LOCUS       sequence-265864-        6692 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Integration of SNF7-GFP as an additional copy in genome, use 
            auxotrophic marker URA3(Candida albicans). Present on endosomes and 
            vacuolar surface. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6692)
  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 6692)
  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: ZJOM8
FEATURES             Location/Qualifiers
     source          1..6692
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..19
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1..18
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             1813..2529
                     /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      2550..2737
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2605..2629)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     protein_bind    3020..3053
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     terminator      4008..4205
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     protein_bind    4259..4292
                     /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(4346..4365)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4347..4365)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4452..4469)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4623..5211)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4703..4722)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5382..6173,6174..6242))
                     /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     6005..6024
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6243..6347)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6415..6433
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6471..6493)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6593..6612
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 atttaggtga cactatagaa cgcggccgcc agctgaagct tatgcgttga ttattgggtt
       61 tctccccaca gctgtacatt gacgatgctc tttattgata tcgcctaaaa gttcaacaaa
      121 cgccactgga aattgccgcc gttgtcacca caacgtgata agtgatggtg tgctttttgc
      181 acatctagtg aagaaacggg caattaggat ctaggaccag taacacttgc cccgcaaata
      241 ttataacatg ggccttatga ttgctcggtt tgttacatct aacatcctaa gaagtgagtg
      301 gcatgaatac gtagctatat gtagattgaa aattgatgag tcgagtcatc accttcctct
      361 aaacggggga taaattttgg cggtattttc cctaatttta ttcttcgtta tgtcagccat
      421 gtgagactaa ttaattcaag ttttgactta caattgcggc ttcgtcggaa atcagtatta
      481 cgtataaact gtgtaccata aaattggcgt actcggtatt gtttatacta agccggccat
      541 tctagtgatt tcgcctctat aaatggttgt tattcgagta taatatttca accgcatcaa
      601 agaaagaggt agatagcttg cgttcaacac cgttcttttc tttccccctt aatttagcgc
      661 ttggcggtta tgctatggtg acagttaact tttttaatta taggatgact tgtcatattg
      721 ttatatgccg catacttatg taaagtagtt attgaggtgg gggtgagcag agtaacgaca
      781 acgatggagt tccgtaacta accaatgctt aatgttggta agtaatagca aatatatata
      841 ctacatagct tttgttatat gcatcatttt atacgattca tgttcgaccc ttttgataaa
      901 agatgggatt tttaaataag tcggtgggtg atttcgaatc atgaatagtt ggcagagcag
      961 gggaagtacg agcttctaaa gggtaagata ttgtatttcg gacggaagca gcagaaacat
     1021 aacagtattg ataaataagg catgtggtca tcactttttg gttggacatc aagtaatgcc
     1081 aagaataaag agtcaccaac aaaggccata gtgcggttga gggagcatat caaccttcta
     1141 tccaaaaagc aatcgcattt acgtactcaa attacaaatc aagagaatga agctagaatc
     1201 tttttgacga agggcaataa agtaatggcg aagaatgcac ttaaaaagaa gaagacaatc
     1261 gaacaacttt taagtaaggt agaaggcaca atggagtcta tggaacagca gctattctct
     1321 atagaaagtg caaacttaaa tctagagaca atgagggcta tgcaggaagg tgctaaggca
     1381 atgaaaacta ttcacagtgg ccttgacata gataaagtgg atgaaactat ggacgagata
     1441 agggagcaag tcgaattagg agatgaaata agcgacgcta tatccaggcc tttaattact
     1501 ggggcaaacg aggtggatga agatgagctg gacgaggaat tggacatgct ggctcaagaa
     1561 aatgctaacc aagaaacgtc caagatcgtt aataataatg ttaatgcggc gcctatctca
     1621 gagaacaaag tctcactacc tagtgttcca agtaataaaa ttaaacaaag tgagaactct
     1681 gtgaaggacg gggaagagga agaggatgaa gaagatgaag atgaaaaagc attaagagaa
     1741 ctacaagcag aaatggggct tgcttcgtac gctgcaggtc gacggatcgg tgacggtgct
     1801 ggtttaatta acatgtctaa aggtgaagaa ttattcactg gtgttgtccc aattttggtt
     1861 gaattagatg gtgatgttaa tggtcacaaa ttttctgtct ccggtgaagg tgaaggtgat
     1921 gctacttacg gtaaattgac cttaaaattt atttgtacta ctggtaaatt gccagttcca
     1981 tggccaacct tagtcactac tttaacttat ggtgttcaat gtttttctag atacccagat
     2041 catatgaaac aacatgactt tttcaagtct gccatgccag aaggttatgt tcaagaaaga
     2101 actatttttt tcaaagatga cggtaactac aagaccagag ctgaagtcaa gtttgaaggt
     2161 gataccttag ttaatagaat cgaattaaaa ggtattgatt ttaaagaaga tggtaacatt
     2221 ttaggtcaca aattggaata caactataac tctcacaatg tttacatcat ggctgacaaa
     2281 caaaagaatg gtatcaaagt taacttcaaa attagacaca acattgaaga tggttctgtt
     2341 caattagctg accattatca acaaaatact ccaattggtg atggtccagt cttgttacca
