LOCUS       sequence-108422-        6458 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 6458)
  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 6458)
  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: FRP1454
FEATURES             Location/Qualifiers
     source          1..6458
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    18..521
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     CDS             complement(839..1642)
                     /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(1643..1859)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     1910..1932
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1924..1941
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1925..1941
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1948..1967
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1948..1966
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        2086..2752
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase; also known as the TDH3 promoter"
     primer_bind     2622..2645
                     /label=GPDpro-F
                     /note="S. cerevisiae GPD promoter, forward primer"
     CDS             2795..3511
                     /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      3729..3978
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(3745..3763)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     promoter        complement(4141..4159)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4180..4196)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4180..4196)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4193..4215)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4204..4220
                     /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(4530..4547)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4701..5289)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4781..4800)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5468..6259,6260..6328))
                     /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     6091..6110
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6329..6433)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacgtcgggt ccttttcatc acgtgctata aaaataatta taatttaaat tttttaatat
       61 aaatatataa attaaaaata gaaagtaaaa aaagaaatta aagaaaaaat agtttttgtt
      121 ttccgaagat gtaaaagact ctagggggat cgccaacaaa tactaccttt tatcttgctc
      181 ttcctgctct caggtattaa tgccgaattg tttcatcttg tctgtgtaga agaccacaca
      241 cgaaaatcct gtgattttac attttactta tcgttaatcg aatgtatatc tatttaatct
      301 gcttttcttg tctaataaat atatatgtaa agtacgcttt ttgttgaaat tttttaaacc
      361 tttgtttatt tttttttctt cattccgtaa ctcttctacc ttctttattt actttctaaa
      421 atccaaatac aaaacataaa aataaataaa cacagagtaa attcccaaat tattccatca
      481 ttaaaagata cgaggcgcgt gtaagttaca ggcaagcgat ccgtcttaat taactctggc
      541 gcgccactac tttttggaac tcttgttgtt ctttggagtt caatgcgtcc atctttacag
      601 tcctgtctta ttgttcttga tttgtgcccc gtaaaatact gttacttggt tctggcgagg
      661 tattggatag ttccttttta taaaggccat gaaggttttt ctttccaatt tttttttttt
      721 cgtcattata gaaatcatta cgaccgagat tcccggcata tggtaataac tgatataatt
      781 aaattgaagc tctaatttgt gagtttagta tacatgcatt tacttataat acagtttttt
      841 agttttgctg gccgcatctt ctcaaatatg cttcccagcc tgcttttctg taacgttcac
      901 cctctacctt agcatccctt ccctttgcaa atagtcctct tccaacaata ataatgtcag
      961 atcctgtaga gaccacatca tccacggttc tatactgttg acccaatgcg tctcccttgt
     1021 catctaaacc cacaccgggt gtcataatca accaatcgta accttcatct cttccaccca
     1081 tgtctctttg agcaataaag ccgataacaa aatctttgtc gctcttcgca atgtcaacag
     1141 tacccttagt atattctcca gtagctaggg agcccttgca tgacaattct gctaacatca
     1201 aaaggcctct aggttccttt gttacttctt ccgccgcctg cttcaaaccg ctaacaatac
     1261 ctgggcccac cacaccgtgt gcattcgtaa tgtctgccca ttctgctatt ctgtatacac
     1321 acgcagagta ctgcaatttg actgtattac caatgtcagc aaattttctg tcttcgaaga
     1381 gtaaaaaatt gtacttggcg gataatgcct ttagcggctt aactgtgccc tccatggaaa
     1441 aatcagtcaa gagatccaca tgtgttttta gtaaacaaat tttgggacct aatgcttcaa
     1501 ctaactccag taattccttg gtggtacgaa catccaatga agcacacaag tttgtttgct
     1561 tttcgtgcat gatattaaat agcttggcag caacaggact aggatgagta gcagcacgtt
     1621 ccttatatgt agctttcgac atgatttatc ttcgtttcct gcacgttttt gttctgtgca
     1681 gttgggttaa gaatactggg caatttcatg tttcttcaac accacctatg cgtatatata
     1741 ccaatctaag tctgtgctcc ttccttcgtt cttccttctg ctcggagatt accgaatcaa
     1801 aaaaatttca aagaaaccgg aatcaaaaaa aagaacaaaa aaaaaaaaga tgaatgaaaa
     1861 gcgatcggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc ccagtcacga
     1921 cgttgtaaaa cgacggccag tgaattgtaa tacgactcac tatagggcga attggagaaa
     1981 cagaataata tataaacctg tataatataa ccttgaagac tatatttctt ttcgcggccg
     2041 cctatagggc gaattggagc tcccgctcta gacagttcga gtttatcatt atcaatactg
     2101 ccatttcaaa gaatacgtaa ataattaata gtagtgattt tcctaacttt atttagtcaa
     2161 aaaattagcc ttttaattct gctgtaaccc gtacatgccc aaaatagggg gcgggttaca
