LOCUS       sequence-146387-        6941 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 6941)
  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 6941)
  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: pNXRVj-HACre
FEATURES             Location/Qualifiers
     source          1..6941
                     /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             2656..3924
                     /codon_start=1
                     /product="aureobasidin A resistance protein from 
                     Schizosaccharomyces pombe (Hashida-Okado et al., 1998)"
                     /label=aur1-R
                     /translation="MSALSTLKKRLAACNRASQYKLETSLNPMPTFRLLRNTKWSWTHL
                     QYVFLAGNLIFACIVIESPGFWGKFGIACLLAIALTVPLTRQIFFPAIVIITWAILFYS
                     CRFIPERWRPPIWVRVLPTLENILYGSNLSSLLSKTTHSILDILAWVPYGVMHYSAPFI
                     ISFILFIFAPPGTLPVWARTFGYMNLFGVLIQMAFPCSPPWYENMYGLEPATYAVRGSP
                     GGLARIDALFGTSIYTDCFSNSPVVFGAFPSLHAGWAMLEALFLSHVFPRYRFCFYGYV
                     LWLCWCTMYLTHHYFVDLVGGMCLAIICFVFAQKLRLPQLQTGKILRWEYEFVIHGHGL
                     SEKTSNSLARTGSPYLLGRDSFTQNPNAVAFMSGLNNMELANTDHEWSVGSSSPEPLPS
                     PAADLIDRPASTTSSIFDASHLP"
     rep_origin      4227..5010
                     /label=ars1
                     /note="Schizosaccharomyces pombe autonomously replicating 
                     sequence ars1"
     primer_bind     complement(5021..5038)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5192..5780)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5272..5291)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5951..6742,6743..6811))
                     /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     6574..6593
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6812..6916)
                     /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 tctgtcagga aagctttttt gcctctgcaa
     2341 aagttccttt ctcgaattgg ttttttgagg aaaagcaagt taataaacta attatattat
     2401 atataattag caattttata aaaaaaataa aaaaatagcc ctgattgctg gcaactgtga
     2461 gctgaacatt ggttaatcgg tccatctttt tttaaatatt ttacatcgct acttttaagt
     2521 gcttgacact tgcatttaat agctactttc tttccttcat aaaaattcct tttttttcct
     2581 ttagttttcc ggttaattcc ttacgaaatt tttttcgtac gcttcccttt tttactctga
     2641 taattctttg aagcaatgtc tgctctttcg accttaaaaa agcgccttgc tgcgtgtaac
     2701 cgagcatccc aatacaagtt ggaaacaagc ttaaacccta tgcctacatt tcgtttgcta
     2761 cgcaatacga aatggtcatg gacacatttg caatatgtgt ttctagcagg taatttgatt
     2821 tttgcttgta ttgtcattga atctcctgga ttctggggga aatttggcat tgcctgtctt
     2881 ttggccattg cgttgaccgt tcctttaaca cgccaaattt tttttcctgc cattgttatc
     2941 atcacctggg caattttatt ttactcttgt aggtttattc cagaacgctg gcgtccaccc
     3001 atatgggttc gtgttttacc cacacttgaa aatattcttt atggctctaa tctttctagt
     3061 cttctctcga aaaccacgca tagcatcctt gatattttgg cctgggttcc atatggagtc
     3121 atgcattatt cggctccttt tatcatttca tttattcttt tcatctttgc acctcctgga
     3181 actcttccag tttgggctcg aacttttggt tatatgaatt tatttggtgt tcttatccaa
     3241 atggctttcc cctgttctcc tccttggtat gaaaatatgt atggtttaga acctgccacg
     3301 tatgcagtac gtggctctcc tggtggattg gcccgtattg atgctctctt cggcactagc
     3361 atttacactg attgtttttc taactctccg gttgtttttg gtgcctttcc atctcttcac
     3421 gctggatggg ccatgctgga agcacttttc ctttcgcatg tgtttcctcg ataccgcttc
     3481 tgcttttatg gatatgttct atggctttgc tggtgtacta tgtaccttac ccaccactac
     3541 tttgtagatt tggtcggcgg tatgtgttta gctattatat gcttcgtttt tgctcaaaag
     3601 ctacgcctcc cacagttgca aactggtaaa atccttcgtt gggaatacga gtttgttatc
     3661 cacggtcatg gtctttccga aaaaaccagc aactccttgg ctcgtaccgg cagcccatac
     3721 ttacttggaa gggattcttt tactcaaaac cctaatgcag tagccttcat gagtggtctt
     3781 aacaatatgg aacttgctaa caccgatcat gaatggtccg tgggttcatc atcacctgag
     3841 ccgttaccta gtcctgctgc tgatttgatt gatcgtcctg ccagtaccac ttcctccatc
     3901 tttgatgcaa gtcatcttcc ttaaatcaac gtgctttaag aatatatttc caaaagctac
     3961 atgatacatt gactagaatc ggtttgattc atagtggtat tggaatgatg ttgttcattg
     4021 tgttttttaa ctgttaatct gacatccatt gagtcattct ttacaatttg taaaattaat
     4081 ttgtatcact aattttgaag gaagctattt tggtattaat accgcttttg gtctccactt
     4141 ccttttcgaa actcttaaca gcgattaggc cgggtatctt ccagtgtgat gtataggtat
     4201 ttgtttaaac gagctcgaat tctaggtagg cattttgttt agttaaaggt acttaagtaa
     4261 ttagtataaa cgaaccaatt ttataatcag gaagttaagt gaatggtagc acatgtcgta
     4321 aaaattgtga atttttattg aataatattt taaatacaag cctttctaga ctaggtatac
     4381 tcataaacat atatgagcaa aaggatagag gagattacat tgcatcttct acaaatttat
     4441 ttattgccct ttactgaaaa attaaataat gagtactaaa tgataaaaag cgctcagtac
     4501 aagaaagatt gcaaaaatat tgcattcttc atgaattaaa gttgcatata aggcatattg
     4561 aaagtaatag tactaaaaca gcagttagcg aaaattaata gaattatatt cgcaagacaa
     4621 ttgtgacatt aaattaaaaa attgtagaat ttttactatc ctctttaacg ccatgagcct
     4681 ttataaaaag gttaaattag ttttaacatt ctttttttga gtaagagttt acaatttatc
     4741 aaaacctgtg ttattatatt cattagtttc aatttattag catctagaga aaaatcaatt
     4801 ggcagttaca ttgttggaat ttatgaagaa aaagaatcta caacggagaa taagttgctg
     4861 atcgctttcc ctaaaattgt atattttgct gagcttattt tgacatttcg ttgaagtttt
     4921 ctctaattcg cattcatttt aagtaaaaca atgagaaata aaattacaaa aaatacaaat
     4981 taaaatacaa tttttagcta taattataga cctaggcttc ctcgctcact gactcgctgc
     5041 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     5101 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     5161 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     5221 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     5281 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     5341 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcaatgc tcacgctgta
     5401 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     5461 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     5521 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     5581 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat
     5641 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     5701 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     5761 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     5821 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     5881 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     5941 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     6001 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     6061 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     6121 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     6181 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     6241 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     6301 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     6361 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     6421 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     6481 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     6541 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     6601 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     6661 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     6721 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     6781 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     6841 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     6901 aaataggggt tccgcgcaca tttccccgaa aagtgccacc t
//
