LOCUS       sequence-322938-        6906 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Constitutive expression vector for Cre-recombinase containing URA3 
            selection marker..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6906)
  AUTHORS   MacDonald C, Piper RC
  TITLE     Puromycin and Methotrexate Resistance Cassettes and Optimized 
            cre-recombinase Expression Plasmids for use in Yeast.
  JOURNAL   Yeast. 2015 Feb 16. doi: 10.1002/yea.3069.
  PUBMED    25688547
REFERENCE   2  (bases 1 to 6906)
  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: "Yeast. 2015
            Feb 16. doi: 10.1002/yea.3069."
COMMENT     Sequence Label: pPL5071_TEF1*-Cre_URA3
FEATURES             Location/Qualifiers
     source          1..6906
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..19
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     CDS             553..1584
                     /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="MSNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKML
                     LSVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGL
                     PRPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAF
                     LGIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVE
                     RWISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQ
                     RYLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLE
                     DGD"
     terminator      1902..2149
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(1918..1936)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(2167..2186)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2168..2186)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2196..2213)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2196..2212)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2205..2227)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2353..2808
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2440..2459)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2650..2671
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2939..3742)
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     promoter        complement(3743..3958)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(3982..4001)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4101..4123
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4161..4179)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    4220..4723
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        4760..4864
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4865..4933,4934..5725)
                     /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     complement(5083..5102)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5896..6484
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6385..6404
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6638..6655
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6772..6793
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6808..6813,6814..6831,6832..6838)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6846..6862
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     6851..6873
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6870..6886
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6870..6886
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     join(6905..6906,1..19)
                     /label=T3
                     /note="T3 promoter, forward primer"
ORIGIN
        1 aattaaccct cactaaaggg aacaaaagct ggatagcttc aaaatgtctc tactcctttt
       61 ttactcttcc agattttctc ggactccgcg caccgccgta ccacttcaaa acacccaagc
      121 acagcatact aaattccccc tcctccttcc tctagggtgc cgttaattac ccgtactaaa
      181 ggtttggaaa aggaaaaaga gaccgcctcg tccctttttc ttcgtcggag aaggcaataa
      241 aaatttttat cacgtttctt tctcttgaaa actttttttt cgattttgtt ctctttcgac
      301 gacctcccat tgatatttga gttaacaaac ggtcttcaat ttctcaagtt tcagcttcat
      361 ttttcctgtt ctattacaac tttttttact tcttgctcat tggaaagaaa gcatagcaat
      421 ctaatctaag ttttaattac aaaaactagt ggatcccccg ggctgcagga attcgatatc
      481 aagcttatcg ataccgtcga ggggcagagc cgatcctgta cactttactt aaaaccatta
      541 tctgagtgtt aaatgtccaa tttactgacc gtacaccaaa atttgcctgc attaccggtc
      601 gatgcaacga gtgatgaggt tcgcaagaac ctgatggaca tgttcaggga tcgccaggcg
      661 ttttctgagc atacctggaa aatgcttctg tccgtttgcc ggtcgtgggc ggcatggtgc
      721 aagttgaata accggaaatg gtttcccgca gaacctgaag atgttcgcga ttatcttcta
      781 tatcttcagg cgcgcggtct ggcagtaaaa actatccagc aacatttggg ccagctaaac
      841 atgcttcatc gtcggtccgg gctgccacga ccaagtgaca gcaatgctgt ttcactggtt
      901 atgcggcgga tccgaaaaga aaacgttgat gccggtgaac gtgcaaaaca ggctctagcg
      961 ttcgaacgca ctgatttcga ccaggttcgt tcactcatgg aaaatagcga tcgctgccag
     1021 gatatacgta atctggcatt tctggggatt gcttataaca ccctgttacg tatagccgaa
     1081 attgccagga tcagggttaa agatatctca cgtactgacg gtgggagaat gttaatccat
     1141 attggcagaa cgaaaacgct ggttagcacc gcaggtgtag agaaggcact tagcctgggg
     1201 gtaactaaac tggtcgagcg atggatttcc gtctctggtg tagctgatga tccgaataac
     1261 tacctgtttt gccgggtcag aaaaaatggt gttgccgcgc catctgccac cagccagcta
     1321 tcaactcgcg ccctggaagg gatttttgaa gcaactcatc gattgattta cggcgctaag
     1381 gatgactctg gtcagagata cctggcctgg tctggacaca gtgcccgtgt cggagccgcg
     1441 cgagatatgg cccgcgctgg agtttcaata ccggagatca tgcaagctgg tggctggacc
     1501 aatgtaaata ttgtcatgaa ctatatccgt aacctggata gtgaaacagg ggcaatggtg
     1561 cgcctgctgg aagatggcga ttagccatta acgcgtaaat gattgctata attatttgat
     1621 atttatggtg acatatgaga aaggatttca acatcgacgg aaaatatgta gtgctgtctg
     1681 taagcactaa tattcagtcg ccagccgtca ttgtcactgt aaagctgagc gatagaatgc
     1741 ctgatattga ctcaatatcc gttgcgtttc ctgtcaaaag tatgcgtagt gctgaacatt
     1801 tcgtgatgaa tgccaccgag gaagaagcac ggcgcggttt tgctaaagtg atgtctgagt
