LOCUS       sequence-272374-        7735 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Integration of 2xGFP tag at SEC7 C terminus, use auxotrophic marker 
            URA3(Candida albicans). Marker for yeast late golgi/early endosome..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7735)
  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 7735)
  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: ZJOM156
FEATURES             Location/Qualifiers
     source          1..7735
                     /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             1120..1833
                     /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             1870..2586
                     /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             1870..2583
                     /codon_start=1
                     /product="green fluorescent protein"
                     /label=GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     terminator      2607..2794
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2662..2686)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     protein_bind    3077..3110
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     terminator      4065..4262
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     protein_bind    4316..4349
                     /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(5389..5408)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5390..5408)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5495..5512)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5666..6254)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5746..5765)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6425..7216,7217..7285))
                     /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     7048..7067
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7286..7390)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7458..7476
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7514..7536)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7636..7655
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 atttaggtga cactatagaa cgcggccgcc agctgaagct tgatattgca aaacgtatct
       61 aaattcaatg agtaccattg gaatcaaata ggtgacgtat tcgataaatt gtttgattta
      121 accactgcta atgaattgtt cgattatgat ccgttacagc aaggaagaaa atcatctgta
      181 tctcatcacc aaacaactaa tgataccagc caacactcag atgatgatag caacgacaga
      241 agggagaacg attctaatat cagtgagacg gtcgaaagag cgcatcaaga agaatccagc
      301 gaagacgttg gtggtgacat ggttgaaaca ctcaatgggc aaactaagct taataatggc
      361 aattccgttc cgacggtaaa ggacgaattg aatccaaagc ccgctagtct aagtatcccc
      421 aagaaaacta agcacatgaa acgaaacgaa agtaacgaag atatacgtag gagaataaac
      481 atcaagaatt ctattgttgt caaatgtgtt ttacagcttt tgatgataga gctgctcaac
      541 gaattattcg agaacgaaga ttttgcccac tgtatccctt acaaggaagc cattagaatt
      601 acaagattgt tggagaaatc gtacgaattt tctcgtgact ttaatgaaga ttatgggtta
      661 agaacaagac tagtagaggc tcgtgtagtt gataaaatac caaacctgtt gaaacaagaa
      721 acgagtgctg ctgcagttct ccttgacatt atgtttcagt tgtatctgaa cgatgatgag
      781 aagaaggctg atttaataac tcgtctgatc accatttgca tccaggtcgt agagggttat
      841 gtttctttgg atgatagaac catggaacgt agtatcaatg cctggcgttc agtgatagtt
      901 gaaatattac aaggttacta cgaatttgat gatgaagact ttagactata ctgtcctgct
      961 atgtatgctc tggtgattca aatattagat aagagcgttc caacggagtt gagacatgct
     1021 ataaaacaat ttctaagcag agttggtgaa ttataccttt ctactgatgc ttcgtacgct
     1081 gcaggtcgac ggatcggtga cggtgctggt ttaattaaca tgtctaaagg tgaagaatta
     1141 ttcactggtg ttgtcccaat tttggttgaa ttagatggtg atgttaatgg tcacaaattt
     1201 tctgtctccg gtgaaggtga aggtgatgct acttacggta aattgacctt aaaatttatt
     1261 tgtactactg gtaaattgcc agttccatgg ccaaccttag tcactacttt aacttatggt
