagc ctttataaaa aggttaaatt
     4201 agttttaaca ttcttttttt gagtaagagt ttacaattta tcaaaacctg tgttattata
     4261 ttcattagtt tcaatttatt agcatctaga gaaaaatcaa ttggcagtta cattgttgga
     4321 atttatgaag aaaaagaatc tacaacggag aataagttgc tgatcgcttt ccctaaaatt
     4381 gtatattttg ctgagcttat tttgacattt cgttgaagtt ttctctaatt cgcattcatt
     4441 ttaagtaaaa caatgagaaa taaaattaca aaaaatacaa attaaaatac aatttttagc
     4501 tataattata gacctaggct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     4561 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     4621 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     4681 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     4741 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     4801 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     4861 tcccttcggg aagcgtggcg ctttctcaat gctcacgctg taggtatctc agttcggtgt
     4921 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     4981 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     5041 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     5101 tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc
     5161 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     5221 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     5281 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     5341 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     5401 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     5461 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     5521 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     5581 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     5641 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     5701 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     5761 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     5821 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     5881 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     5941 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     6001 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     6061 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     6121 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     6181 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     6241 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     6301 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     6361 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     6421 catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct
     6481 ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa
     6541 acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga
     6601 gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact
     6661 atgcggcatc agagcagatt gtactgagag tgcacca
//
