LOCUS       sequence-153388-        7942 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 7942)
  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 7942)
  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: pNXRVat-HACre
FEATURES             Location/Qualifiers
     source          1..7942
                     /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"
     CDS             4161..4970
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(4228..4247)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      4976..5173
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     rep_origin      5228..6011
                     /label=ars1
                     /note="Schizosaccharomyces pombe autonomously replicating 
                     sequence ars1"
     primer_bind     complement(6022..6039)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6193..6781)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6273..6292)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6952..7743,7744..7812))
                     /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     7575..7594
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7813..7917)
                     /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 tctcgtatgc ctacaacaac taagaaaatg
     2341 gctatcatgc ggaagctggt gagaagaaca gcatcgggac aagggaagga agaacaaaga
     2401 caaagaaaac aaaagaaagc aattgaaaac aaaacaaaac aattttcatt ccttctctta
     2461 tcattccttt tcttttcttt tctctcattc aacgcactcc atcgtatccg tattcctctt
     2521 attttttctc tttctctata tccatttctt tctctctagg tgtgtcctct ctctctcttc
     2581 aatttctcta ctccgcattc caacgcatcc ttcccccaac ctcccatttc ctccttacgg
     2641 cccgatagcg atcgtctttc cctcgctatc actcgctacc ggcccctcct ctgcaccgta
     2701 acctcctacg tatttaccat atcataaagt tttttccgac gcttatcgct gaccccctgt
     2761 cgccctccta ttggcttccg gattatcttc ttgtccataa ggtgatccat gcttcctgaa
     2821 gattcccgaa atgtgtccac tttggcgggg aatcattcca tccacttctt tctctctcgc
     2881 tttcctcatt cggcgctccc cttccgcgtc tcattggtct tccgctccgt ttttgctttg
     2941 ccgatgttac ttggggagag gtgcgataat cctttcgcaa aaactcggtt tgacgcctcc
     3001 catggtataa atagtgggtg gtggacaggt gccttcgctt ttcttttctt taagcaagag
     3061 aagcttccat ggaaactagt gcgttcgacc aggctgcgcg ttctcgcggc catagcaacc
     3121 gacgtacggc gttgcgccct cgccggcagc aagaagccac ggaagtccgc ctggagcaga
     3181 aaatgcccac gctactgcgg gtttatatag acggtcctca cgggatgggg aaaaccacca
     3241 ccacgcaact gctggtggcc ctgggttcgc gcgacgatat cgtctacgta cccgagccga
     3301 tgacttactg gcaggtgctg ggggcttccg agacaatcgc gaacatctac accacacaac
     3361 accgcctcga ccagggtgag atatcggccg gggacgcggc ggtggtaatg acaagcgccc
     3421 agataacaat gggcatgcct tatgccgtga ccgacgccgt tctggctcct catatcgggg
     3481 gggaggctgg gagctcacat gccccgcccc cggccctcac cctcatcttc gaccgccatc
     3541 ccatcgccgc cctcctgtgc tacccggccg cgcgatacct tatgggcagc atgacccccc
     3601 aggccgtgct ggcgttcgtg gccctcatcc cgccgacctt gcccggcaca aacatcgtgt
     3661 tgggggccct tccggaggac agacacatcg accgcctggc caaacgccag cgccccggcg
     3721 agcggcttga cctggctatg ctggccgcga ttcgccgcgt ttacgggctg cttgccaata
     3781 cggtgcggta tctgcagggc ggcgggtcgt ggcgggagga ttggggacag ctttcgggga
     3841 cggccgtgcc gccccagggt gccgagcccc agagcaacgc gggcccacga ccccatatcg
     3901 gggacacgtt atttaccctg tttcgggccc ccgagttgct ggcccccaac ggcgacctgt
     3961 acaacgtgtt tgcctgggcc ttggacgtct tggccaaacg cctccgtccc atgcacgtct
     4021 ttatcctgga ttacgaccaa tcgcccgccg gctgccggga cgccctgctg caacttacct
     4081 ccgggatggt ccagacccac gtcaccaccc ccggctccat accgacgatc tgcgacctgg
     4141 cgcgcacgtt tgcccgggcc atgggtaagg aaaagactca cgtttcgagg ccgcgattaa
     4201 attccaacat ggatgctgat ttatatgggt ataaatgggc tcgcgataat gtcgggcaat
     4261 caggtgcgac aatctatcga ttgtatggga agcccgatgc gccagagttg tttctgaaac
     4321 atggcaaagg tagcgttgcc aatgatgtta cagatgagat ggtcagacta aactggctga
     4381 cggaatttat gcctcttccg accatcaagc attttatccg tactcctgat gatgcatggt
     4441 tactcaccac tgcgatcccc ggcaaaacag cattccaggt attagaagaa tatcctgatt