     1861 ttggcgaact cttgggtaag gttggaattg tcgacctcga gtcatgtaat tagttatgtc
     1921 acgcttacat tcacgccctc cccccacatc cgctctaacc gaaaaggaag gagttagaca
     1981 acctgaagtc taggtcccta tttatttttt tatagttatg ttagtattaa gaacgttatt
     2041 tatatttcaa atttttcttt tttttctgta cagacgcgtg tacgcatgta acattatact
     2101 gaaaaccttg cttgagaagg ttttgggacg ctcgaaggct ttaatttgcg gccggtaccc
     2161 aattcgccct atagtgagtc gtattacgcg cgctcactgg ccgtcgtttt acaacgtcgt
     2221 gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc
     2281 agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg
     2341 aatggcgaat ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     2401 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     2461 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     2521 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
     2581 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     2641 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
     2701 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     2761 aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttcc tgatgcggta
     2821 ttttctcctt acgcatctgt gcggtatttc acaccgcata gggtaataac tgatataatt
     2881 aaattgaagc tctaatttgt gagtttagta tacatgcatt tacttataat acagtttttt
     2941 agttttgctg gccgcatctt ctcaaatatg cttcccagcc tgcttttctg taacgttcac
     3001 cctctacctt agcatccctt ccctttgcaa atagtcctct tccaacaata ataatgtcag
     3061 atcctgtaga gaccacatca tccacggttc tatactgttg acccaatgcg tctcccttgt
     3121 catctaaacc cacaccgggt gtcataatca accaatcgta accttcatct cttccaccca
     3181 tgtctctttg agcaataaag ccgataacaa aatctttgtc gctcttcgca atgtcaacag
     3241 tacccttagt atattctcca gtagataggg agcccttgca tgacaattct gctaacatca
     3301 aaaggcctct aggttccttt gttacttctt ctgccgcctg cttcaaaccg ctaacaatac
     3361 ctgggcccac cacaccgtgt gcattcgtaa tgtctgccca ttctgctatt ctgtatacac
     3421 ccgcagagta ctgcaatttg actgtattac caatgtcagc aaattttctg tcttcgaaga
     3481 gtaaaaaatt gtacttggcg gataatgcct ttagcggctt aactgtgccc tccatggaaa
     3541 aatcagtcaa gatatccaca tgtgttttta gtaaacaaat tttgggacct aatgcttcaa
     3601 ctaactccag taattccttg gtggtacgaa catccaatga agcacacaag tttgtttgct
     3661 tttcgtgcat gatattaaat agcttggcag caacaggact aggatgagta gcagcacgtt
     3721 ccttatatgt agctttcgac atgatttatc ttcgtttcct gcaggttttt gttctgtgca
     3781 gttgggttaa gaatactggg caatttcatg tttcttcaac actacatatg cgtatatata
     3841 ccaatctaag tctgtgctcc ttccttcgtt cttccttctg ttcggagatt accgaatcaa
     3901 aaaaatttca aggaaaccga aatcaaaaaa aagaataaaa aaaaaatgat gaattgaaaa
     3961 ggtggtatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg
     4021 acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta
     4081 cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc
     4141 gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat
     4201 aataatggtt tcttagacgg atcgcttgcc tgtaacttac acgcgcctcg tatcttttaa
     4261 tgatggaata atttgggaat ttactctgtg tttatttatt tttatgtttt gtatttggat
     4321 tttagaaagt aaataaagaa ggtagaagag ttacggaatg aagaaaaaaa aataaacaaa
     4381 ggtttaaaaa atttcaacaa aaagcgtact ttacatatat atttattaga caagaaaagc
     4441 agattaaata gatatacatt cgattaacga taagtaaaat gtaaaatcac aggattttcg
     4501 tgtgtggtct tctacacaga caagatgaaa caattcggca ttaatacctg agagcaggaa
     4561 gagcaagata aaaggtagta tttgttggcg atccccctag agtcttttac atcttcggaa
     4621 aacaaaaact attttttctt taatttcttt ttttactttc tatttttaat ttatatattt
     4681 atattaaaaa atttaaatta taattatttt tatagcacgt gatgaaaagg acccaggtgg
     4741 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     4801 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     4861 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     4921 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     4981 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     5041 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     5101 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     5161 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     5221 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     5281 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     5341 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     5401 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     5461 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     5521 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     5581 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     5641 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     5701 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     5761 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     5821 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     5881 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     5941 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     6001 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     6061 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     6121 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     6181 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     6241 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     6301 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     6361 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     6421 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     6481 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     6541 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     6601 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     6661 ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag
     6721 ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag
     6781 ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg
     6841 tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa
     6901 gcgcgc
//
