LOCUS       sequence-113667-        7351 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 7351)
  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 7351)
  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-G1
FEATURES             Location/Qualifiers
     source          1..7351
                     /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        196..416
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             417..1220
                     /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(1351..1806)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1488..1509)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1700..1719
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     terminator      complement(1885..2050)
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     1971..1995
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     CDS             complement(2198..2221)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        complement(2255..2661)
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     primer_bind     3663..3682
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3663..3681
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             3699..3728
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     terminator      3758..3947
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(4019..4036)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4190..4778)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4270..4289)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4949..5740,5741..5809))
                     /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     5572..5591
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5810..5914)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5941..7283
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(6904..6951)
                     /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     7324..7342
                     /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 agcttttcaa ttcaattcat catttttttt ttattctttt ttttgatttc
      241 ggtttctttg aaattttttt gattcggtaa tctccgaaca gaaggaagaa cgaaggaagg
      301 agcacagact tagattggta tatatacgca tatgtagtgt tgaagaaaca tgaaattgcc
      361 cagtattctt aacccaactg cacagaacaa aaacctgcag gaaacgaaga taaatcatgt
      421 cgaaagctac atataaggaa cgtgctgcta ctcatcctag tcctgttgct gccaagctat
      481 ttaatatcat gcacgaaaag caaacaaact tgtgtgcttc attggatgtt cgtaccacca
      541 aggaattact ggagttagtt gaagcattag gtcccaaaat ttgtttacta aaaacacatg
      601 tggatatctt gactgatttt tccatggagg gcacagttaa gccgctaaag gcattatccg
      661 ccaagtacaa ttttttactc ttcgaagaca gaaaatttgc tgacattggt aatacagtca
      721 aattgcagta ctctgcgggt gtatacagaa tagcagaatg ggcagacatt acgaatgcac
      781 acggtgtggt gggcccaggt attgttagcg gtttgaagca ggcggcagaa gaagtaacaa
      841 aggaacctag aggccttttg atgttagcag aattgtcatg caagggctcc ctatctactg
      901 gagaatatac taagggtact gttgacattg cgaagagcga caaagatttt gttatcggct
      961 ttattgctca aagagacatg ggtggaagag atgaaggtta cgattggttg attatgacac
     1021 ccggtgtggg tttagatgac aagggagacg cattgggtca acagtataga accgtggatg
     1081 atgtggtctc tacaggatct gacattatta ttgttggaag aggactattt gcaaagggaa
     1141 gggatgctaa ggtagagggt gaacgttaca gaaaagcagg ctgggaagca tatttgagaa
     1201 gatgcggcca gcaaaactaa aaaactgtat tataagtaaa tgcatgtata ctaaactcac
     1261 aaattagagc ttcaatttaa ttatatcagt tattacccta tgcggtgtga aataccgcac
     1321 agatgcgtaa ggagaaaata ccgcatcagg aaattgtaaa cgttaatatt ttgttaaaat
     1381 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa
     1441 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca
     1501 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
     1561 gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta
     1621 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg
     1681 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa
     1741 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
     1801 gcgcgtccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc
     1861 ttcgctatta cgccagctga attggagcga cctcatgcta tacctgagaa agcaacctga
     1921 cctacaggaa agagttactc aagaataaga attttcgttt taaaacctaa gagtcacttt
     1981 aaaatttgta tacacttatt ttttttataa cttatttaat aataaaaatc ataaatcata
     2041 agaaattcgc ttatttagaa gtgtcaacaa cgtatctacc aacgatttga cccttttcca
     2101 tcttttcgta aatttctggc aaggtagaca agccgacaac cttgattgga gacttgacca
     2161 aacctctggc gaagaattgt taattaagag ctcagatctt atcgtcgtca tccttgtaat
     2221 ccatcgatac tagtgcggcc gcttgtaatt aaaacttaga ttagattgct atgctttctt
     2281 tctaatgagc aagaagtaaa aaaagttgta atagaacaag aaaaatgaaa ctgaaacttg
     2341 agaaattgaa gaccgtttat taacttaaat atcaatggga ggtcatcgaa agagaaaaaa
     2401 atcaaaaaaa aaaattttca agaaaaagaa acgtgataaa aatttttatt gcctttttcg
     2461 acgaagaaaa agaaacgagg cggtctcttt tttcttttcc aaacctttag tacgggtaat
     2521 taacgacacc ctagaggaag aaagagggga aatttagtat gctgtgcttg ggtgttttga
     2581 agtggtacgg cgatgcgcgg agtccgagaa aatctggaag agtaaaaaag gagtagaaac
     2641 attttgaagc tatggtgtgt gcggccggcc tggaagtacc ttcaaagaat ggggtcttat
     2701 cttgttttgc aagtaccact gagcaggata ataatagaaa tgataatata ctatagtaga
     2761 gataacgtcg atgacttccc atactgtaat tgcttttagt tgtgtatttt tagtgtgcaa
