LOCUS       sequence-267441-        7985 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Integration of CHS5-GFP as an additional copy in genome, 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 7985)
  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 7985)
  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: ZJOM5
FEATURES             Location/Qualifiers
     source          1..7985
                     /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             3106..3822
                     /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      3843..4030
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(3898..3922)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     protein_bind    4313..4346
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     terminator      5301..5498
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     protein_bind    5552..5585
                     /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(5639..5658)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5640..5658)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5745..5762)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5916..6504)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5996..6015)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6675..7466,7467..7535))
                     /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     7298..7317
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7536..7640)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7708..7726
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7764..7786)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7886..7905
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 atttaggtga cactatagaa cgcggccgcc agctgaagct tctaactata ataactggta
       61 gctttgtcac tcgtaccagg aaaagtgaag attaaactga attttaaaat gaacgatcga
      121 ttagttacgg aagagcaaga gttgtttaca aaattgcgcg agattgtagg ttcaagtatt
      181 cgcttttggg aggaacaact gttttatcaa gttcaagatg taagcaccat agaaaaccac
      241 gtcattctca gtttaaaatg tacaatttta acggatgctc agataagtac gttcataagc
      301 aaacccagag agcttcatac gcatgccaaa ggatatcctg aaatctatta cctttccgag
      361 ttatcaacaa ctgtcaattt tttttctaaa gagggaaact atgtcgaaat aagccaggtt
      421 attcctcatt ttaatgaata tttttcctct ttaatagtgt ctcaattgga atttgaatac
      481 ccgatggtct tctccatgat ttcaaggctc cgattgaagt ggcaacaaag ttcgctcgct
      541 ccgatatcct acgccctaac gagcaattca gtacttcttc caataatgct taacatgatt
      601 gcccaagaca aatcttcaac aaccgcgtat caaattctgt gtcgaagaag aggtcctcca
      661 attcagaatt ttcaaatttt ttccttaccg gctgtaacgt acaataagta gcatgcataa
      721 aatataattt aatcaaatac ttttgggcaa ttaaaatttt agttaacaat agttatgcaa
      781 tgcgctttat gttcatatga taccgtttat aagctattgc catatcctta tcttattgct
      841 tccagtagcc tcgagtcgac cactaaaaag atgtcactta agacggaaat tatgtagctg
      901 cacttctttt ttaacaagtt cggtcggccc ttcaagttct cctttctaaa gcctcattat
      961 ttattgcgta gatgctaaat gttatcgcgg tttagcttgc atgttacgtt tccgttttag
     1021 aacctggtcg agtagcgaat aatgtcttca gttgatgtac tgttaacagt aggtaagttg
     1081 gatgcctcat tggcgttact gactactcag gatcatcatg ttattgagtt tcctacagta
     1141 ttattaccag aaaatgttaa agctggatct atcataaaaa tgcaagtttc acaaaattta
     1201 gaggaggaaa aaaaacaaag gaatcatttt aagagtatac aagccaaaat tttggagaag
     1261 tatggtaccc ataaaccgga gagcccagtt ttgaaaattg ttaacgttac gcaaacgagc
     1321 tgtgttctag catgggatcc attgaaactt ggctcagcaa aattgaaatc actgatcctt
     1381 tataggaagg gaatacgttc aatggtaatt ccaaatccat tcaaagtgac taccacgaaa
     1441 atatccggtc tttccgttga tacgccatac gaatttcaat tgaaactgat aaccacgtca
     1501 ggaacattat ggtctgaaaa ggttatattg cgtacacata agatgactga catgtctggt
     1561 atcactgtat gtttgggtcc attggatcca ttgaaagaaa tttcagactt acagatatcc
