LOCUS       sequence-292116-        7117 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Integration of ERG6-GFP as an additional copy in genome, use 
            auxotrophic marker URA3(Candida albicans). Present on ER and lipid 
            droplets. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7117)
  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 7117)
  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: ZJOM30
FEATURES             Location/Qualifiers
     source          1..7117
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     30..48
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(86..108)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     208..227
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        308..326
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     308..325
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             2545..3261
                     /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      3282..3469
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(3337..3361)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     protein_bind    3752..3785
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     terminator      4740..4937
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     protein_bind    4991..5024
                     /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(5078..5097)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5079..5097)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5184..5201)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5355..5943)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5435..5454)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6114..6905,6906..6974))
                     /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     6737..6756
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6975..7079)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg
       61 cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag
      121 cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg
      181 gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc
      241 atatggacat attgtcgtta gaacgcggct acaattaata cataacctta tgtatcatac
      301 acatacgatt taggtgacac tatagaacgc ggccgccagc tgaatgctcg ctatcctcgc
      361 catcacgtgt accagtacgt ctatgttcat tttctatttc atcgtgacga gattttgaac
      421 cctcaaactc cgctaatgaa cccggagaaa cgacatccag cgacctcttg gcggtagaca
      481 cactcatggt ttaagaaaca acttttcctt ctttaaacgt tttgggtggc aaaaaacttc
      541 ccttaaacaa actaaaaatg gctcgtgttc atgcacaagt caaccagggg ctcaggaaaa
      601 aaagctaaac accaatccta tatataacgt aaaggtactg tgcttaatcg gtaaacggaa
      661 gactattaag caaatgtatc tcttaagacc ttacgcatcc atcagttgcc accctcctac
      721 tcttcatcgg agggtttccg gttcccatga caaagacggc gctttggcgg ggcccaaatt
      781 gttacccgca cccagaaaaa gaaaagagta acctaggtca aatcaaccaa acagctaggg
      841 ctggttacta tttttgaaaa gatgcaacag ggtaagatca gcaaaattta gggccgaaca
      901 aagccccgat cttcgtatat ggtacctcgt tcccgtactt attttcaatt tttggcgtcg
      961 tttactgccg tttccgataa atacaaaaaa gttttcgaga agcgagagta agtgaggagc
     1021 agacaatgta ttattcactg taaactgctc cacttcgtct caatggtaca cttcattttt
     1081 atcgccttac gaagcatgcg ttttatgcga agattggtga gaaacctcca atacttgctg
     1141 ttgccgataa cttcttcatt gctttttata tagttcgggt gttttctcct atcctctgct
     1201 gctctctttt tctttccatt cgtctttgta atttataagc tgttaatttt cattcaagtt
     1261 tcccttatct gttttacttt cgatttaagt tttacataat ttaaaaaaac aagaataaaa
     1321 taataatata gtaggcagca taagatgagt gaaacagaat tgagaaaaag acaggcccaa
     1381 ttcactaggg agttacatgg tgatgatatt ggtaaaaaga caggtttgag tgcattgatg
     1441 tcgaagaaca actctgccca aaaggaagcc gttcagaagt acttgagaaa ttgggatggt
     1501 agaaccgata aagatgccga agaacgtcgt cttgaggatt ataatgaagc cacacattcc
     1561 tactataacg tcgttacaga tttctatgaa tatggttggg gttcctcttt ccatttcagc
     1621 agattttata aaggtgagag tttcgctgcc tcgatagcaa gacatgaaca ttatttagct
     1681 tacaaggctg gtattcaaag aggcgattta gttctcgacg ttggttgtgg tgttgggggc
     1741 ccagcaagag agattgcaag atttaccggt tgtaacgtca tcggtctaaa caataacgat
     1801 taccaaattg ccaaggcaaa atattacgct aaaaaataca atttgagtga ccaaatggac
     1861 tttgtaaagg gtgatttcat gaaaatggat ttcgaagaaa acactttcga caaagtttat
     1921 gcaattgagg ccacatgtca cgctccaaaa ttagaaggtg tatacagcga aatctacaag
     1981 gttttgaaac cgggtggtac ctttgctgtt tacgaatggg taatgactga taaatatgac
     2041 gaaaacaatc ctgaacatag aaagatcgct tatgaaattg aactaggtga tggtatccca
     2101 aagatgttcc atgtcgacgt ggctaggaaa gcattgaaga actgtggttt cgaagtcctc
     2161 gttagcgaag acctggcgga caatgatgat gaaatccctt ggtattaccc attaactggt
     2221 gagtggaagt acgttcaaaa cttagctaat ttggccacat ttttcagaac ttcttacttg
     2281 ggtagacaat ttactacagc aatggttact gtaatggaaa aattaggtct agccccagaa
     2341 ggttccaagg aagttactgc tgctctagaa aatgctgcgg ttggtttagt tgccggtggt
     2401 aagtccaagt tattcactcc aatgatgctt ttcgtcgcta ggaagccaga aaacgccgaa
     2461 accccctccc aaacttccca agaagcaact caagcttcgt acgctgcagg tcgacggatc
     2521 ggtgacggtg ctggtttaat taacatgtct aaaggtgaag aattattcac tggtgttgtc
     2581 ccaattttgg ttgaattaga tggtgatgtt aatggtcaca aattttctgt ctccggtgaa
