LOCUS       sequence-108417-        5993 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses destabilized Citrine in yeast..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5993)
  AUTHORS   Gnugge R, Liphardt T, Rudolf F
  TITLE     A shuttle vector series for precise genetic engineering of 
            Saccharomyces cerevisiae.
  JOURNAL   Yeast. 2015 Dec 9. doi: 10.1002/yea.3144.
  PUBMED    26647923
REFERENCE   2  (bases 1 to 5993)
  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
            Dec 9. doi: 10.1002/yea.3144."
COMMENT     Sequence Label: FRP1432
FEATURES             Location/Qualifiers
     source          1..5993
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(374..1177)
                     /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
                     ELVEALGPKICLLKTHVDLLTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSACVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLATGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     promoter        complement(1178..1394)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     1445..1467
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1459..1476
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1460..1476
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1483..1502
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1483..1501
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        1621..2287
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase; also known as the TDH3 promoter"
     primer_bind     2157..2180
                     /label=GPDpro-F
                     /note="S. cerevisiae GPD promoter, forward primer"
     CDS             2330..3046
                     /codon_start=1
                     /product="yeast-enhanced green fluorescent protein"
                     /label=yeGFP
                     /note="codons optimized for translation in S. cerevisiae 
                     and C. albicans"
                     /translation="MRSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     terminator      3264..3513
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(3280..3298)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     promoter        complement(3676..3694)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3715..3731)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3715..3731)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3728..3750)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3739..3755
                     /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     complement(4065..4082)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4236..4824)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4316..4335)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5003..5794,5795..5863))
                     /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     5626..5645
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5864..5968)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacgtcgaaa gaacgcggca ttagcaacat ttgcgaagcg aacgattttg cgaactgcta
       61 ttaattaact ctggcgcgcc actacttttt ggaactcttg ttgttctttg gagttcaatg
      121 cgtccatctt tacagtcctg tcttattgtt cttgatttgt gccccgtaaa atactgttac
      181 ttggttctgg cgaggtattg gatagttcct ttttataaag gccatgaagg tttttctttc
      241 caattttttt tttttcgtca ttatagaaat cattacgacc gagattcccg gcatatggta
      301 ataactgata taattaaatt gaagctctaa tttgtgagtt tagtatacat gcatttactt
      361 ataatacagt tttttagttt tgctggccgc atcttctcaa atatgcttcc cagcctgctt
      421 ttctgtaacg ttcaccctct accttagcat cccttccctt tgcaaatagt cctcttccaa
      481 caataataat gtcagatcct gtagagacca catcatccac ggttctatac tgttgaccca
      541 atgcgtctcc cttgtcatct aaacccacac cgggtgtcat aatcaaccaa tcgtaacctt
      601 catctcttcc acccatgtct ctttgagcaa taaagccgat aacaaaatct ttgtcgctct
      661 tcgcaatgtc aacagtaccc ttagtatatt ctccagtagc tagggagccc ttgcatgaca
      721 attctgctaa catcaaaagg cctctaggtt cctttgttac ttcttccgcc gcctgcttca
      781 aaccgctaac aatacctggg cccaccacac cgtgtgcatt cgtaatgtct gcccattctg
      841 ctattctgta tacacacgca gagtactgca atttgactgt attaccaatg tcagcaaatt
      901 ttctgtcttc gaagagtaaa aaattgtact tggcggataa tgcctttagc ggcttaactg
      961 tgccctccat ggaaaaatca gtcaagagat ccacatgtgt ttttagtaaa caaattttgg
     1021 gacctaatgc ttcaactaac tccagtaatt ccttggtggt acgaacatcc aatgaagcac
     1081 acaagtttgt ttgcttttcg tgcatgatat taaatagctt ggcagcaaca ggactaggat
     1141 gagtagcagc acgttcctta tatgtagctt tcgacatgat ttatcttcgt ttcctgcacg
     1201 tttttgttct gtgcagttgg gttaagaata ctgggcaatt tcatgtttct tcaacaccac
     1261 ctatgcgtat atataccaat ctaagtctgt gctccttcct tcgttcttcc ttctgctcgg
     1321 agattaccga atcaaaaaaa tttcaaagaa accggaatca aaaaaaagaa caaaaaaaaa
     1381 aaagatgaat gaaaagcgat cggatgtgct gcaaggcgat taagttgggt aacgccaggg
     1441 ttttcccagt cacgacgttg taaaacgacg gccagtgaat tgtaatacga ctcactatag
     1501 ggcgaattgg agaaacagaa taatatataa acctgtataa tataaccttg aagactatat
     1561 ttcttttcgc ggccgcctat agggcgaatt ggagctcccg ctctagacag ttcgagttta
     1621 tcattatcaa tactgccatt tcaaagaata cgtaaataat taatagtagt gattttccta