     1321 gttcaatgtt tttctagata cccagatcat atgaaacaac atgacttttt caagtctgcc
     1381 atgccagaag gttatgttca agaaagaact atttttttca aagatgacgg taactacaag
     1441 accagagctg aagtcaagtt tgaaggtgat accttagtta atagaatcga attaaaaggt
     1501 attgatttta aagaagatgg taacatttta ggtcacaaat tggaatacaa ctataactct
     1561 cacaatgttt acatcatggc tgacaaacaa aagaatggta tcaaagttaa cttcaaaatt
     1621 agacacaaca ttgaagatgg ttctgttcaa ttagctgacc attatcaaca aaatactcca
     1681 attggtgatg gtccagtctt gttaccagac aaccattact tatccactca atctgcctta
     1741 tccaaagatc caaacgaaaa gagagaccac atggtcttgt tagaatttgt tactgctgct
     1801 ggtattaccc atggtattga tgaattgtac aaaattccag ttccaacgcg tgatctagag
     1861 aagccattca tgtctaaagg tgaagaatta ttcactggtg ttgtcccaat tttggttgaa
     1921 ttagatggtg atgttaatgg tcacaaattt tctgtctccg gtgaaggtga aggtgatgct
     1981 acttacggta aattgacctt aaaatttatt tgtactactg gtaaattgcc agttccatgg
     2041 ccaaccttag tcactacttt aacttatggt gttcaatgtt tttctagata cccagatcat
     2101 atgaaacaac atgacttttt caagtctgcc atgccagaag gttatgttca agaaagaact
     2161 atttttttca aagatgacgg taactacaag accagagctg aagtcaagtt tgaaggtgat
     2221 accttagtta atagaatcga attaaaaggt attgatttta aagaagatgg taacatttta
     2281 ggtcacaaat tggaatacaa ctataactct cacaatgttt acatcatggc tgacaaacaa
     2341 aagaatggta tcaaagttaa cttcaaaatt agacacaaca ttgaagatgg ttctgttcaa
     2401 ttagctgacc attatcaaca aaatactcca attggtgatg gtccagtctt gttaccagac
     2461 aaccattact tatccactca atctgcctta tccaaagatc caaacgaaaa gagagaccac
     2521 atggtcttgt tagaatttgt tactgctgct ggtattaccc atggtatgga tgaattatat
     2581 aaataaggcg cgccacttct aaataagcga atttcttatg atttatgatt tttattatta
     2641 aataagttat aaaaaaaata agtgtataca aattttaaag tgactcttag gttttaaaac
     2701 gaaaattctt attcttgagt aactctttcc tgtaggtcag gttgctttct caggtatagt
     2761 atgaggtcgc tcttattgac cacacctcta ccggcagatc cgctagggat aacagggtaa
     2821 tatagatctg tttagcttgc ctcgtccccg ccgggtcacc cggccagcga catggaggcc
     2881 cagaataccc tccttgacag tcttgacgtg cgcagctcag gggcatgatg tgactgtcgc
     2941 ccgtacattt agcccataca tccccatgta taatcatttg catccataca ttttgatggc
     3001 cgcacggcgc gaagcaaaaa ttacggctcc tcgctgcgga cctgcgagca gggaaacgct
     3061 cccctcacag acgcgtataa cttcgtataa tgtatgctat acgaagttat tgaattgtcc
     3121 ccacgccgcg cccctgtaga gaaatataaa aggttaggat ttgccactga ggttcttctt
     3181 tcatatactt ccttttaaaa tcttgctagg atacagttct cacatcacat ccgaacataa
     3241 acaaccatga cagtcaacac taagacctat agtgagagag cagaaactca tgcctcacca
     3301 gtagcacaac gattatttcg attaatggaa ctgaagaaaa ccaatttatg tgcatcaatt
     3361 gatgttgata ccactaagga attccttgaa ttaattgata aattgggtcc ttatgtatgc
     3421 ttaatcaaga ctcatattga tataatcaat gatttttcct atgaatccac tattgaacca
     3481 ttattagaac tttcacgtaa acatcaattt atgatttttg aagatagaaa atttgctgat
     3541 attggtaata ccgtgaagaa acaatatatt ggtggagttt ataaaattag tagttgggca
     3601 gatattacta atgctcatgg tgtcactggg aatggagtag ttgaaggatt aaaacaggga
     3661 gctaaagaaa ccaccaccaa ccaagagcca agagggttat tgatgttagc tgaattatca
     3721 tcagtgggat cattagcata tggagaatat tctcaaaaaa ctgttgaaat tgctaaatcc
     3781 gataaggaat ttgttattgg atttattgcc caacgtgata tgggtggaca agaagaagga
     3841 tttgattggc ttattatgac acctggagtt ggattagatg ataaaggtga tggattagga
     3901 caacaatata gaactgttga tgaagttgtt agcactggaa ctgatattat cattgttggt
     3961 agaggattgt ttggtaaagg aagagatcca gatattgaag gtaaaaggta tagagatgct
     4021 ggttggaatg cttatttgaa aaagactggc caattataat cagtactgac aataaaaaga
     4081 ttcttgtttt caagaacttg tcatttgtat agttttttta tattgtagtt gttctatttt
     4141 aatcaaatgt tagcgtgatt tatatttttt ttcgcctcga catcatctgc ccagatgcga
     4201 agttaagtgc gcagaaagta atatcatgcg tcaatcgtat gtgaatgctg gtcgctatac
     4261 tgctgtcgat tcgatactaa cgccgccatc cagtgtcgaa aacgagctcg aattcataac
     4321 ttcgtataat gtatgctata cgaagttata tcgatgatat cagatccact agtggcctat
