LOCUS       sequence-265890-        7898 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Integration of TGL3-GFP as an additional copy in genome, use 
            auxotrophic marker URA3(Candida albicans). Marker for yeast lipid 
            droplet..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7898)
  AUTHORS   Zhu J, Zhang ZT, Tang SW, Zhao BS, Li H, Song JZ, Li D, Xie Z
  TITLE     A Validated Set of Fluorescent-Protein-Based Markers for Major 
            Organelles in Yeast (Saccharomyces cerevisiae).
  JOURNAL   MBio. 2019 Sep 3;10(5). pii: mBio.01691-19. doi: 
            10.1128/mBio.01691-19.
  PUBMED    31481383
REFERENCE   2  (bases 1 to 7898)
  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: "MBio."; 
            date: "2019-09-3"; volume: "10(5). pii: mBio.01691-19. doi"; pages: 
            " 10.1128/mBio.01691-19"
COMMENT     Sequence Label: ZJOM31
FEATURES             Location/Qualifiers
     source          1..7898
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             2370..3086
                     /codon_start=1
                     /product="yeast-enhanced green fluorescent protein"
                     /label=yeGFP
                     /note="codons optimized for translation in S. cerevisiae 
                     and C. albicans"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGIDELYK"
     terminator      3107..3294
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(3162..3186)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     protein_bind    3577..3610
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     terminator      4565..4762
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     protein_bind    4816..4849
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(4903..4922)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4904..4922)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5009..5026)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5180..5768)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5260..5279)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5939..6730,6731..6799))
                     /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     6562..6581
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6800..6904)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6972..6990
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7028..7050)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7150..7169
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        7250..7268
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     7250..7267
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 atatatgaag gcttagttca atgtattctt acccacttct cttgaacttc aattgatcac
       61 cacgtctacc taaatgtagt ggcgtcttgg ttcttttcca tactttgacg atacttttga
      121 atttttaatt ttgccactct ataatagggt atgttcttta taggtcattt ttactttttt
      181 agtagggcat tttaataaat actaggtttt tagtaatttc ggagatactt atcctaggtc
      241 tgaaaattca accctaaccc tttttttctt atgttttttt ttttctgggt agcagctaat
      301 agaaattagt actgattaca tagagaagta ctatgacaca atagtaaggg aatcatctat
      361 tcatatatca catctttgag ttgccgttaa gcatgaagga aacggcgcag gaatacaagg
      421 tgtctgctgt aataccgacc cttttgaaaa actggatact gcgtgtagtg tatgccacat
      481 tggaccatat acctcctttt gtgtgggaaa ttctacatgt catcaccgat atttatttct
      541 tctgggttca aaagctgatt aactatgtgc ggccacactc aagagtcatt tattataacg
      601 ctattaagaa gttagatgag tgtgatacgt atcaaatgtg gtgtcagcaa gcgtccgtag
      661 tggatgaaat aacaggcgca aatttatggc ggcggaattt cttttcgagg aggtacgact
      721 tcaattctgt cattgaacaa tatagcatat tggaaaatat gttaagggag gagaagtatg
      781 atgtagttaa ggagaagttt tcgactacag gcccttgtat gttgaggaac tttgcgggta
      841 ttggagataa aaagttgttc acaaaatctt tgatgggtac aaagctactt attgaacaat
      901 atttgacccg aattctagaa ggcttggata tattaaataa tcaaacttta actccaacat
      961 cctttttcca aagatgtaag ttgtcactgg gcacaacagc attaatcttg caaggtggtt
     1021 cattgtttgg gctgtttcac cttggggtga ttagaggttt attgttacag gacctgatgc
     1081 ctaatattat aagtggtagc tctatggggg catgcgttgc aagtttattc ggttgcttgt
