LOCUS       sequence-459084-        7344 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast expression plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7344)
  AUTHORS   Partow S, Siewers V, Bjorn S, Nielsen J, Maury J
  TITLE     Characterization of different promoters for designing a new 
            expression vector in Saccharomyces cerevisiae.
  JOURNAL   Yeast. 2010 Nov;27(11):955-64. doi: 10.1002/yea.1806.
  PUBMED    20625983
REFERENCE   2  (bases 1 to 7344)
  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"; doi: "10.1002/yea"; 
            journalName: "Yeast"; date: "2010-11"; volume: "27"; issue: "11"; 
            pages: "955-64"
COMMENT     Sequence Label: pSP-G2
FEATURES             Location/Qualifiers
     source          1..7344
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        194..409
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             410..1213
                     /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"
     rep_origin      complement(1344..1799)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1481..1502)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1693..1712
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     terminator      complement(1878..2043)
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     1964..1988
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     CDS             complement(2191..2214)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        3230..3636
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     primer_bind     3656..3675
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3656..3674
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             3692..3721
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     terminator      3751..3940
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(4012..4029)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4183..4771)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4263..4282)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4942..5733,5734..5802))
                     /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     5565..5584
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5803..5907)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5934..7276
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(6897..6944)
                     /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)."
     primer_bind     7317..7335
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accataccac cttttcaatt catcattttt tttttattct tttttttgat ttcggtttcc
      241 ttgaaatttt tttgattcgg taatctccga acagaaggaa gaacgaagga aggagcacag
      301 acttagattg gtatatatac gcatatgtag tgttgaagaa acatgaaatt gcccagtatt
      361 cttaacccaa ctgcacagaa caaaaacctg caggaaacga agataaatca tgtcgaaagc
      421 tacatataag gaacgtgctg ctactcatcc tagtcctgtt gctgccaagc tatttaatat
      481 catgcacgaa aagcaaacaa acttgtgtgc ttcattggat gttcgtacca ccaaggaatt
      541 actggagtta gttgaagcat taggtcccaa aatttgttta ctaaaaacac atgtggatat
      601 cttgactgat ttttccatgg agggcacagt taagccgcta aaggcattat ccgccaagta
      661 caatttttta ctcttcgaag acagaaaatt tgctgacatt ggtaatacag tcaaattgca
      721 gtactctgcg ggtgtataca gaatagcaga atgggcagac attacgaatg cacacggtgt
      781 ggtgggccca ggtattgtta gcggtttgaa gcaggcggca gaagaagtaa caaaggaacc
      841 tagaggcctt ttgatgttag cagaattgtc atgcaagggc tccctatcta ctggagaata
      901 tactaagggt actgttgaca ttgcgaagag cgacaaagat tttgttatcg gctttattgc
      961 tcaaagagac atgggtggaa gagatgaagg ttacgattgg ttgattatga cacccggtgt
     1021 gggtttagat gacaagggag acgcattggg tcaacagtat agaaccgtgg atgatgtggt
     1081 ctctacagga tctgacatta ttattgttgg aagaggacta tttgcaaagg gaagggatgc
     1141 taaggtagag ggtgaacgtt acagaaaagc aggctgggaa gcatatttga gaagatgcgg
     1201 ccagcaaaac taaaaaactg tattataagt aaatgcatgt atactaaact cacaaattag
     1261 agcttcaatt taattatatc agttattacc ctatgcggtg tgaaataccg cacagatgcg
     1321 taaggagaaa ataccgcatc aggaaattgt aagcgttaat attttgttaa aattcgcgtt
     1381 aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta
     1441 taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc
     1501 actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg
     1561 cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact
     1621 aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt
     1681 ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc
     1741 ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc
     1801 cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta
     1861 ttacgccagc tgaattggag cgacctcatg ctatacctga gaaagcaacc tgacctacag
     1921 gaaagagtta ctcaagaata agaattttcg ttttaaaacc taagagtcac tttaaaattt
     1981 gtatacactt atttttttta taacttattt aataataaaa atcataaatc ataagaaatt
     2041 cgcttattta gaagtgtcaa caacgtatct accaacgatt tgaccctttt ccatcttttc
     2101 gtaaatttct ggcaaggtag acaagccgac aaccttgatt ggagacttga ccaaacctct
     2161 ggcgaagaat tgttaattaa gagctcagat cttatcgtcg tcatccttgt aatccatcga
     2221 tactagtgcg gccgcttgtt ttatatttgt tgtaaaaagt agataattac ttccttgatg
     2281 atctgtaaaa aagagaaaaa gaaagcatct aagaacttga aaaactacga attagaaaag
     2341 accaaatatg tatttcttgc attgaccaat ttatgcaagt ttatatatat gtaaatgtaa
     2401 gtttcacgag gttctactaa actaaaccac ccccttggtt agaagaaaag agtgtgtgag
     2461 aacaggctgt tgttgtcaca cgattcggac aattctgttt gaaagagaga gagtaacagt
     2521 acgatcgaac gaactttgct ctggagatca cagtgggcat catagcatgt ggtactaaac
     2581 cctttcccgc cattccagaa ccttcgattg cttgttacaa aacctgtgag ccgtcgctag