     1621 caatgtttgt ctcacatcgg ggcgagacct ttacaacgtc atgttgcgat agatactacg
     1681 cattttgtct gtaacgatct agacaatgaa gaaagcaatg aagagcttat aagggcaaaa
     1741 cataacaaca taccaattgt cagaccggaa tgggtgagag cttgtgaggt tgagaaaaga
     1801 atcgttggtg ttagaggatt ttacttagat gcagatcaaa gtatactgaa aaactacaca
     1861 ttcccaccag ttaatgagga agaactttcg tactcaaagg agaatgagcc ggtagccgaa
     1921 gtagcggatg aaaataagat gcccgaggac acaacagatg tcgaacaggt tgcatcacct
     1981 aatgacaatg agagtaatcc ttcagaagct aaggaacaag gagaaaagag tggacatgaa
     2041 actgccccag taagtcctgt agaagatcca ttgcatgctt cgacggcttt ggagaatgaa
     2101 accaccatcg aaaccgtcaa cccctccgta agaagtttga aaagcgaacc tgttggtact
     2161 cccaatatag aggaaaacaa agcggactct tccgcagaag ccgtggtaga agaaccgaat
     2221 gaagctgtgg ctgaaagttc tccaaatgaa gaagcaacgg gacagaaaag tgaggatacc
     2281 gatacacatt ctaacgaaca agctgataat ggatttgtac agactgaaga agtagctgaa
     2341 aacaacataa ccacagaaag tgcaggggaa aataacgaac ctgcagatga tgcagcaatg
     2401 gaatttggac gtccagaagc tgaaattgaa actccagaag taaatgagtc tatagaagat
     2461 gccaatgaac ctgcggagga ttccaatgaa cctgtggagg attccaacaa acctgtgaag
     2521 gattccaaca aacctgtgga ggattccaac aaacctgtgg aggattccaa caaacctgtg
     2581 gaggattcca acaaacctgt ggaggatgcc aatgaacctg tggaagatac cagtgaacct
     2641 gtggaggatg ccggtgaacc cgtacaagaa accaacgagt ttactaccga cattgcctct
     2701 ccaagacatc aagaagaaga tatagaactt gaagccgaac ctaaagatgc taccgaaagt
     2761 gttgcagtcg agccatccaa tgaagatgta aaaccagaag aaaaaggttc agaggcagaa
     2821 gacgatatca acaacgtttc caaggaggct gcctctggtg agagtactac ccaccaaaaa
     2881 actgaggcct ctgcttctct tgaaagcagt gccgtcacgg aagaacaaga gacaacggaa
     2941 gccgaagtaa atacagatga cgttttgtcc actaaagaag ctaaaaaaaa tactggcaac
     3001 agcaacagta ataagaagaa gaataagaag aataagaaga aagggaaaaa gaaagcttcg
     3061 tacgctgcag gtcgacggat cggtgacggt gctggtttaa ttaacatgtc taaaggtgaa
     3121 gaattattca ctggtgttgt cccaattttg gttgaattag atggtgatgt taatggtcac
     3181 aaattttctg tctccggtga aggtgaaggt gatgctactt acggtaaatt gaccttaaaa
     3241 tttatttgta ctactggtaa attgccagtt ccatggccaa ccttagtcac tactttaact
     3301 tatggtgttc aatgtttttc tagataccca gatcatatga aacaacatga ctttttcaag
     3361 tctgccatgc cagaaggtta tgttcaagaa agaactattt ttttcaaaga tgacggtaac
     3421 tacaagacca gagctgaagt caagtttgaa ggtgatacct tagttaatag aatcgaatta
     3481 aaaggtattg attttaaaga agatggtaac attttaggtc acaaattgga atacaactat
     3541 aactctcaca atgtttacat catggctgac aaacaaaaga atggtatcaa agttaacttc
     3601 aaaattagac acaacattga agatggttct gttcaattag ctgaccatta tcaacaaaat
     3661 actccaattg gtgatggtcc agtcttgtta ccagacaacc attacttatc cactcaatct
     3721 gccttatcca aagatccaaa cgaaaagaga gaccacatgg tcttgttaga atttgttact
     3781 gctgctggta ttacccatgg tattgatgaa ttgtacaaat aaggcgcgcc acttctaaat
     3841 aagcgaattt cttatgattt atgattttta ttattaaata agttataaaa aaaataagtg
     3901 tatacaaatt ttaaagtgac tcttaggttt taaaacgaaa attcttattc ttgagtaact
     3961 ctttcctgta ggtcaggttg ctttctcagg tatagtatga ggtcgctctt attgaccaca
     4021 cctctaccgg cagatccgct agggataaca gggtaatata gatctgttta gcttgcctcg
     4081 tccccgccgg gtcacccggc cagcgacatg gaggcccaga ataccctcct tgacagtctt
     4141 gacgtgcgca gctcaggggc atgatgtgac tgtcgcccgt acatttagcc catacatccc
     4201 catgtataat catttgcatc catacatttt gatggccgca cggcgcgaag caaaaattac
     4261 ggctcctcgc tgcggacctg cgagcaggga aacgctcccc tcacagacgc gtataacttc
     4321 gtataatgta tgctatacga agttattgaa ttgtccccac gccgcgcccc tgtagagaaa
     4381 tataaaaggt taggatttgc cactgaggtt cttctttcat atacttcctt ttaaaatctt
     4441 gctaggatac agttctcaca tcacatccga acataaacaa ccatgacagt caacactaag
     4501 acctatagtg agagagcaga aactcatgcc tcaccagtag cacaacgatt atttcgatta
     4561 atggaactga agaaaaccaa tttatgtgca tcaattgatg ttgataccac taaggaattc