     1141 ccaatgagca actaaagcaa ttattgaccg atgacaacct cttgaatatc atcaagaatg
     1201 atgtagattt attaaaaagc tgtggatacg gtaatttaga gcagcaccta aacctgggga
     1261 ctttgattca gaacctaata caccacggtt attctcaaga cgtttatctt tttatccggt
     1321 ttgttatgaa atatattgtt aaggaaaaaa cttttgaaga agtttatcaa attactggaa
     1381 aagttttcaa tattgttatt catccaactg ataaatcatg ccctaattta ctaaattatg
     1441 tgacaacccc aaacgtatta atcaagtcag ccattgaatg ctcacttggt tctggtgtaa
     1501 tttcagagga tacatcacta ttatgtaaaa atttagaaaa tgaaattgag ccttttctta
     1561 acatcaataa aaataaacaa gtcaaatttc ttaccccaga aaatgccaac aatccgagca
     1621 taacggagag cccctatact aggttaacgg aactattcaa tgttaataac ttcattgttt
     1681 ctttggcgag accatacttg gcaccattgg tggtaaatga tttgaagcat gaaataaaaa
     1741 cgtcaaaata ttactattat aagcattatc caaatatgcc tcctataaac gcgaatacag
     1801 taagaaaaac acagagatcc tcttcgcaat cacccataaa ggcaggaact gtcgaggatc
     1861 ttgagccaga gcctttaatg agccctgtgc ccccaagttc ggcagtcaac gattcagcag
     1921 agtacataat cccagaattg ggcataccgc agttgaactt tactgaaatg gaaccgctgg
     1981 cgtttaaatt taagtatcat ctggaaagaa aattaaaaaa tattgccact atggaatttc
     2041 gacataggat ggaggtattg gataatttag gcctgttgtg ctcattaata aaaagactaa
     2101 taattgatga aaagactccc agatcagcta cggagattgc tgtggtacca aggatgaaaa
     2161 gtctatcatt aacaaggatt attgaaggtc aactaaacaa cattccatac tggataaaat
     2221 ctggtgaacg aagtacatgg cccgcactag ctttaattaa gacaagatgt gcagtcgaat
     2281 ttaaattaga cgacataata agagcaagac ggagtagggc ttcgtacgct gcaggtcgac
     2341 ggatcggtga cggtgctggt ttaattaaca tgtctaaagg tgaagaatta ttcactggtg
     2401 ttgtcccaat tttggttgaa ttagatggtg atgttaatgg tcacaaattt tctgtctccg
     2461 gtgaaggtga aggtgatgct acttacggta aattgacctt aaaatttatt tgtactactg
     2521 gtaaattgcc agttccatgg ccaaccttag tcactacttt aacttatggt gttcaatgtt
     2581 tttctagata cccagatcat atgaaacaac atgacttttt caagtctgcc atgccagaag
     2641 gttatgttca agaaagaact atttttttca aagatgacgg taactacaag accagagctg
     2701 aagtcaagtt tgaaggtgat accttagtta atagaatcga attaaaaggt attgatttta
     2761 aagaagatgg taacatttta ggtcacaaat tggaatacaa ctataactct cacaatgttt
     2821 acatcatggc tgacaaacaa aagaatggta tcaaagttaa cttcaaaatt agacacaaca
     2881 ttgaagatgg ttctgttcaa ttagctgacc attatcaaca aaatactcca attggtgatg
     2941 gtccagtctt gttaccagac aaccattact tatccactca atctgcctta tccaaagatc
     3001 caaacgaaaa gagagaccac atggtcttgt tagaatttgt tactgctgct ggtattaccc
     3061 atggtattga tgaattgtac aaataaggcg cgccacttct aaataagcga atttcttatg
     3121 atttatgatt tttattatta aataagttat aaaaaaaata agtgtataca aattttaaag
     3181 tgactcttag gttttaaaac gaaaattctt attcttgagt aactctttcc tgtaggtcag
     3241 gttgctttct caggtatagt atgaggtcgc tcttattgac cacacctcta ccggcagatc
     3301 cgctagggat aacagggtaa tatagatctg tttagcttgc ctcgtccccg ccgggtcacc
     3361 cggccagcga catggaggcc cagaataccc tccttgacag tcttgacgtg cgcagctcag
     3421 gggcatgatg tgactgtcgc ccgtacattt agcccataca tccccatgta taatcatttg
     3481 catccataca ttttgatggc cgcacggcgc gaagcaaaaa ttacggctcc tcgctgcgga
     3541 cctgcgagca gggaaacgct cccctcacag acgcgtataa cttcgtataa tgtatgctat
     3601 acgaagttat tgaattgtcc ccacgccgcg cccctgtaga gaaatataaa aggttaggat
     3661 ttgccactga ggttcttctt tcatatactt ccttttaaaa tcttgctagg atacagttct
     3721 cacatcacat ccgaacataa acaaccatga cagtcaacac taagacctat agtgagagag
     3781 cagaaactca tgcctcacca gtagcacaac gattatttcg attaatggaa ctgaagaaaa
     3841 ccaatttatg tgcatcaatt gatgttgata ccactaagga attccttgaa ttaattgata
     3901 aattgggtcc ttatgtatgc ttaatcaaga ctcatattga tataatcaat gatttttcct
     3961 atgaatccac tattgaacca ttattagaac tttcacgtaa acatcaattt atgatttttg
     4021 aagatagaaa atttgctgat attggtaata ccgtgaagaa acaatatatt ggtggagttt
     4081 ataaaattag tagttgggca gatattacta atgctcatgg tgtcactggg aatggagtag
     4141 ttgaaggatt aaaacaggga gctaaagaaa ccaccaccaa ccaagagcca agagggttat
     4201 tgatgttagc tgaattatca tcagtgggat cattagcata tggagaatat tctcaaaaaa
     4261 ctgttgaaat tgctaaatcc gataaggaat ttgttattgg atttattgcc caacgtgata