     2821 gtttctgtaa atcgattaat ttttttttct ttcctctttt tattaacctt aatttttatt
     2881 ttagattcct gacttcaact caagacgcac agatattata acatctgcat aataggcatt
     2941 tgcaagaatt actcgtgagt aaggaaagag tgaggaacta tcgcatacct gcatttaaag
     3001 atgccgattt gggcgcgaat cctttatttt ggcttcaccc tcatactatt atcagggcca
     3061 gaaaaaggaa gtgtttccct ccttcttgaa ttgatgttac cctcataaag cacgtggcct
     3121 cttatcgaga aagaaattac cgtcgctcgt gatttgtttg caaaaagaac aaaactgaaa
     3181 aaacccagac acgctcgact tcctgtcttc ctattgattg cagcttccaa tttcgtcaca
     3241 caacaaggtc ctagcgacgg ctcacaggtt ttgtaacaag caatcgaagg ttctggaatg
     3301 gcgggaaagg gtttagtacc acatgctatg atgcccactg tgatctccag agcaaagttc
     3361 gttcgatcgt actgttactc tctctctttc aaacagaatt gtccgaatcg tgtgacaaca
     3421 acagcctgtt ctcacacact cttttcttct aaccaagggg gtggtttagt ttagtagaac
     3481 ctcgtgaaac ttacatttac atatatataa acttgcataa attggtcaat gcaagaaata
     3541 catatttggt cttttctaat tcgtagtttt tcaagttctt agatgctttc tttttctctt
     3601 ttttacagat catcaaggaa gtaattatct actttttaca acaaatataa aacaaggatc
     3661 cgtaatacga ctcactatag ggcccgggcg tcgacatgga acagaagttg atttccgaag
     3721 aagacctcga gtaagcttgg taccgcggct agctaagatc cgctctaacc gaaaaggaag
     3781 gagttagaca acctgaagtc taggtcccta tttatttttt tatagttatg ttagtattaa
     3841 gaacgttatt tatatttcaa atttttcttt tttttctgta cagacgcgtg tacgcatgta
     3901 acattatact gaaaaccttg cttgagaagg ttttgggacg ctcgaagatc cagctgcatt
     3961 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     4021 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     4081 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     4141 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     4201 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     4261 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     4321 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     4381 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     4441 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     4501 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     4561 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     4621 acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     4681 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     4741 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     4801 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     4861 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     4921 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     4981 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     5041 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     5101 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     5161 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     5221 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     5281 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     5341 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     5401 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     5461 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     5521 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     5581 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     5641 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     5701 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     5761 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     5821 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     5881 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     5941 aacgaagcat ctgtgcttca ttttgtagaa caaaaatgca acgcgagagc gctaattttt
     6001 caaacaaaga atctgagctg catttttaca gaacagaaat gcaacgcgaa agcgctattt
     6061 taccaacgaa gaatctgtgc ttcatttttg taaaacaaaa atgcaacgcg agagcgctaa
     6121 tttttcaaac aaagaatctg agctgcattt ttacagaaca gaaatgcaac gcgagagcgc
     6181 tattttacca acaaagaatc tatacttctt ttttgttcta caaaaatgca tcccgagagc
     6241 gctatttttc taacaaagca tcttagatta ctttttttct cctttgtgcg ctctataatg
     6301 cagtctcttg ataacttttt gcactgtagg tccgttaagg ttagaagaag gctactttgg
     6361 tgtctatttt ctcttccata aaaaaagcct gactccactt cccgcgttta ctgattacta
     6421 gcgaagctgc gggtgcattt tttcaagata aaggcatccc cgattatatt ctataccgat
     6481 gtggattgcg catactttgt gaacagaaag tgatagcgtt gatgattctt cattggtcag
     6541 aaaattatga acggtttctt ctattttgtc tctatatact acgtatagga aatgtttaca
     6601 ttttcgtatt gttttcgatt cactctatga atagttctta ctacaatttt tttgtctaaa
     6661 gagtaatact agagataaac ataaaaaatg tagaggtcga gtttagatgc aagttcaagg
     6721 agcgaaaggt ggatgggtag gttatatagg gatatagcac agagatatat agcaaagaga
     6781 tacttttgag caatgtttgt ggaagcggta ttcgcaatat tttagtagct cgttacagtc
     6841 cggtgcgttt ttggtttttt gaaagtgcgt cttcagagcg cttttggttt tcaaaagcgc
     6901 tctgaagttc ctatactttc tagagaatag gaacttcgga ataggaactt caaagcgttt
     6961 ccgaaaacga gcgcttccga aaatgcaacg cgagctgcgc acatacagct cactgttcac
     7021 gtcgcaccta tatctgcgtg ttgcctgtat atatatatac atgagaagaa cggcatagtg
     7081 cgtgtttatg cttaaatgcg tacttatatg cgtctattta tgtaggatga aaggtagtct
     7141 agtacctcct gtgatattat cccattccat gcggggtatc gtatgcttcc ttcagcacta
     7201 ccctttagct gttctatatg ctgccactcc tcaattggat tagtctcatc cttcaatgct
     7261 atcatttcct ttgatattgg atcatactaa gaaaccatta ttatcatgac attaacctat
     7321 aaaaataggc gtatcacgag gccctttcgt c
//
