LOCUS       sequence-73691-e        8873 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8873)
  TITLE     Series-12:  S cerevisiae Expression Vectors
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8873)
  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: 
            "Unpublished"
COMMENT     Sequence Label: Gal1/10 Ura S. cerevisiae expression vector
            (1...
FEATURES             Location/Qualifiers
     source          1..8873
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(704..722)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        790..894
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(895..963,964..1755)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1113..1132)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1926..2514
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2415..2434
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2668..2685
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2700..2840
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2766..2785)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2926..2948
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2942..3133)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     promoter        4551..5215
                     /gene="GAL1 and GAL10 genes of S. cerevisiae"
                     /label=GAL1,10 promoter
                     /note="divergent inducible promoter, regulated by Gal4"
     protein_bind    4767..4884
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     5179..5202
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             complement(6266..7069)
                     /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(7070..7285)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     rep_origin      join(7877..8873,1..346)
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(join(8840..8873,1..14))
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
ORIGIN
        1 ggaataggaa cttcaaagcg tttccgaaaa cgagcgcttc cgaaaatgca acgcgagctg
       61 cgcacataca gctcactgtt cacgtcgcac ctatatctgc gtgttgcctg tatatatata
      121 tacatgagaa gaacggcata gtgcgtgttt atgcttaaat gcgtacttat atgcgtctat
      181 ttatgtagga tgaaaggtag tctagtacct cctgtgatat tatcccattc catgcggggt
      241 atcgtatgct tccttcagca ctacccttta gctgttctat atgctgccac tcctcaattg
      301 gattagtctc atccttcaat gctatcattt cctttgatat tggatcatat gcatagtacc
      361 gagaaactag tgcgaagtag tgatcaggta ttgctgttat ctgatgagta tacgttgtcc
      421 tggccacggc agaagcacgc ttatcgctcc aatttcccac aacattagtc aactccgtta
      481 ggcccttcat tgaaagaaat gaggtcatca aatgtcttcc aatgtgagat tttgggccat
      541 tttttatagc aaagattgaa taaggcgcat ttttcttcaa agctttattg tacgatctga
      601 ctaagttatc ttttaataat tggtattcct gtttattgct tgaagaattg ccggtcctat
      661 ttactcgttt taggactggt tcagaattct tgaagacgaa agggcctcgt gatacgccta
      721 tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg
      781 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
      841 ctcatgagac aataaccctg ataaatgctt caataataat gaaaaaggaa gagtatgagt
      901 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
      961 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     1021 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     1081 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtgtt
     1141 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     1201 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     1261 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     1321 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     1381 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgca
     1441 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     1501 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     1561 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     1621 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     1681 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     1741 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     1801 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     1861 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     1921 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     1981 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     2041 ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac
