LOCUS       sequence-267535-        7808 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Integration of MSP1-2GFP as an additional copy in genome, use 
            auxotrophic marker URA3(Candida albicans). Present on mitochondria 
            and peroxisomes. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7808)
  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 7808)
  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: ZJOM25
FEATURES             Location/Qualifiers
     source          1..7808
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             2158..2871
                     /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"
     CDS             2908..3624
                     /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
                     FVTAAGITHGMDELYK"
     CDS             2908..3621
                     /codon_start=1
                     /product="green fluorescent protein"
                     /label=GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     terminator      3645..3832
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(3700..3724)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     protein_bind    4115..4148
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     terminator      5103..5300
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     protein_bind    5354..5387
                     /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(5441..5460)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5442..5460)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5547..5564)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5718..6306)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5798..5817)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6477..7268,7269..7337))
                     /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     7100..7119
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7338..7442)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7510..7528
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7566..7588)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7688..7707
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        7788..7806
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     7788..7805
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 gcggccgcca gctgaagctt gaagtatcag gaaatcatct gcacgtatgc tatatcgaaa
       61 ccaaaaagaa tattgcagac gtaatgacca aacctcttcc gataaaaaca ttcaaactat
      121 taacaaacaa atggattcat tagatctatt acattatggg tggtatgttg gaataaaaat
      181 ccactatcgt ctatcaacta atagttatat tatcaatata ttatcatata cggtgttaag
      241 atgatgacat aagttatgag aagctgtcat cgaagttaga ggaagctgaa acgcaaggat
      301 tgataatgta ataggatcaa tgaatataaa catataaaac ggaatgagga ataatcgtaa
      361 tattagtatg tagaaatata gattccattt tgaggattcc tatatcctcg aggagaactt
      421 ctagtatatt ctgtatacct aatattatag cctttatcaa caatggaatc ccaacaatta
      481 tctcaacatt cacatatttc tcattctcaa tcttcatggt atctttttta caccgtatat
      541 gataaaatac aagtaatata aataccagcc actggatgac attgaaattt tattctaata
      601 aatgcatcag ctctttaaaa acattgctat ttagctattt gaagccagaa attttcacag
      661 attaccatcc gatatagtgt aacggctatc acatcacgct ttcaccgtgg agaccggggt
      721 tcgactcccc gtatcggagt gttctttttc ttacctcttg aaaaagaggt agtaaagtca
      781 atataaattc ttggctgaag tatttcaata caattggtca aggaagttat tattttttta
      841 atgacttata tctgtccaac acaacaggtt acaccatata atctctattt actggacagg
      901 cggaaatcga tagtttagct ttcctaaaaa tcgttatatt atagaagcgt cttgacacgt
      961 tttcaaaaaa ggaagaagca agaacgaaaa gagataagga ttcaaaagaa aggaagccca
     1021 atgtctcgca aatttgattt aaaaacgatt actgatcttt ccgtcctagt tgggactggc
     1081 atatcattat actacctggt cagtcggcta ctcaacgatg ttgaatcagg accgttatca
     1141 ggtaaatcaa gggaatcaaa ggccaaacag tccttgcaat gggaaaagct ggtgaaaaga
     1201 tcaccggcat tggcagaagt aacattagat gcatatgaaa ggactatatt atcttctatc
