LOCUS       sequence-265885-        7441 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Integration of ANP1-GFP as an additional copy in genome, use 
            auxotrophic marker URA3(Candida albicans). Marker for yeast early 
            golgi..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7441)
  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 7441)
  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: ZJOM86
FEATURES             Location/Qualifiers
     source          1..7441
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             2553..3269
                     /codon_start=1
                     /product="yeast-enhanced green fluorescent protein"
                     /label=yeGFP
                     /note="codons optimized for translation in S. cerevisiae 
                     and C. albicans"
                     /translation="MSKDEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGIDELYK"
     terminator      3290..3477
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(3345..3369)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     protein_bind    3760..3793
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     terminator      4748..4945
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     protein_bind    4999..5032
                     /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(5086..5105)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5087..5105)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5192..5209)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5363..5951)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5443..5462)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6122..6913,6914..6982))
                     /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     6745..6764
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6983..7087)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7155..7173
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7211..7233)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7333..7352
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        join(7433..7441,1..10)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     join(7433..7441,1..9)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 acactataga acgcggccgc cagctgaaga ttccaatgtc aagatatggc aaaaaccttg
       61 tgcattatat aatcgtggaa cacgacgatc aacgcggcca aaagccgatc gacgacgacg
      121 acgaaaaaaa tttttattac cactgttcct tcaccttcga aacttttttt cgggcgacgg
      181 cgtttctttt agctcctgcg gtatgtgcgg tgcgggaggt cccttgtcgc ctcacacgga
      241 cccgatataa cgctacggaa tacatcgagg ggtatgggtg gatgatatca ctgcagcaag
      301 gtctgggtgt tgcagaattt taccggcctt ggcccctcag cgtccagcta cagcgttata
      361 ctactccaag tcgccgttcc cgttttgctg ttctcacccc tcagagaaaa tgccaccaga
      421 acgaagccaa tggtcaggac ttatcgttac ttatactgtt gagccggata tatccgctgt
      481 gctctaatac cagccaaacg agaagggtag cggcaagaaa gggtaaactt tcatgatgaa
      541 gttagctagt tccttttacg attgctacta tctcgttcaa cttggcctgc gctgttgatg
      601 aaaatagcag tgactgcaga gaatagaaat gaaccgggct catctggtct agttgccttc
      661 ttttttttat tttttttcat tttccttttc ttttcttttc ttttcttgtc tcactatgtt
      721 ggtttaatca attcgtaatc cctttaccaa cgcattgtgt tcaattttcc tcttgtgtgg
      781 ggtcgcccgc cttgtctcat cgtgaaaaaa ttgcgtaaga tgaaatcatg aaaacatgaa
      841 ataaaaaaaa taaataaaaa taactacacg attcaaaaac gccaagatct gttagtagag
      901 acagcaagca ttgcaaaacc attcatcact cagttgctag gcgagtgttg gtgcgattta
      961 ccctcatcat ctttttttcg ctccattaaa cagttccttg tggctttcat gcaaagtgta
     1021 cgtccataat gaagtataat aacagaaaac tctcgttcaa ccctaccaca gtaagtatcg
     1081 ctggaacgtt gcttacggtg ttctttctca caagactcgt gctttcgttc ttctcgatat
     1141 cgctattcca gctggtaact ttccaaggaa tcttcaagcc ctatgttcca gattttaaaa
     1201 atactcccag cgtagagttc tacgacctac gaaattacca aggcaacaaa gatggttggc
     1261 aacagggtga ccgcatcttg ttttgcgtgc cactgagaga tgcttctgag catcttccca
     1321 tgttttttaa ccatctgaat accatgacat atccgcacaa cctgattgat ctgtcgtttt
     1381 tggtgagtga ctcttcggat aacaccatgg gcgtgttact atccaatcta cagatggcgc
     1441 aatcgcaaca agacaagtcc aaaaggtttg gtaacatcga aatctatgag aaggacttcg
     1501 gccaaatcat tggtcagtcc ttttctgatc gtcacgggtt tggcgcccag ggccccagaa
     1561 gaaagctgat ggccagggct cgtaattggc ttgggtccgt ggccctgaaa ccataccact
     1621 cctgggttta ctggagagac gtggatgtcg aaaccatccc cactaccatc atggaggatc
     1681 taatgcatca cgacaaggac gtcattgtcc ccaacgtctg gagaccttta ccagactggc
     1741 tgggaaacat tcaaccctac gatttaaatt catggaagga atcagaaggg ggtcttcaat
     1801 tggcggattc cctagacgaa gatgctgtca ttgtggaagg gtacccagag tacgccactt
     1861 ggagacctca cctggcatac atgagagacc caaacggtaa tccagaagac gaaatggaat
     1921 tggacggtat tggaggggtc tcgattctcg cgaaagctaa agtattcaga acaggctcac
     1981 actttcctgc attctccttt gaaaagcacg cagagacaga agcattcggt aggctctccc
     2041 gcaggatgaa ttacaatgtc atcggcttgc cccattatgt catctggcac atctacgaac
     2101 cttccagcga cgacttaaag cacatggctt ggatggctga agaggagaag cggaaactag
     2161 aagaagagag aattcgcgaa ttttacaaca agatctggga gattggattt gaagatgtca
     2221 gagaccaatg gaacgaagaa agagattcaa tcttaaaaaa cattgactcc actttgaaca
     2281 ataaagtcac tgtagactgg tctgaagagg gagatggctc ggagctggtg gactcaaagg
     2341 gtgactttgt ctcaccaaac aaccaacaac aacaacaaca gcaacaacaa caacaacagc
     2401 agcagcagca acagcaacag caacaacagc tcgacggcaa cccgcagggg aaacctcttg
     2461 atgacaacga caagaacaag aaaaaacatc ctaaagaagt tccattagac ttcgaccctg
     2521 atagaaacct ttctcttgaa gttgctgaag aaatgtctaa agatgaagaa ttattcactg
     2581 gtgttgtccc aattttggtt gaattagatg gtgatgttaa tggtcacaaa ttttctgtct