     2401 gacaaccatt acttatccac tcaatctgcc ttatccaaag atccaaacga aaagagagac
     2461 cacatggtct tgttagaatt tgttactgct gctggtatta cccatggtat tgatgaattg
     2521 tacaaataag gcgcgccact tctaaataag cgaatttctt atgatttatg atttttatta
     2581 ttaaataagt tataaaaaaa ataagtgtat acaaatttta aagtgactct taggttttaa
     2641 aacgaaaatt cttattcttg agtaactctt tcctgtaggt caggttgctt tctcaggtat
     2701 agtatgaggt cgctcttatt gaccacacct ctaccggcag atccgctagg gataacaggg
     2761 taatatagat ctgtttagct tgcctcgtcc ccgccgggtc acccggccag cgacatggag
     2821 gcccagaata ccctccttga cagtcttgac gtgcgcagct caggggcatg atgtgactgt
     2881 cgcccgtaca tttagcccat acatccccat gtataatcat ttgcatccat acattttgat
     2941 ggccgcacgg cgcgaagcaa aaattacggc tcctcgctgc ggacctgcga gcagggaaac
     3001 gctcccctca cagacgcgta taacttcgta taatgtatgc tatacgaagt tattgaattg
     3061 tccccacgcc gcgcccctgt agagaaatat aaaaggttag gatttgccac tgaggttctt
     3121 ctttcatata cttcctttta aaatcttgct aggatacagt tctcacatca catccgaaca
     3181 taaacaacca tgacagtcaa cactaagacc tatagtgaga gagcagaaac tcatgcctca
     3241 ccagtagcac aacgattatt tcgattaatg gaactgaaga aaaccaattt atgtgcatca
     3301 attgatgttg ataccactaa ggaattcctt gaattaattg ataaattggg tccttatgta
     3361 tgcttaatca agactcatat tgatataatc aatgattttt cctatgaatc cactattgaa
     3421 ccattattag aactttcacg taaacatcaa tttatgattt ttgaagatag aaaatttgct
     3481 gatattggta ataccgtgaa gaaacaatat attggtggag tttataaaat tagtagttgg
     3541 gcagatatta ctaatgctca tggtgtcact gggaatggag tagttgaagg attaaaacag
     3601 ggagctaaag aaaccaccac caaccaagag ccaagagggt tattgatgtt agctgaatta
     3661 tcatcagtgg gatcattagc atatggagaa tattctcaaa aaactgttga aattgctaaa
     3721 tccgataagg aatttgttat tggatttatt gcccaacgtg atatgggtgg acaagaagaa
     3781 ggatttgatt ggcttattat gacacctgga gttggattag atgataaagg tgatggatta
     3841 ggacaacaat atagaactgt tgatgaagtt gttagcactg gaactgatat tatcattgtt
     3901 ggtagaggat tgtttggtaa aggaagagat ccagatattg aaggtaaaag gtatagagat
     3961 gctggttgga atgcttattt gaaaaagact ggccaattat aatcagtact gacaataaaa
     4021 agattcttgt tttcaagaac ttgtcatttg tatagttttt ttatattgta gttgttctat
     4081 tttaatcaaa tgttagcgtg atttatattt tttttcgcct cgacatcatc tgcccagatg
     4141 cgaagttaag tgcgcagaaa gtaatatcat gcgtcaatcg tatgtgaatg ctggtcgcta
     4201 tactgctgtc gattcgatac taacgccgcc atccagtgtc gaaaacgagc tcgaattcat
     4261 aacttcgtat aatgtatgct atacgaagtt atatcgatga tatcagatcc actagtggcc
     4321 tatgcggccg cggatctgcc ggtctcccta tagtgagtcg tattaatttc gataagccag
     4381 gttaacctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     4441 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     4501 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     4561 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     4621 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     4681 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     4741 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     4801 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     4861 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     4921 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     4981 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     5041 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     5101 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     5161 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     5221 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     5281 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     5341 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     5401 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     5461 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     5521 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     5581 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     5641 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     5701 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     5761 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     5821 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     5881 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     5941 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     6001 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     6061 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     6121 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     6181 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     6241 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     6301 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     6361 aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg
     6421 cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac
     6481 atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc
     6541 cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca
     6601 gagcagattg tactgagagt gcaccatatg gacatattgt cgttagaacg cggctacaat
     6661 taatacataa ccttatgtat catacacata cg
//