     2101 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     2161 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     2221 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     2281 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     2341 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     2401 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     2461 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     2521 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     2581 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     2641 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     2701 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat atggtgcact
     2761 ctcagtacaa tctgctctga tgccgcatag ttaagccagt atacactccg ctatcgctac
     2821 gtgactgggt catggctgcg ccccgacacc cgccaacacc cgctgacgcg ccctgacggg
     2881 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt
     2941 gtcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca gctgcggtaa agctcatcag
     3001 cgtggtcgtg aagcgattca cagatgtctg cctgttcatc cgcgtccagc tcgttgagtt
     3061 tctccagaag cgttaatgtc tggcttctga taaagcgggc catgttaagg gcggtttttt
     3121 cctgtttggt cacttgatgc ctccgtgtaa gggggaattt ctgttcatgg gggtaatgat
     3181 accgatgaaa cgagagagga tgctcacgat acgggttact gatgatgaac atgcccggtt
     3241 actggaacgt tgtgagggta aacaactggc ggtatggatg cggcgggacc agagaaaaat
     3301 cactcagggt caatgccagc gcttcgttaa tacagatgta ggtgttccac agggtagcca
     3361 gcagcatcct gcgatgcaga tccggaacat aatggtgcag ggcgctgact tccgcgtttc
     3421 cagactttac gaaacacgga aaccgaagac cattcatgtt gttgctcagg tcgcagacgt
     3481 tttgcagcag cagtcgcttc acgttcgctc gcgtatcggt gattcattct gctaaccagt
     3541 aaggcaaccc cgccagccta gccgggtcct caacgacagg agcacgatca tgcgcacccg
     3601 tggccaggac ccaacgctgc ccgagatgcg ccgcgtgcgg ctgctggaga tggcggacgc
     3661 gatggatatg ttctgccaag ggttggtttg cgcattcaca gttctccgca agaattgatt
     3721 ggctccaatt cttggagtgg tgaatccgtt agcgaggtgc cgccggcttc cattcaggtc
     3781 gaggtggccc ggctccatgc accgcgacgc aacgcgggga ggcagacaag gtatagggcg
     3841 gcgcctacaa tccatgccaa cccgttccat gtgctcgccg aggcggcata aatcgccgtg
     3901 acgatcagcg gtccagtgat cgaagttagg ctggtaagag ccgcgagcga tccttgaagc
     3961 tgtccctgat ggtcgtcatc tacctgcctg gacagcatgg cctgcaacgc gggcatcccg
     4021 atgccgccgg aagcgagaag aatcataatg gggaaggcca tccagcctcg cgtcgcgaac
     4081 gccagcaaga cgtagcccag cgcgtcggcc gccatgccgg cgataatggc ctgcttctcg
     4141 ccgaaacgtt tggtggcggg accagtgacg aaggcttgag cgagggcgtg caagattccg
     4201 aataccgcaa gcgacaggcc gatcatcgtc gcgctccagc gaaagcggtc ctcgccgaaa
     4261 atgacccaga gcgctgccgg cacctgtcct acgagttgca tgataaagaa gacagtcata
     4321 agtgcggcga cgatagtcat gccccgcgcc caccggaagg agctgactgg gttgaaggct
     4381 ctcaagggca tcggtcgacc aattcgacag gttatcagca acaacacagt catatccatt
     4441 ctcaattagc tctaccacag tgtgtgaacc aatgtatcca gcaccacctg taaccaaaac
     4501 aattttagaa gtactttcac tttgtaactg agctgtcatt tatattgaat tttcaaaaat
     4561 tcttactttt tttttggatg gacgcaaaga agtttaataa tcatattaca tggcattacc
     4621 accatataca tatccatata catatccata tctaatctta cttatatgtt gtggaaatgt
     4681 aaagagcccc attatcttag cctaaaaaaa ccttctcttt ggaactttca gtaatacgct
     4741 taactgctca ttgctatatt gaagtacgga ttagaagccg ccgagcgggt gacagccctc
     4801 cgaaggaaga ctctcctccg tgcgtcctcg tcttcaccgg tcgcgttcct gaaacgcaga
     4861 tgtgcctcgc gccgcactgc tccgaacaat aaagattcta caatactagc ttttatggtt
     4921 atgaagagga aaaattggca gtaacctggc cccacaaacc ttcaaatgaa cgaatcaaat
     4981 taacaaccat aggatgataa tgcgattagt tttttagcct tatttctggg gtaattaatc
     5041 agcgaagcga tgatttttga tctattaaca gatatataaa tgcaaaaact gcataaccac
     5101 tttaactaat actttcaaca ttttcggttt gtattacttc ttattcaaat gtaataaaag
     5161 tatcaacaaa aaattgttaa tatacctcta tactttaacg tcaaggagaa aaaaccccgg
     5221 atccgtactt ccaatccaat attggaagtg gataactctc gaggatcgta atgactagat
     5281 tatataatca gaaatagcct ttggtgatca cttccatttt agtcagcata atagaaagtt
     5341 taattacgcg cttttttaca ttccgacaaa gatctgcccg agaaaattct tcgagaagac
     5401 aacaacaaca aaaaaaataa agcctgggcg aaaaacatcg tgcgtttaca taatatcttg
     5461 aaaagcaaac agatatataa accgctattg aaattacctt tttaagtatt cgcctagaag