     2641 ccggtgaagg tgaaggtgat gctacttacg gtaaattgac cttaaaattt atttgtacta
     2701 ctggtaaatt gccagttcca tggccaacct tagtcactac tttaacttat ggtgttcaat
     2761 gtttttctag atacccagat catatgaaac aacatgactt tttcaagtct gccatgccag
     2821 aaggttatgt tcaagaaaga actatttttt tcaaagatga cggtaactac aagaccagag
     2881 ctgaagtcaa gtttgaaggt gataccttag ttaatagaat cgaattaaaa ggtattgatt
     2941 ttaaagaaga tggtaacatt ttaggtcaca aattggaata caactataac tctcacaatg
     3001 tttacatcat ggctgacaaa caaaagaatg gtatcaaagt taacttcaaa attagacaca
     3061 acattgaaga tggttctgtt caattagctg accattatca acaaaatact ccaattggtg
     3121 atggtccagt cttgttacca gacaaccatt acttatccac tcaatctgcc ttatccaaag
     3181 atccaaacga aaagagagac cacatggtct tgttagaatt tgttactgct gctggtatta
     3241 cccatggtat tgatgaattg tacaaataag gcgcgccact tctaaataag cgaatttctt
     3301 atgatttatg atttttatta ttaaataagt tataaaaaaa ataagtgtat acaaatttta
     3361 aagtgactct taggttttaa aacgaaaatt cttattcttg agtaactctt tcctgtaggt
     3421 caggttgctt tctcaggtat agtatgaggt cgctcttatt gaccacacct ctaccggcag
     3481 atccgctagg gataacaggg taatatagat ctgtttagct tgcctcgtcc ccgccgggtc
     3541 acccggccag cgacatggag gcccagaata ccctccttga cagtcttgac gtgcgcagct
     3601 caggggcatg atgtgactgt cgcccgtaca tttagcccat acatccccat gtataatcat
     3661 ttgcatccat acattttgat ggccgcacgg cgcgaagcaa aaattacggc tcctcgctgc
     3721 ggacctgcga gcagggaaac gctcccctca cagacgcgta taacttcgta taatgtatgc
     3781 tatacgaagt tattgaattg tccccacgcc gcgcccctgt agagaaatat aaaaggttag
     3841 gatttgccac tgaggttctt ctttcatata cttcctttta aaatcttgct aggatacagt
     3901 tctcacatca catccgaaca taaacaacca tgacagtcaa cactaagacc tatagtgaga
     3961 gagcagaaac tcatgcctca ccagtagcac aacgattatt tcgattaatg gaactgaaga
     4021 aaaccaattt atgtgcatca attgatgttg ataccactaa ggaattcctt gaattaattg
     4081 ataaattggg tccttatgta tgcttaatca agactcatat tgatataatc aatgattttt
     4141 cctatgaatc cactattgaa ccattattag aactttcacg taaacatcaa tttatgattt
     4201 ttgaagatag aaaatttgct gatattggta ataccgtgaa gaaacaatat attggtggag
     4261 tttataaaat tagtagttgg gcagatatta ctaatgctca tggtgtcact gggaatggag
     4321 tagttgaagg attaaaacag ggagctaaag aaaccaccac caaccaagag ccaagagggt
     4381 tattgatgtt agctgaatta tcatcagtgg gatcattagc atatggagaa tattctcaaa
     4441 aaactgttga aattgctaaa tccgataagg aatttgttat tggatttatt gcccaacgtg
     4501 atatgggtgg acaagaagaa ggatttgatt ggcttattat gacacctgga gttggattag
     4561 atgataaagg tgatggatta ggacaacaat atagaactgt tgatgaagtt gttagcactg
     4621 gaactgatat tatcattgtt ggtagaggat tgtttggtaa aggaagagat ccagatattg
     4681 aaggtaaaag gtatagagat gctggttgga atgcttattt gaaaaagact ggccaattat
     4741 aatcagtact gacaataaaa agattcttgt tttcaagaac ttgtcatttg tatagttttt
     4801 ttatattgta gttgttctat tttaatcaaa tgttagcgtg atttatattt tttttcgcct
     4861 cgacatcatc tgcccagatg cgaagttaag tgcgcagaaa gtaatatcat gcgtcaatcg
     4921 tatgtgaatg ctggtcgcta tactgctgtc gattcgatac taacgccgcc atccagtgtc
     4981 gaaaacgagc tcgaattcat aacttcgtat aatgtatgct atacgaagtt atatcgatga
     5041 tatcagatcc actagtggcc tatgcggccg cggatctgcc ggtctcccta tagtgagtcg
     5101 tattaatttc gataagccag gttaacctgc attaatgaat cggccaacgc gcggggagag
     5161 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     5221 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     5281 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     5341 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     5401 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     5461 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     5521 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     5581 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     5641 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     5701 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     5761 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     5821 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     5881 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     5941 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     6001 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     6061 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     6121 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     6181 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     6241 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     6301 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     6361 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     6421 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     6481 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     6541 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     6601 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     6661 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     6721 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     6781 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     6841 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     6901 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     6961 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     7021 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     7081 ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga
     7141 cattaaccta taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg
     7201 acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg
     7261 atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct
     7321 ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatatg gacatattgt
     7381 cgttagaacg cggctacaat taatacataa ccttatgtat catacacata cgatttaggt
     7441 g
//