     4621 cttgaattaa ttgataaatt gggtccttat gtatgcttaa tcaagactca tattgatata
     4681 atcaatgatt tttcctatga atccactatt gaaccattat tagaactttc acgtaaacat
     4741 caatttatga tttttgaaga tagaaaattt gctgatattg gtaataccgt gaagaaacaa
     4801 tatattggtg gagtttataa aattagtagt tgggcagata ttactaatgc tcatggtgtc
     4861 actgggaatg gagtagttga aggattaaaa cagggagcta aagaaaccac caccaaccaa
     4921 gagccaagag ggttattgat gttagctgaa ttatcatcag tgggatcatt agcatatgga
     4981 gaatattctc aaaaaactgt tgaaattgct aaatccgata aggaatttgt tattggattt
     5041 attgcccaac gtgatatggg tggacaagaa gaaggatttg attggcttat tatgacacct
     5101 ggagttggat tagatgataa aggtgatgga ttaggacaac aatatagaac tgttgatgaa
     5161 gttgttagca ctggaactga tattatcatt gttggtagag gattgtttgg taaaggaaga
     5221 gatccagata ttgaaggtaa aaggtataga gatgctggtt ggaatgctta tttgaaaaag
     5281 actggccaat tataatcagt actgacaata aaaagattct tgttttcaag aacttgtcat
     5341 ttgtatagtt tttttatatt gtagttgttc tattttaatc aaatgttagc gtgatttata
     5401 ttttttttcg cctcgacatc atctgcccag atgcgaagtt aagtgcgcag aaagtaatat
     5461 catgcgtcaa tcgtatgtga atgctggtcg ctatactgct gtcgattcga tactaacgcc
     5521 gccatccagt gtcgaaaacg agctcgaatt cataacttcg tataatgtat gctatacgaa
     5581 gttatatcga tgatatcaga tccactagtg gcctatgcgg ccgcggatct gccggtctcc
     5641 ctatagtgag tcgtattaat ttcgataagc caggttaacc tgcattaatg aatcggccaa
     5701 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     5761 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     5821 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     5881 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     5941 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     6001 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     6061 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     6121 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     6181 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     6241 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     6301 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     6361 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     6421 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     6481 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     6541 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     6601 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     6661 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     6721 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     6781 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     6841 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     6901 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     6961 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     7021 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     7081 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     7141 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     7201 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     7261 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     7321 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     7381 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     7441 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     7501 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     7561 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     7621 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc
     7681 attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg
     7741 cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct
     7801 tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc
     7861 gggtgtcggg gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat
     7921 atggacatat tgtcgttaga acgcggctac aattaataca taaccttatg tatcatacac
     7981 atacg
//