     4381 gcggccgcgg cagattaaaa tatgcttagt tgtagaacac agaaaaaatg agaaggttga
     4441 tatactttta ctcatattaa tcctgattta gtggcggtga tcctttttag attcgtttaa
     4501 ttcataaaca aggtacatca tatagcaata acaaatatgt aaactatgta gaaagactta
     4561 tcaagtttag ccattttggt cctcattcat gggcatttta tactaacttc atttttgccg
     4621 aaataagaaa ggatgtaatc ttaacctaaa atatgtaagc aattaagggc gcacgtagtc
     4681 acttgcgaac aaataggaaa agaaagaaac aagcacggat actgaatgac tgagtgtatt
     4741 actacacgat ttatctcaac gaatttaagt gtggaggacg atcggctgca cgtaggagtc
     4801 aacgtttttt tactgtgttc atcacaaaac cattaagctg agtcatccat attgcaaaca
     4861 ataaggtagc accgcttcca tcaaaacact agaacatacc gatgttttca cggatttatt
     4921 ggaaacaaat agcagatgag ggcgttttta cttgtaatga aggcgaatga gaaggcaagg
     4981 cttacaggct tgttgtttat gccaagcatc aatactcgct atggaggagc gccatcagcg
     5041 ataacaacaa agtttgcaag tgcttgcttg gatataaaag gcttgacgct taagataaat
     5101 agggccatac ttgcttgctt ttactatttt tgttcctcat aagcctgatt gactacgcta
     5161 ttataaaccc ctacattatc ctataatcaa cttcacttgt gtaacgttga atgagttcaa
     5221 ctgagtaata ctcggtcgga aatatacgct ccatctggca ggaaacatag gctgcattta
     5281 tatcttgcac aattatgccc ttcgtaggtg tatgttttca tgttttccat tatgtcaatg
     5341 tagatgtatc tttggacagg caacttttgg ccgcggatct gccggtctcc ctatagtgag
     5401 tcgtattaat ttcgataagc caggttaacc tgcattaatg aatcggccaa cgcgcgggga
     5461 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     5521 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     5581 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     5641 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     5701 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     5761 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     5821 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     5881 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     5941 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     6001 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     6061 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     6121 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     6181 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     6241 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     6301 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     6361 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     6421 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     6481 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     6541 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     6601 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     6661 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     6721 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     6781 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     6841 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     6901 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     6961 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     7021 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     7081 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     7141 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     7201 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     7261 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     7321 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     7381 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca
     7441 tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg
     7501 atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag
     7561 cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg
     7621 gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat atggacatat
     7681 tgtcgttaga acgcggctac aattaataca taaccttatg tatcatacac atacg
//