     1681 actttattta gtcaaaaaat tagcctttta attctgctgt aacccgtaca tgcccaaaat
     1741 agggggcggg ttacacagaa tatataacat cgtaggtgtc tgggtgaaca gtttattcct
     1801 ggcatccact aaatataatg gagcccgctt tttaagctgg catccagaaa aaaaaagaat
     1861 cccagcacca aaatattgtt ttcttcacca accatcagtt cataggtcca ttctcttagc
     1921 gcaactacag agaacagggg cacaaacagg caaaaaacgg gcacaacctc aatggagtga
     1981 tgcaacctgc ctggagtaaa tgatgacaca aggcaattga cccacgcatg tatctatctc
     2041 attttcttac accttctatt accttctgct ctctctgatt tggaaaaagc tgaaaaaaaa
     2101 ggttgaaacc agttccctga aattattccc ctacttgact aataagtata taaagacggt
     2161 aggtattgat tgtaattctg taaatctatt tcttaaactt cttaaattct acttttatag
     2221 ttagtctttt ttttagtttt aaaacaccaa gaacttagtt tcgaataaac acacataaac
     2281 aaacaaaact agtggatccc ccgggctgca ggaattcgat atcaagctta tgagatctaa
     2341 aggtgaagaa ttattcactg gtgttgtccc aattttggtt gaattagatg gtgatgttaa
     2401 tggtcacaaa ttttctgtct ccggtgaagg tgaaggtgat gctacttacg gtaaattgac
     2461 cttaaaattt atttgtacta ctggtaaatt gccagttcca tggccaacct tagtcactac
     2521 tttaggttat ggtttgatgt gttttgctag atacccagat catatgaaac aacatgactt
     2581 tttcaagtct gccatgccag aaggttatgt tcaagaaaga actatttttt tcaaagatga
     2641 cggtaactac aagaccagag ctgaagtcaa gtttgaaggt gataccttag ttaatagaat
     2701 cgaattaaaa ggtattgatt ttaaagaaga tggtaacatt ttaggtcaca aattggaata
     2761 caactataac tctcacaatg tttacatcat ggctgacaaa caaaagaatg gtatcaaagt
     2821 taacttcaaa attagacaca acattgaaga tggttctgtt caattagctg accattatca
     2881 acaaaatact ccaattggtg atggtccagt cttgttacca gacaaccatt acttatccta
     2941 tcaatctaag ttatccaaag atccaaacga aaagagagac cacatggtct tgttagaatt
     3001 tgttactgct gctggtatta cccatggtat ggatgaattg tacaaaggat ccacttggtc
     3061 tactgttaga agatccggtt tggaaactca attcagagct ttggacttct tcgccagagg
     3121 tttggtcaag tctccaatca aggttgtcgg cttgtctacc ttgccagaaa tttacgaaaa
     3181 gatggaaaag ggtcaaatcg ttggtagata cgttgttgac acttctaaat aactcgagac
     3241 aggccccttt tcctttgtct atatcatgta attagttatg tcacgcttac attcacgccc
     3301 tcctcccaca tccgctctaa ccgaaaagga aggagttaga caacctgaag tctaggtccc
     3361 tatttatttt ttttaatagt tatgttagta ttaagaacgt tatttatatt tcaaattttt
     3421 cttttttttc tgtacaaacg cgtgtacgca tgtaacatta tactgaaaac cttgcttgag
     3481 aaggttttgg gacgctcgaa ggctttaatt tgcaaggttc gcagtttaca ctctcatcgt
     3541 cgctctcatc atcgcttccg ttgttgtttt ccttagtagc gtctgcttgg tacccagctt
     3601 cggccgtatt tacagataac gatgtcgtta ttatatatat atatatatat tgtcacttgt
     3661 acccagcttt tgttcccttt agtgagggtt aatttcgagc ttggcgtaat catggtcata
     3721 gctgtttcct gtgtgaaatt gttatccgct cacaacagct ggatgaattg aatagcttta
     3781 tggaccctga aaccacagcc acattaacct tctttgatgg tcaaaactta tccttcacca
     3841 taaatatgcc tcgcaaaaaa ggtaattaac atatatagaa ttacattatt tatgaaatat
     3901 catcactatc tcttagcatc tttaatcctt ttctacatca gataacttcg gtttgttatc
     3961 atcgtctgta ttgtcatcaa ttggcgcgcc tctccctgca ggcattaatg aatcggccaa
     4021 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     4081 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     4141 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     4201 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     4261 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     4321 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     4381 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     4441 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     4501 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     4561 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     4621 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     4681 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     4741 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     4801 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttgtttaaa cctacggggt
     4861 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     4921 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     4981 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     5041 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     5101 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     5161 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     5221 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     5281 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     5341 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     5401 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatggtt gtcagaagta
     5461 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     5521 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     5581 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     5641 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     5701 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     5761 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     5821 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     5881 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     5941 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cct
//