     4321 tgggtggaca agaagaagga tttgattggc ttattatgac acctggagtt ggattagatg
     4381 ataaaggtga tggattagga caacaatata gaactgttga tgaagttgtt agcactggaa
     4441 ctgatattat cattgttggt agaggattgt ttggtaaagg aagagatcca gatattgaag
     4501 gtaaaaggta tagagatgct ggttggaatg cttatttgaa aaagactggc caattataat
     4561 cagtactgac aataaaaaga ttcttgtttt caagaacttg tcatttgtat agttttttta
     4621 tattgtagtt gttctatttt aatcaaatgt tagcgtgatt tatatttttt ttcgcctcga
     4681 catcatctgc ccagatgcga agttaagtgc gcagaaagta atatcatgcg tcaatcgtat
     4741 gtgaatgctg gtcgctatac tgctgtcgat tcgatactaa cgccgccatc cagtgtcgaa
     4801 aacgagctcg aattcataac ttcgtataat gtatgctata cgaagttata tcgatgatat
     4861 cagatccact agtggcctat gcggccgcgg atctgccggt ctccctatag tgagtcgtat
     4921 taatttcgat aagccaggtt aacctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     4981 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     5041 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     5101 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     5161 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     5221 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     5281 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     5341 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     5401 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     5461 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     5521 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     5581 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     5641 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     5701 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     5761 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     5821 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     5881 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     5941 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     6001 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     6061 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     6121 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     6181 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     6241 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     6301 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     6361 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     6421 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     6481 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     6541 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     6601 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     6661 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     6721 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     6781 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     6841 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     6901 cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat
     6961 taacctataa aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg
     7021 gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg
     7081 ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc
     7141 ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatggac atattgtcgt
     7201 tagaacgcgg ctacaattaa tacataacct tatgtatcat acacatacga tttaggtgac
     7261 actatagaac gcggccgcca gctgaagctt gactggctga tagcctcgac gccagctttg
     7321 attagttcat ccgggttccc gtcaattttt ataaacgtgt ctaaagttct ctctagataa
     7381 gtctttgccc cttggcttct tgcgccaata gctgtaccat atagttctgt gacatttccg
     7441 gaaggttgga attctaaaag atgagcaccg ctcttgtcat acccgatgat caacaaacca
     7501 acaccgtagg gtctgccccc ataggattgt gtatttttct gtgctttatc gcaaagcaaa
     7561 tgacctgctc tctcgacggc aagctttctg ttaaaaacta agctggagta attacactgt
     7621 tgtctcaaat aattacttaa aactctagca tctggagcca acccggctaa cgagagcccc
     7681 atgtgctcgt cacatttgat gatttttttc tggtatgatg acaattcgtc tgcatttctt
     7741 ttcaacgcta ctaaaacagc atgagtgttc gaacgtaacc ccacagtgac gctcccttgt
     7801 ttaatagctt ccaaggcgta ttccacttgg aacagacggc ctgtaggaga aaaagtgact
     7861 gtgtccccgt cgtaattgtt cctgaacatg ttttgtaa
//