     2641 ggtgaaggtg atgctactta cggtaaattg accttaaaat ttatttgtac tactggtaaa
     2701 ttgccagttc catggccaac cttagtcact actttaactt atggtgttca atgtttttct
     2761 agatacccag atcatatgaa acaacatgac tttttcaagt ctgccatgcc agaaggttat
     2821 gttcaagaaa gaactatttt tttcaaagat gacggtaact acaagaccag agctgaagtc
     2881 aagtttgaag gtgatacctt agttaataga atcgaattaa aaggtattga ttttaaagaa
     2941 gatggtaaca ttttaggtca caaattggaa tacaactata actctcacaa tgtttacatc
     3001 atggctgaca aacaaaagaa tggtatcaaa gttaacttca aaattagaca caacattgaa
     3061 gatggttctg ttcaattagc tgaccattat caacaaaata ctccaattgg tgatggtcca
     3121 gtcttgttac cagacaacca ttacttatcc actcaatctg ccttatccaa agatccaaac
     3181 gaaaagagag accacatggt cttgttagaa tttgttactg ctgctggtat tacccatggt
     3241 attgatgaat tgtacaaata aggcgcgcca cttctaaata agcgaatttc ttatgattta
     3301 tgatttttat tattaaataa gttataaaaa aaataagtgt atacaaattt taaagtgact
     3361 cttaggtttt aaaacgaaaa ttcttattct tgagtaactc tttcctgtag gtcaggttgc
     3421 tttctcaggt atagtatgag gtcgctctta ttgaccacac ctctaccggc agatccgcta
     3481 gggataacag ggtaatatag atctgtttag cttgcctcgt ccccgccggg tcacccggcc
     3541 agcgacatgg aggcccagaa taccctcctt gacagtcttg acgtgcgcag ctcaggggca
     3601 tgatgtgact gtcgcccgta catttagccc atacatcccc atgtataatc atttgcatcc
     3661 atacattttg atggccgcac ggcgcgaagc aaaaattacg gctcctcgct gcggacctgc
     3721 gagcagggaa acgctcccct cacagacgcg tataacttcg tataatgtat gctatacgaa
     3781 gttattgaat tgtccccacg ccgcgcccct gtagagaaat ataaaaggtt aggatttgcc
     3841 actgaggttc ttctttcata tacttccttt taaaatcttg ctaggataca gttctcacat
     3901 cacatccgaa cataaacaac catgacagtc aacactaaga cctatagtga gagagcagaa
     3961 actcatgcct caccagtagc acaacgatta tttcgattaa tggaactgaa gaaaaccaat
     4021 ttatgtgcat caattgatgt tgataccact aaggaattcc ttgaattaat tgataaattg
     4081 ggtccttatg tatgcttaat caagactcat attgatataa tcaatgattt ttcctatgaa
     4141 tccactattg aaccattatt agaactttca cgtaaacatc aatttatgat ttttgaagat
     4201 agaaaatttg ctgatattgg taataccgtg aagaaacaat atattggtgg agtttataaa
     4261 attagtagtt gggcagatat tactaatgct catggtgtca ctgggaatgg agtagttgaa
     4321 ggattaaaac agggagctaa agaaaccacc accaaccaag agccaagagg gttattgatg
     4381 ttagctgaat tatcatcagt gggatcatta gcatatggag aatattctca aaaaactgtt
     4441 gaaattgcta aatccgataa ggaatttgtt attggattta ttgcccaacg tgatatgggt
     4501 ggacaagaag aaggatttga ttggcttatt atgacacctg gagttggatt agatgataaa
     4561 ggtgatggat taggacaaca atatagaact gttgatgaag ttgttagcac tggaactgat
     4621 attatcattg ttggtagagg attgtttggt aaaggaagag atccagatat tgaaggtaaa
     4681 aggtatagag atgctggttg gaatgcttat ttgaaaaaga ctggccaatt ataatcagta
     4741 ctgacaataa aaagattctt gttttcaaga acttgtcatt tgtatagttt ttttatattg
     4801 tagttgttct attttaatca aatgttagcg tgatttatat tttttttcgc ctcgacatca
     4861 tctgcccaga tgcgaagtta agtgcgcaga aagtaatatc atgcgtcaat cgtatgtgaa
     4921 tgctggtcgc tatactgctg tcgattcgat actaacgccg ccatccagtg tcgaaaacga
     4981 gctcgaattc ataacttcgt ataatgtatg ctatacgaag ttatatcgat gatatcagat
     5041 ccactagtgg cctatgcggc cgcggatctg ccggtctccc tatagtgagt cgtattaatt
     5101 tcgataagcc aggttaacct gcattaatga atcggccaac gcgcggggag aggcggtttg
     5161 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     5221 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     5281 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     5341 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     5401 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     5461 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     5521 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     5581 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     5641 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     5701 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     5761 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     5821 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     5881 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     5941 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     6001 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     6061 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     6121 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     6181 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     6241 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     6301 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     6361 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     6421 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     6481 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     6541 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     6601 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     6661 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     6721 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     6781 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     6841 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     6901 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     6961 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     7021 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     7081 acatttcccc gaaaagtgcc acctgacgtc taagaaa
//