     2221 cagaatatat aacatcgtag gtgtctgggt gaacagttta ttcctggcat ccactaaata
     2281 taatggagcc cgctttttaa gctggcatcc agaaaaaaaa agaatcccag caccaaaata
     2341 ttgttttctt caccaaccat cagttcatag gtccattctc ttagcgcaac tacagagaac
     2401 aggggcacaa acaggcaaaa aacgggcaca acctcaatgg agtgatgcaa cctgcctgga
     2461 gtaaatgatg acacaaggca attgacccac gcatgtatct atctcatttt cttacacctt
     2521 ctattacctt ctgctctctc tgatttggaa aaagctgaaa aaaaaggttg aaaccagttc
     2581 cctgaaatta ttcccctact tgactaataa gtatataaag acggtaggta ttgattgtaa
     2641 ttctgtaaat ctatttctta aacttcttaa attctacttt tatagttagt ctttttttta
     2701 gttttaaaac accaagaact tagtttcgaa taaacacaca taaacaaaca aaactagtgg
     2761 atcccccggg ctgcaggaat tcgatatcaa gcttatgaga tctaaaggtg aagaattatt
     2821 cactggtgtt gtcccaattt tggttgaatt agatggtgat gttaatggtc acaaattttc
     2881 tgtctccggt gaaggtgaag gtgatgctac ttacggtaaa ttgaccttaa aatttatttg
     2941 tactactggt aaattgccag ttccatggcc aaccttagtc actactttag gttatggttt
     3001 gatgtgtttt gctagatacc cagatcatat gaaacaacat gactttttca agtctgccat
     3061 gccagaaggt tatgttcaag aaagaactat ttttttcaaa gatgacggta actacaagac
     3121 cagagctgaa gtcaagtttg aaggtgatac cttagttaat agaatcgaat taaaaggtat
     3181 tgattttaaa gaagatggta acattttagg tcacaaattg gaatacaact ataactctca
     3241 caatgtttac atcatggctg acaaacaaaa gaatggtatc aaagttaact tcaaaattag
     3301 acacaacatt gaagatggtt ctgttcaatt agctgaccat tatcaacaaa atactccaat
     3361 tggtgatggt ccagtcttgt taccagacaa ccattactta tcctatcaat ctaagttatc
     3421 caaagatcca aacgaaaaga gagaccacat ggtcttgtta gaatttgtta ctgctgctgg
     3481 tattacccat ggtatggatg aattgtacaa aggatccact tggtctactg ttagaagatc
     3541 cggtttggaa actcaattca gagctttgga cttcttcgcc agaggtttgg tcaagtctcc
     3601 aatcaaggtt gtcggcttgt ctaccttgcc agaaatttac gaaaagatgg aaaagggtca
     3661 aatcgttggt agatacgttg ttgacacttc taaataactc gagacaggcc ccttttcctt
     3721 tgtctatatc atgtaattag ttatgtcacg cttacattca cgccctcctc ccacatccgc
     3781 tctaaccgaa aaggaaggag ttagacaacc tgaagtctag gtccctattt atttttttta
     3841 atagttatgt tagtattaag aacgttattt atatttcaaa tttttctttt ttttctgtac
     3901 aaacgcgtgt acgcatgtaa cattatactg aaaaccttgc ttgagaaggt tttgggacgc
     3961 tcgaaggctt taatttgcaa ggttcgcagt ttacactctc atcgtcgctc tcatcatcgc
     4021 ttccgttgtt gttttcctta gtagcgtctg cttggtaccc agcttcggcc gtatttacag
     4081 ataacgatgt cgttattata tatatatata tatattgtca cttgtaccca gcttttgttc
     4141 cctttagtga gggttaattt cgagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     4201 aaattgttat ccgctcacaa cagctggatg aattgaatag ctttatggac cctgaaacca
     4261 cagccacatt aaccttcttt gatggtcaaa acttatcctt caccataaat atgcctcgca
     4321 aaaaaggtaa ttaacatata tagaattaca ttatttatga aatatcatca ctatctctta
     4381 gcatctttaa tccttttcta catcagataa cttcggtttg ttatcatcgt ctgtattgtc
     4441 atcaattggc gcgcctctcc ctgcaggcat taatgaatcg gccaacgcgc ggggagaggc
     4501 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     4561 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     4621 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     4681 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     4741 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     4801 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     4861 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     4921 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     4981 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     5041 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     5101 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     5161 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     5221 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     5281 ggatctcaag aagatccttt gatcttttgt ttaaacctac ggggtctgac gctcagtgga
     5341 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     5401 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     5461 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     5521 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     5581 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     5641 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     5701 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     5761 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     5821 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     5881 aaaaagcggt tagctccttc ggtcctccga tggttgtcag aagtaagttg gccgcagtgt
     5941 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     6001 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     6061 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     6121 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     6181 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     6241 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     6301 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     6361 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     6421 taggggttcc gcgcacattt ccccgaaaag tgccacct
//