     1261 gttacgccag atgaaataaa tattacattc caggatattg gtggcttaga cccgcttatt
     1321 tcagatctac atgaaagtgt catatatccc ttgatgatgc cagaagtata ttctaatagc
     1381 cctttacttc aagcacctag cggtgtcttg ctatatgggc caccaggatg tggtaaaacc
     1441 atgttggcga aggccctagc caaggaaagt ggtgctaatt ttatctcaat aagaatgtca
     1501 tctataatgg ataaatggta tggtgaatct aacaaaatag tcgatgcaat gttttcattg
     1561 gcgaacaagt tacaaccttg tataatattc attgacgaaa ttgattcatt ccttagagaa
     1621 cggtcttcta cagatcatga agttacggca accttaaaag ctgaattcat gactttatgg
     1681 gatggcttat tgaataatgg aagggttatg attatcggtg ctactaatcg gataaatgac
     1741 atagatgatg cgtttttgag gaggttacca aaaaggtttc ttgtttcatt gcctggttct
     1801 gatcaacgtt acaaaatatt aagtgtttta ttaaaagaca ctaaactaga cgaagacgaa
     1861 ttcgacttgc aactgattgc agacaatacc aaaggatttt ctgggtcgga cctgaaagag
     1921 ctttgcagag aagcggcctt agatgcagca aaggaataca taaaacagaa gaggcagctc
     1981 attgatagtg gtacaatcga tgttaatgat acttcttcct tgaagataag accattaaag
     2041 acaaaagatt ttacaaaaaa attaagaatg gatgctacaa gtacgttgtc atctcaacct
     2101 cttgatgctt cgtacgctgc aggtcgacgg atcggtgacg gtgctggttt aattaacatg
     2161 tctaaaggtg aagaattatt cactggtgtt gtcccaattt tggttgaatt agatggtgat
     2221 gttaatggtc acaaattttc tgtctccggt gaaggtgaag gtgatgctac ttacggtaaa
     2281 ttgaccttaa aatttatttg tactactggt aaattgccag ttccatggcc aaccttagtc
     2341 actactttaa cttatggtgt tcaatgtttt tctagatacc cagatcatat gaaacaacat
     2401 gactttttca agtctgccat gccagaaggt tatgttcaag aaagaactat ttttttcaaa
     2461 gatgacggta actacaagac cagagctgaa gtcaagtttg aaggtgatac cttagttaat
     2521 agaatcgaat taaaaggtat tgattttaaa gaagatggta acattttagg tcacaaattg
     2581 gaatacaact ataactctca caatgtttac atcatggctg acaaacaaaa gaatggtatc
     2641 aaagttaact tcaaaattag acacaacatt gaagatggtt ctgttcaatt agctgaccat
     2701 tatcaacaaa atactccaat tggtgatggt ccagtcttgt taccagacaa ccattactta
     2761 tccactcaat ctgccttatc caaagatcca aacgaaaaga gagaccacat ggtcttgtta
     2821 gaatttgtta ctgctgctgg tattacccat ggtattgatg aattgtacaa aattccagtt
     2881 ccaacgcgtg atctagagaa gccattcatg tctaaaggtg aagaattatt cactggtgtt
     2941 gtcccaattt tggttgaatt agatggtgat gttaatggtc acaaattttc tgtctccggt
     3001 gaaggtgaag gtgatgctac ttacggtaaa ttgaccttaa aatttatttg tactactggt
     3061 aaattgccag ttccatggcc aaccttagtc actactttaa cttatggtgt tcaatgtttt
     3121 tctagatacc cagatcatat gaaacaacat gactttttca agtctgccat gccagaaggt
     3181 tatgttcaag aaagaactat ttttttcaaa gatgacggta actacaagac cagagctgaa
     3241 gtcaagtttg aaggtgatac cttagttaat agaatcgaat taaaaggtat tgattttaaa
     3301 gaagatggta acattttagg tcacaaattg gaatacaact ataactctca caatgtttac
     3361 atcatggctg acaaacaaaa gaatggtatc aaagttaact tcaaaattag acacaacatt
     3421 gaagatggtt ctgttcaatt agctgaccat tatcaacaaa atactccaat tggtgatggt
     3481 ccagtcttgt taccagacaa ccattactta tccactcaat ctgccttatc caaagatcca
     3541 aacgaaaaga gagaccacat ggtcttgtta gaatttgtta ctgctgctgg tattacccat
     3601 ggtatggatg aattatataa ataaggcgcg ccacttctaa ataagcgaat ttcttatgat
     3661 ttatgatttt tattattaaa taagttataa aaaaaataag tgtatacaaa ttttaaagtg
     3721 actcttaggt tttaaaacga aaattcttat tcttgagtaa ctctttcctg taggtcaggt
     3781 tgctttctca ggtatagtat gaggtcgctc ttattgacca cacctctacc ggcagatccg
     3841 ctagggataa cagggtaata tagatctgtt tagcttgcct cgtccccgcc gggtcacccg
     3901 gccagcgaca tggaggccca gaataccctc cttgacagtc ttgacgtgcg cagctcaggg
     3961 gcatgatgtg actgtcgccc gtacatttag cccatacatc cccatgtata atcatttgca
     4021 tccatacatt ttgatggccg cacggcgcga agcaaaaatt acggctcctc gctgcggacc
     4081 tgcgagcagg gaaacgctcc cctcacagac gcgtataact tcgtataatg tatgctatac
     4141 gaagttattg aattgtcccc acgccgcgcc cctgtagaga aatataaaag gttaggattt
     4201 gccactgagg ttcttctttc atatacttcc ttttaaaatc ttgctaggat acagttctca
     4261 catcacatcc gaacataaac aaccatgaca gtcaacacta agacctatag tgagagagca
     4321 gaaactcatg cctcaccagt agcacaacga ttatttcgat taatggaact gaagaaaacc