     5521 ccttgtttca agtagcatca acaacaggtg ttaggttaaa ctgctttgac tttttgttct
     5581 ttaacgaata tcattaaaaa aaaaggacat ctataaagaa tgtctccaaa ctcgtcgaaa
     5641 actcgaacgg accagataag ctccatgcct ggtattaatg aagccacgaa ggtggaaagt
     5701 aagaatgttg tgaaggatgc tgttccaata aaatctgaag ttgaaaccaa tggtacatca
     5761 atagtaagag aaaaacaaga tccacaggac gggtgtggtc gccatgatcg cgtagtcgat
     5821 agtggctcca agtagcgaag cgagcaggac tgggcggcgg ccaaagcggt cggacagtgc
     5881 tccgagaacg ggtgcgcata gaaattgcat caacgcatat agcgctagca gcacgccata
     5941 gtgactggcg atgctgtcgg aatggacgat atcccgcaag aggccccggc agtaccggca
     6001 taaccaagcc tatgcctaca gccttccagg tgacggtgcc gagattgacc atgaacgcct
     6061 tgttagattt ctacaccggt gcctgactgc gttagcaatt taactgtgat aaactaccgc
     6121 attaaagctt tttctttcca attttttttt tttcgtcatt ataaaaatca ttacgaccga
     6181 gattcccggg taataactga tataattaaa ttgaagctct aatttgtgag tttagtatac
     6241 atgcatttac ttataataca gttttttagt tttgctggcc gcatcttctc aaatatgctt
     6301 cccagcctgc ttttctgtaa cgttcaccct ctaccttagc atcccttccc tttgcaaata
     6361 gtcctcttcc aacaataata atgtcagatc ctgtagagac cacatcatcc acggttctat
     6421 actgttgacc caatgcgtct cccttgtcat ctaaacccac accgggtgtc ataatcaacc
     6481 aatcgtaacc ttcatctctt ccacccatgt ctctttgagc aataaagccg ataacaaaat
     6541 ctttgtcgct cttcgcaatg tcaacagtac ccttagtata ttctccagta gatagggagc
     6601 ccttgcatga caattctgct aacatcaaaa ggcctctagg ttcctttgtt acttcttctg
     6661 ccgcctgctt caaaccgcta acaatacctg ggcccaccac accgtgtgca ttcgtaatgt
     6721 ctgcccattc tgctattctg tatacacccg cagagtactg caatttgact gtattaccaa
     6781 tgtcagcaaa ttttctgtct tcgaagagta aaaaattgta cttggcggat aatgccttta
     6841 gcggcttaac tgtgccctcc atggaaaaat cagtcaagat atccacatgt gtttttagta
     6901 aacaaatttt gggacctaat gcttcaacta actccagtaa ttccttggtg gtacgaacat
     6961 ccaatgaagc acacaagttt gtttgctttt cgtgcatgat attaaatagc ttggcagcaa
     7021 caggactagg atgagtagca gcacgttcct tatatgtagc tttcgacatg atttatcttc
     7081 gtttcctgca ggtttttgtt ctgtgcagtt gggttaagaa tactgggcaa tttcatgttt
     7141 cttcaacact acatatgcgt atatatacca atctaagtct gtgctccttc cttcgttctt
     7201 ccttctgttc ggagattacc gaatcaaaaa aatttcaagg aaaccgaaat caaaaaaaag
     7261 aataaaaaaa aaatgatgaa ttgaaaagct tatcgatgat aagctgtcaa acatgagaat
     7321 tccacggact atagactata ctagtatact ccgtctactg tacgatacac ttccgctcag
     7381 gtccttgtcc tttaacgagg ccttaccact cttttgttac tctattgatc cagctcagca
     7441 aaggcagtgt gatctaagat tctatcttcg cgatgtagta aaactagcta gaccgagaaa
     7501 gagactagaa atgcaaaagg cacttctaca atggctgcca tcattattat ccgatgtgac
     7561 gctgcagctt ctcaatgata ttcgaatacg ctttgaggag atacagccta atatccgaca
     7621 aactgtttta cagatttacg atcgtacttg ttacccatca ttgaattttg aacatccgaa
     7681 cctgggagtt ttccctgaaa cagatagtat atttgaacct gtataataat atatagtcta
     7741 gcgctttacg gaagacaatg tatgtatttc ggttcctgga gaaactattg catctattgc
     7801 ataggtaatc ttgcacgtcg catccccggt tcattttctg cgtttccatc ttgcacttca
     7861 atagcatatc tttgttaacg aagcatctgt gcttcatttt gtagaacaaa aatgcaacgc
     7921 gagagcgcta atttttcaaa caaagaatct gagctgcatt tttacagaac agaaatgcaa
     7981 cgcgaaagcg ctattttacc aacgaagaat ctgtgcttca tttttgtaaa acaaaaatgc
     8041 aacgcgagag cgctaatttt tcaaacaaag aatctgagct gcatttttac agaacagaaa
     8101 tgcaacgcga gagcgctatt ttaccaacaa agaatctata cttctttttt gttctacaaa
     8161 aatgcatccc gagagcgcta tttttctaac aaagcatctt agattacttt ttttctcctt
     8221 tgtgcgctct ataatgcagt ctcttgataa ctttttgcac tgtaggtccg ttaaggttag
     8281 aagaaggcta ctttggtgtc tattttctct tccataaaaa aagcctgact ccacttcccg
     8341 cgtttactga ttactagcga agctgcgggt gcattttttc aagataaagg catccccgat
     8401 tatattctat accgtcacaa agtatgcgca atccacatac agaaagtgat agcgttgatg
     8461 attcttcatt ggtcagaaaa ttatgaacgg tttcttctat tttgtctcta tatactacgt
     8521 ataggaaatg tttacatttt cgtattgttt tcgattcact ctatgaatag ttcttactac
     8581 aatttttttg tctaaagagt aatactagag ataaacataa aaaatgtaga ggtcgagttt
     8641 agatgcaagt tcaaggagcg aaaggtggat gggtaggtta tatagggata tagcacagag
     8701 atatatagca aagagatact tttgagcaat gtttgtggaa gcggtattcg cattatttta
     8761 gtagctcgtt acagtccggt gcgtttttgg ttttttgaaa gtgcgtcttc agagcgcttt
     8821 tggttttcaa aagcgctctg aagttcctat actttctaga gaataggaac ttc
//
