LOCUS       sequence-146380-        6241 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 6241)
  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 6241)
  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: pNXRVc-HACre
FEATURES             Location/Qualifiers
     source          1..6241
                     /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"
     gene            2353..3472
                     /label=natMX6
                     /note="yeast selectable marker conferring nourseothricin 
                     resistance"
     promoter        2353..2696
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             2697..3269
                     /codon_start=1
                     /gene="Streptomyces noursei nat1"
                     /product="nourseothricin acetyltransferase"
                     /label=NrsR
                     /note="confers resistance to nourseothricin"
                     /translation="MGTTLDDTAYRYRTSVPGDAEAIEALDGSFTTDTVFRVTATGDGF
                     TLREVPVDPPLTKVFPDDESDDESDDGEDGDPDSRTFVAYGDDGDLAGFVVVSYSGWNR
                     RLTVEDIEVAPEHRGHGVGRALMGLATEFARERGAGHLWLEVTNVNAPAIHAYRRMGFT
                     LCGLDTALYDGTASDGEQALYMSMPCP"
     terminator      3275..3472
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     rep_origin      3527..4310
                     /label=ars1
                     /note="Schizosaccharomyces pombe autonomously replicating 
                     sequence ars1"
     primer_bind     complement(4321..4338)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4492..5080)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4572..4591)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5251..6042,6043..6111))
                     /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     5874..5893
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6112..6216)
                     /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 tctgtttagc ttgccttgtc cccgccgggt
     2341 cacccggcca gcgacatgga ggcccagaat accctccttg acagtcttga cgtgcgcagc
     2401 tcaggggcat gatgtgactg tcgcccgtac atttagccca tacatcccca tgtataatca
     2461 tttgcatcca tacattttga tggccgcacg gcgcgaagca aaaattacgg ctcctcgctg
     2521 cagacctgcg agcagggaaa cgctcccctc acagacgcgt tgaattgtcc ccacgccgcg
     2581 cccctgtaga gaaatataaa aggttaggat ttgccactga ggttcttctt tcatatactt
     2641 ccttttaaaa tcttgctagg atacagttct cacatcacat ccgaacataa acaaccatgg
     2701 gtaccactct tgacgacacg gcttaccggt accgcaccag tgtcccgggg gacgccgagg
     2761 ccatcgaggc actggatggg tccttcacca ccgacaccgt cttccgcgtc accgccaccg
     2821 gggacggctt caccctgcgg gaggtgccgg tggacccgcc cctgaccaag gtgttccccg
     2881 acgacgaatc ggacgacgaa tcggacgacg gggaggacgg cgacccggac tcccggacgt
     2941 tcgtcgcgta cggggacgac ggcgacctgg cgggcttcgt ggtcgtctcg tactccggct
     3001 ggaaccgccg gctgaccgtc gaggacatcg aggtcgcccc ggagcaccgg gggcacgggg
     3061 tcgggcgcgc gttgatgggg ctcgcgacgg agttcgcccg cgagcggggc gccgggcacc
     3121 tctggctgga ggtcaccaac gtcaacgcac cggcgatcca cgcgtaccgg cggatggggt
     3181 tcaccctctg cggcctggac accgccctgt acgacggcac cgcctcggac ggcgagcagg
     3241 cgctctacat gagcatgccc tgcccctaat cagtactgac aataaaaaga ttcttgtttt
     3301 caagaacttg tcatttgtat agttttttta tattgtagtt gttctatttt aatcaaatgt
     3361 tagcgtgatt tatatttttt ttcgcctcga catcatctgc ccagatgcga agttaagtgc
     3421 gcagaaagta atatcatgcg tcaatcgtat gtgaatgctg gtcgctatac tgctgtcgat
     3481 tcgatactaa cgccgccatc cagtttaaac gagctcgaat tctaggtagg cattttgttt
     3541 agttaaaggt acttaagtaa ttagtataaa cgaaccaatt ttataatcag gaagttaagt
     3601 gaatggtagc acatgtcgta aaaattgtga atttttattg aataatattt taaatacaag
     3661 cctttctaga ctaggtatac tcataaacat atatgagcaa aaggatagag gagattacat
     3721 tgcatcttct acaaatttat ttattgccct ttactgaaaa attaaataat gagtactaaa
     3781 tgataaaaag cgctcagtac aagaaagatt gcaaaaatat tgcattcttc atgaattaaa
     3841 gttgcatata aggcatattg aaagtaatag tactaaaaca gcagttagcg aaaattaata
     3901 gaattatatt cgcaagacaa ttgtgacatt aaattaaaaa attgtagaat ttttactatc
     3961 ctctttaacg ccatgagcct ttataaaaag gttaaattag ttttaacatt ctttttttga
     4021 gtaagagttt acaatttatc aaaacctgtg ttattatatt cattagtttc aatttattag
     4081 catctagaga aaaatcaatt ggcagttaca ttgttggaat ttatgaagaa aaagaatcta
     4141 caacggagaa taagttgctg atcgctttcc ctaaaattgt atattttgct gagcttattt
     4201 tgacatttcg ttgaagtttt ctctaattcg cattcatttt aagtaaaaca atgagaaata
     4261 aaattacaaa aaatacaaat taaaatacaa tttttagcta taattataga cctaggcttc
     4321 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     4381 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     4441 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     4501 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     4561 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     4621 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     4681 ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     4741 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     4801 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     4861 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     4921 ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     4981 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     5041 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     5101 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     5161 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     5221 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     5281 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     5341 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     5401 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     5461 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     5521 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     5581 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     5641 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     5701 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     5761 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     5821 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     5881 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     5941 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     6001 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     6061 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     6121 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     6181 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     6241 t
//