     4381 aatttatgtg catcaattga tgttgatacc actaaggaat tccttgaatt aattgataaa
     4441 ttgggtcctt atgtatgctt aatcaagact catattgata taatcaatga tttttcctat
     4501 gaatccacta ttgaaccatt attagaactt tcacgtaaac atcaatttat gatttttgaa
     4561 gatagaaaat ttgctgatat tggtaatacc gtgaagaaac aatatattgg tggagtttat
     4621 aaaattagta gttgggcaga tattactaat gctcatggtg tcactgggaa tggagtagtt
     4681 gaaggattaa aacagggagc taaagaaacc accaccaacc aagagccaag agggttattg
     4741 atgttagctg aattatcatc agtgggatca ttagcatatg gagaatattc tcaaaaaact
     4801 gttgaaattg ctaaatccga taaggaattt gttattggat ttattgccca acgtgatatg
     4861 ggtggacaag aagaaggatt tgattggctt attatgacac ctggagttgg attagatgat
     4921 aaaggtgatg gattaggaca acaatataga actgttgatg aagttgttag cactggaact
     4981 gatattatca ttgttggtag aggattgttt ggtaaaggaa gagatccaga tattgaaggt
     5041 aaaaggtata gagatgctgg ttggaatgct tatttgaaaa agactggcca attataatca
     5101 gtactgacaa taaaaagatt cttgttttca agaacttgtc atttgtatag tttttttata
     5161 ttgtagttgt tctattttaa tcaaatgtta gcgtgattta tatttttttt cgcctcgaca
     5221 tcatctgccc agatgcgaag ttaagtgcgc agaaagtaat atcatgcgtc aatcgtatgt
     5281 gaatgctggt cgctatactg ctgtcgattc gatactaacg ccgccatcca gtgtcgaaaa
     5341 cgagctcgaa ttcataactt cgtataatgt atgctatacg aagttatatc gatgatatca
     5401 gatccactag tggcctatgc ggccgcggat ctgccggtct ccctatagtg agtcgtatta
     5461 atttcgataa gccaggttaa cctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     5521 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     5581 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     5641 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     5701 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     5761 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     5821 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     5881 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     5941 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     6001 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     6061 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     6121 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     6181 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     6241 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     6301 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     6361 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     6421 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     6481 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     6541 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     6601 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     6661 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     6721 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     6781 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     6841 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     6901 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     6961 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     7021 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     7081 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     7141 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     7201 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     7261 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     7321 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     7381 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     7441 cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta
     7501 acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt
     7561 gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc
     7621 gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt
     7681 aactatgcgg catcagagca gattgtactg agagtgcacc atatggacat attgtcgtta
     7741 gaacgcggct acaattaata cataacctta tgtatcatac acatacgatt taggtgacac
     7801 tatagaac
//
