LOCUS       sequence-146384-        6828 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 6828)
  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 6828)
  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: pNXRVd-HACre
FEATURES             Location/Qualifiers
     source          1..6828
                     /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             2859..3653
                     /codon_start=1
                     /gene="Schizosaccharomyces pombe ura4"
                     /product="orotidine 5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=S. pombe ura4
                     /translation="MDARVFQSYSARAEGMKNPIAKELLALMEEKQSNLSVAVDLTKKS
                     EILELVDKIGPYVCVIKTHIDVVEDFDQDMVEKLVALGKKHRFLIFEDRKFADIGNTVK
                     LQYASGVYKIASWAHITNCHTVPGEGIIQGLKEVGLPLGRGLLLLAEMSSKGSLATGSY
                     TEKTLEWFEKHTDFCFGFIAGRRFPNLQSDYITMSPGIGLDVKGDGLGQQYRTPEEVIV
                     NCGSDIIIVGRGVYGAGRNPVVEAKRYREAGWKAYQQRLSQH"
     rep_origin      4114..4897
                     /label=ars1
                     /note="Schizosaccharomyces pombe autonomously replicating 
                     sequence ars1"
     primer_bind     complement(4908..4925)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5079..5667)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5159..5178)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5838..6629,6630..6698))
                     /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     6461..6480
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6699..6803)
                     /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 tctgagctcg atatcaagct tagctacaaa
     2341 tcccactggc tatatgtatg catttgtgtt aaaaaagttt gtatagatta tttaatctac
     2401 tcagcattct ttctctaaat aggaatttgt tacttaatgg agaaaaaaat gtttcgattt
     2461 acctagtgta tttgtttgta tactcacgtt taatttcaaa catccattct atcttgtgta
     2521 atttttggca tggtgaaaaa gataatcagc cttataatct ttacaaaagt aagaaattct
     2581 gtaaataagc cttaatgccc ttgctttaaa ttaaaatggt tctttttcat gataatgttt
     2641 gcactttgtg aatatatttt agatagttct gtgaggtata attaagatgt tttagagact
     2701 tatacaattt tgtctttata aattcttaat tgattttacc atcccagttt aactatgctt
     2761 cgtcggcatc tctgcacatg tcgtgttttc ttaccgtatt gtcctaccaa gaacctcttt
     2821 tttgcttgga tcgaaattaa aggtttaaaa gcaaagttat ggatgctaga gtatttcaaa
     2881 gctattcagc tagagctgag gggatgaaaa atcccattgc caaggaattg ttggctttga
     2941 tggaagaaaa gcaaagcaac ttgtcagtcg cggtcgattt gacgaagaaa tccgaaatct
     3001 tagaattggt agataaaatt ggaccctatg tctgtgttat caagacacat attgacgttg
     3061 tcgaggattt cgaccaggat atggtagaaa aactggtggc cttaggtaaa aagcatcgtt
     3121 ttcttatctt tgaggatcgc aaattcgcag acattggaaa taccgtcaag ctacaatatg
     3181 catctggtgt gtacaaaatt gcttcttggg ctcatatcac aaattgccat acagtgccag
     3241 gcgagggtat tatacaaggc ctcaaagaag ttggtttacc tttgggacgt ggtctcttgc
     3301 ttttggctga aatgtcttcc aaaggctctt tggctactgg ttcctacaca gagaaaacct
     3361 tagaatggtt tgagaagcat accgattttt gctttggctt tatagctggt cgtcgatttc
     3421 ctaaccttca aagcgactac ataactatgt cccctggtat cggcttggat gttaaaggag
     3481 acgggctggg acagcaatat cgtactcctg aagaagtgat tgtaaactgc ggtagcgata
     3541 tcatcattgt tggtcgtgga gtctatggag ctggtcgtaa tcctgttgtc gaagccaaga
     3601 gatatagaga agctggttgg aaggcatatc agcaaagact ttctcagcat taaaaaaaga
     3661 ctaatgtaaa atttttttgg ttggttattg aaaaagtcga tgccttgttt gcgtttgttt
     3721 tcctaggcgt tttatgtcag aaggcattta gaattagtat acaagtactc tttggtaaaa
     3781 ttttatgtag cgactaaaat attaactatt atagataaac accttgggaa taaaaagtaa
     3841 tttgctatag taatttatta aacatgctcc tacaacatta ccacaatctt ttctcttgga
     3901 ttgacattga ataagaaaag agtgaatttt tttagacttg taatgataac tatgtacaaa
     3961 gccaatgaaa gatgtatgta gatgaatgta aaataccatg tagacaaaca agataaaact
     4021 tggttataaa cattggtgtt ggaacagaat aaattagatg tcaaaaagtt tcgtcaatat
     4081 cacaagcttg tttaaacgag ctcgaattct aggtaggcat tttgtttagt taaaggtact
     4141 taagtaatta gtataaacga accaatttta taatcaggaa gttaagtgaa tggtagcaca
     4201 tgtcgtaaaa attgtgaatt tttattgaat aatattttaa atacaagcct ttctagacta
     4261 ggtatactca taaacatata tgagcaaaag gatagaggag attacattgc atcttctaca
     4321 aatttattta ttgcccttta ctgaaaaatt aaataatgag tactaaatga taaaaagcgc
     4381 tcagtacaag aaagattgca aaaatattgc attcttcatg aattaaagtt gcatataagg
     4441 catattgaaa gtaatagtac taaaacagca gttagcgaaa attaatagaa ttatattcgc
     4501 aagacaattg tgacattaaa ttaaaaaatt gtagaatttt tactatcctc tttaacgcca
     4561 tgagccttta taaaaaggtt aaattagttt taacattctt tttttgagta agagtttaca
     4621 atttatcaaa acctgtgtta ttatattcat tagtttcaat ttattagcat ctagagaaaa
     4681 atcaattggc agttacattg ttggaattta tgaagaaaaa gaatctacaa cggagaataa
     4741 gttgctgatc gctttcccta aaattgtata ttttgctgag cttattttga catttcgttg
     4801 aagttttctc taattcgcat tcattttaag taaaacaatg agaaataaaa ttacaaaaaa
     4861 tacaaattaa aatacaattt ttagctataa ttatagacct aggcttcctc gctcactgac
     4921 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     4981 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     5041 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     5101 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     5161 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     5221 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcaatgctca
     5281 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     5341 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     5401 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     5461 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg
     5521 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     5581 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     5641 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     5701 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     5761 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     5821 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     5881 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     5941 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     6001 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     6061 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     6121 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     6181 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     6241 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     6301 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     6361 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     6421 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     6481 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     6541 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     6601 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     6661 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     6721 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     6781 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacct
//
