LOCUS       sequence-146381-        7705 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 7705)
  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 7705)
  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: pNXRVct-HACre
FEATURES             Location/Qualifiers
     source          1..7705
                     /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..4733
                     /codon_start=1
                     /gene="Streptomyces noursei nat1"
                     /product="nourseothricin acetyltransferase"
                     /label=NrsR
                     /note="confers resistance to nourseothricin"
                     /translation="MGTTLDDTAYRYRTSVPGDAEAIEALDGSFTTDTVFRVTATGDGF
                     TLREVPVDPPLTKVFPDDESDDESDDGEDGDPDSRTFVAYGDDGDLAGFVVVSYSGWNR
                     RLTVEDIEVAPEHRGHGVGRALMGLATEFARERGAGHLWLEVTNVNAPAIHAYRRMGFT
                     LCGLDTALYDGTASDGEQALYMSMPCP"
     terminator      4739..4936
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     rep_origin      4991..5774
                     /label=ars1
                     /note="Schizosaccharomyces pombe autonomously replicating 
                     sequence ars1"
     primer_bind     complement(5785..5802)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5956..6544)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6036..6055)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6715..7506,7507..7575))
                     /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     7338..7357
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7576..7680)
                     /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 ggtgggagga ttggggacag ctttcgggga
     3841 cggccgtgcc gccccagggt gccgagcccc agagcaacgc gggcccacga ccccatatcg
     3901 gggacacgtt atttaccctg tttcgggccc ccgagttgct ggcccccaac ggcgacctgt
     3961 ataacgtgtt tgcctgggcc ttggacgtct tggccaaacg cctccgtccc atgcacgtct
     4021 ttatcctgga ttacgaccaa tcgcccgccg gctgccggga cgccctgctg caacttacct
     4081 ccgggatggt ccagacccac gtcaccaccc caggctccat accgacgatc tgcgacctgg
     4141 cgcgcacgtt tgcccgggcc atgggtacca ctcttgacga cacggcttac cggtaccgca
     4201 ccagtgtccc gggggacgcc gaggccatcg aggcactgga tgggtccttc accaccgaca
     4261 ccgtcttccg cgtcaccgcc accggggacg gcttcaccct gcgggaggtg ccggtggacc
     4321 cgcccctgac caaggtgttc cccgacgacg aatcggacga cgaatcggac gacggggagg
     4381 acggcgaccc ggactcccgg acgttcgtcg cgtacgggga cgacggcgac ctggcgggct
     4441 tcgtggtcgt ctcgtactcc ggctggaacc gccggctgac cgtcgaggac atcgaggtcg
     4501 ccccggagca ccgggggcac ggggtcgggc gcgcgttgat ggggctcgcg acggagttcg
     4561 cccgcgagcg gggcgccggg cacctctggc tggaggtcac caacgtcaac gcaccggcga
     4621 tccacgcgta ccggcggatg gggttcaccc tctgcggcct ggacaccgcc ctgtacgacg
     4681 gcaccgcctc ggacggcgag caggcgctct acatgagcat gccctgcccc taatcagtac
     4741 tgacaataaa aagattcttg ttttcaagaa cttgtcattt gtatagtttt tttatattgt
     4801 agttgttcta ttttaatcaa atgttagcgt gatttatatt ttttttcgcc tcgacatcat
     4861 ctgcccagat gcgaagttaa gtgcgcagaa agtaatatca tgcgtcaatc gtatgtgaat
     4921 gctggtcgct atactgctgt cgattcgata ctaacgccgc catccagttt aaacgagctc
     4981 gaattctagg taggcatttt gtttagttaa aggtacttaa gtaattagta taaacgaacc
     5041 aattttataa tcaggaagtt aagtgaatgg tagcacatgt cgtaaaaatt gtgaattttt
     5101 attgaataat attttaaata caagcctttc tagactaggt atactcataa acatatatga
     5161 gcaaaaggat agaggagatt acattgcatc ttctacaaat ttatttattg ccctttactg
     5221 aaaaattaaa taatgagtac taaatgataa aaagcgctca gtacaagaaa gattgcaaaa
     5281 atattgcatt cttcatgaat taaagttgca tataaggcat attgaaagta atagtactaa
     5341 aacagcagtt agcgaaaatt aatagaatta tattcgcaag acaattgtga cattaaatta
     5401 aaaaattgta gaatttttac tatcctcttt aacgccatga gcctttataa aaaggttaaa
     5461 ttagttttaa cattcttttt ttgagtaaga gtttacaatt tatcaaaacc tgtgttatta
     5521 tattcattag tttcaattta ttagcatcta gagaaaaatc aattggcagt tacattgttg
     5581 gaatttatga agaaaaagaa tctacaacgg agaataagtt gctgatcgct ttccctaaaa
     5641 ttgtatattt tgctgagctt attttgacat ttcgttgaag ttttctctaa ttcgcattca
     5701 ttttaagtaa aacaatgaga aataaaatta caaaaaatac aaattaaaat acaattttta
     5761 gctataatta tagacctagg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     5821 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     5881 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     5941 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     6001 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     6061 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     6121 tctcccttcg ggaagcgtgg cgctttctca atgctcacgc tgtaggtatc tcagttcggt
     6181 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     6241 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     6301 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     6361 cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct
     6421 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     6481 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     6541 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     6601 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     6661 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     6721 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     6781 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     6841 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     6901 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     6961 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     7021 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     7081 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     7141 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     7201 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     7261 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     7321 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     7381 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     7441 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     7501 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     7561 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     7621 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     7681 cacatttccc cgaaaagtgc cacct
//