     2641 gaccttgttg tgtgacgaaa ttggaagctg caatcaatag gaagacagga agtcgagcgt
     2701 gtctgggttt tttcagtttt gttctttttg caaacaaatc acgagcgacg gtaatttctt
     2761 tctcgataag aggccacgtg ctttatgagg gtaacatcaa ttcaagaagg agggaaacac
     2821 ttcctttttc tggccctgat aatagtatga gggtgaagcc aaaataaagg attcgcgccc
     2881 aaatcggcat ctttaaatgc aggtatgcga tagttcctca ctctttcctt actcacgagt
     2941 aattcttgca aatgcctatt atgcagatgt tataatatct gtgcgtcttg agttgaagtc
     3001 aggaatctaa aataaaaatt aaggttaata aaaagaggaa agaaaaaaaa attaatcgat
     3061 ttacagaaac ttgcacacta aaaatacaca actaaaagca attacagtat gggaagtcat
     3121 cgacgttatc tctactatag tatattatca tttctattat tatcctgctc agtggtactt
     3181 gcaaaacaag ataagacccc attctttgaa ggtacttcca ggccggccgc acacaccata
     3241 gcttcaaaat gtttctactc cttttttact cttccagatt ttctcggact ccgcgcatcg
     3301 ccgtaccact tcaaaacacc caagcacagc atactaaatt tcccctcttt cttcctctag
     3361 ggtgtcgtta attacccgta ctaaaggttt ggaaaagaaa aaagagaccg cctcgtttct
     3421 ttttcttcgt cgaaaaaggc aataaaaatt tttatcacgt ttctttttct tgaaaatttt
     3481 tttttttgat ttttttctct ttcgatgacc tcccattgat atttaagtta ataaacggtc
     3541 ttcaatttct caagtttcag tttcattttt cttgttctat tacaactttt tttacttctt
     3601 gctcattaga aagaaagcat agcaatctaa tctaagtttt aattacaagg atccgtaata
     3661 cgactcacta tagggcccgg gcgtcgacat ggaacagaag ttgatttccg aagaagacct
     3721 cgagtaagct tggtaccgcg gctagctaag atccgctcta accgaaaagg aaggagttag
     3781 acaacctgaa gtctaggtcc ctatttattt ttttatagtt atgttagtat taagaacgtt
     3841 atttatattt caaatttttc ttttttttct gtacagacgc gtgtacgcat gtaacattat
     3901 actgaaaacc ttgcttgaga aggttttggg acgctcgaag atccagctgc attaatgaat
     3961 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     4021 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     4081 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     4141 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     4201 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     4261 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     4321 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     4381 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     4441 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     4501 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     4561 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     4621 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     4681 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     4741 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     4801 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     4861 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     4921 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     4981 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     5041 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     5101 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     5161 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     5221 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     5281 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     5341 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     5401 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     5461 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     5521 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     5581 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     5641 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     5701 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     5761 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     5821 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     5881 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgaacgaag
     5941 catctgtgct tcattttgta gaacaaaaat gcaacgcgag agcgctaatt tttcaaacaa
     6001 agaatctgag ctgcattttt acagaacaga aatgcaacgc gaaagcgcta ttttaccaac
     6061 gaagaatctg tgcttcattt ttgtaaaaca aaaatgcaac gcgagagcgc taatttttca
     6121 aacaaagaat ctgagctgca tttttacaga acagaaatgc aacgcgagag cgctatttta
     6181 ccaacaaaga atctatactt cttttttgtt ctacaaaaat gcatcccgag agcgctattt
     6241 ttctaacaaa gcatcttaga ttactttttt tctcctttgt gcgctctata atgcagtctc
     6301 ttgataactt tttgcactgt aggtccgtta aggttagaag aaggctactt tggtgtctat
     6361 tttctcttcc ataaaaaaag cctgactcca cttcccgcgt ttactgatta ctagcgaagc
     6421 tgcgggtgca ttttttcaag ataaaggcat ccccgattat attctatacc gatgtggatt
     6481 gcgcatactt tgtgaacaga aagtgatagc gttgatgatt cttcattggt cagaaaatta
     6541 tgaacggttt cttctatttt gtctctatat actacgtata ggaaatgttt acattttcgt
     6601 attgttttcg attcactcta tgaatagttc ttactacaat ttttttgtct aaagagtaat
     6661 actagagata aacataaaaa atgtagaggt cgagtttaga tgcaagttca aggagcgaaa
     6721 ggtggatggg taggttatat agggatatag cacagagata tatagcaaag agatactttt
     6781 gagcaatgtt tgtggaagcg gtattcgcaa tattttagta gctcgttaca gtccggtgcg
     6841 tttttggttt tttgaaagtg cgtcttcaga gcgcttttgg ttttcaaaag cgctctgaag
     6901 ttcctatact ttctagagaa taggaacttc ggaataggaa cttcaaagcg tttccgaaaa
     6961 cgagcgcttc cgaaaatgca acgcgagctg cgcacataca gctcactgtt cacgtcgcac
     7021 ctatatctgc gtgttgcctg tatatatata tacatgagaa gaacggcata gtgcgtgttt
     7081 atgcttaaat gcgtacttat atgcgtctat ttatgtagga tgaaaggtag tctagtacct
     7141 cctgtgatat tatcccattc catgcggggt atcgtatgct tccttcagca ctacccttta
     7201 gctgttctat atgctgccac tcctcaattg gattagtctc atccttcaat gctatcattt
     7261 cctttgatat tggatcatac taagaaacca ttattatcat gacattaacc tataaaaata
     7321 ggcgtatcac gaggcccttt cgtc
//