     4501 caggtgaaaa tattgttgat gcgctggcag tgttcctgcg ccggttgcat tcgattcctg
     4561 tttgtaattg tccttttaac agcgatcgcg tatttcgtct cgctcaggcg caatcacgaa
     4621 tgaataacgg tttggttgat gcgagtgatt ttgatgacga gcgtaatggc tggcctgttg
     4681 aacaagtctg gaaagaaatg cataagcttt tgccattctc accggattca gtcgtcactc
     4741 atggtgattt ctcacttgat aaccttattt ttgacgaggg gaaattaata ggttgtattg
     4801 atgttggacg agtcggaatc gcagaccgat accaggatct tgccatccta tggaactgcc
     4861 tcggtgagtt ttctccttca ttacagaaac ggctttttca aaaatatggt attgataatc
     4921 ctgatatgaa taaattgcag tttcatttga tgctcgatga gtttttctaa tcagtactga
     4981 caataaaaag attcttgttt tcaagaactt gtcatttgta tagttttttt atattgtagt
     5041 tgttctattt taatcaaatg ttagcgtgat ttatattttt tttcgcctcg acatcatctg
     5101 cccagatgcg aagttaagtg cgcagaaagt aatatcatgc gtcaatcgta tgtgaatgct
     5161 ggtcgctata ctgctgtcga ttcgatacta acgccgccat ccagtttaaa cgagctcgaa
     5221 ttctaggtag gcattttgtt tagttaaagg tacttaagta attagtataa acgaaccaat
     5281 tttataatca ggaagttaag tgaatggtag cacatgtcgt aaaaattgtg aatttttatt
     5341 gaataatatt ttaaatacaa gcctttctag actaggtata ctcataaaca tatatgagca
     5401 aaaggataga ggagattaca ttgcatcttc tacaaattta tttattgccc tttactgaaa
     5461 aattaaataa tgagtactaa atgataaaaa gcgctcagta caagaaagat tgcaaaaata
     5521 ttgcattctt catgaattaa agttgcatat aaggcatatt gaaagtaata gtactaaaac
     5581 agcagttagc gaaaattaat agaattatat tcgcaagaca attgtgacat taaattaaaa
     5641 aattgtagaa tttttactat cctctttaac gccatgagcc tttataaaaa ggttaaatta
     5701 gttttaacat tctttttttg agtaagagtt tacaatttat caaaacctgt gttattatat
     5761 tcattagttt caatttatta gcatctagag aaaaatcaat tggcagttac attgttggaa
     5821 tttatgaaga aaaagaatct acaacggaga ataagttgct gatcgctttc cctaaaattg
     5881 tatattttgc tgagcttatt ttgacatttc gttgaagttt tctctaattc gcattcattt
     5941 taagtaaaac aatgagaaat aaaattacaa aaaatacaaa ttaaaataca atttttagct
     6001 ataattatag acctaggctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     6061 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     6121 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     6181 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     6241 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     6301 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     6361 cccttcggga agcgtggcgc tttctcaatg ctcacgctgt aggtatctca gttcggtgta
     6421 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     6481 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     6541 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     6601 gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct
     6661 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     6721 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     6781 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     6841 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     6901 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     6961 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     7021 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     7081 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     7141 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     7201 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     7261 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     7321 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     7381 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     7441 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     7501 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     7561 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     7621 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     7681 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     7741 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     7801 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     7861 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     7921 atttccccga aaagtgccac ct
//
