LOCUS       sequence-267443-        6983 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Integration of VRG4-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 6983)
  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 6983)
  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: ZJOM2
FEATURES             Location/Qualifiers
     source          1..6983
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             2085..2801
                     /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      2822..3009
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2877..2901)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     protein_bind    3292..3325
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     terminator      4280..4477
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     protein_bind    4531..4564
                     /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(4618..4637)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4619..4637)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4724..4741)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4895..5483)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4975..4994)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5654..6445,6446..6514))
                     /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     6277..6296
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6515..6619)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6687..6705
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6743..6765)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6865..6884
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        6965..6983
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     6965..6982
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 acgcggccgc cagctgaagc ttgatctcaa ggaaaaaaat gcaatgtcgg gcgccaatac
       61 ttatgatcat gcttagtctc gacattgacc ttgcttgcta cagtaatctg gtgtccgagt
      121 agtgacatct aggtaccggt gagaactata tcttcattac tacacttgtc gcccctgaat
      181 attatcgcta cacacgcgaa agagctcttg ttaaagttac tctcgcatca ttgcggtaca
      241 tgttttgcat attcatctga agtaggaaag ggtataagca gtatatgatg taccatctgc
      301 ataagcggtg tttaatgcgt acctatcgtt taagaacaaa aggaacacac ccacacccgc
      361 tccaatacca tattgagaga ataatgaggt aacatatgct tctatttggt tcgataagac
      421 ggctactagt ccaaaaaaag tcaaacaaca atttcaacag ctacattctg aagatagaac
      481 gcatttttgc ctgaatattg caactcaata gacattctaa agtgctacga gaaaaaagct
      541 tgtaatggac catggcaatg tgcgccatat atatatgcgg ctcccttatg agtagaagaa
      601 aatatatgac tggtggtaca aagtggcatg gtggtatata gatgtaggaa caccatttga
      661 gtttgccata atgattaaat ttcccttttt ataaagaatt agccgcccat aacacgaaaa
      721 atatcacagt tgacgaaaga agacacgtcg cctaaacgcg taatgaatcg caatatacga
      781 aaagctaaac gggtttttca gaatcctctt acccagcctt ggataatagt tttggccgaa
      841 aaaaaaaaaa agaaagaaaa aaattgaaat ttcgcgatcc gaacaaacaa tgaacagaaa
      901 aacagaatat atataacgct gccaagccta tttcttgttg ccgattaatt gttgacatcc
      961 tcttaattac caaaagagcc taagaaaaca aacacactaa ccacacagta cctttcgccc
     1021 gaatgtctga attgaaaaca ggtcatgcag gccataaccc ttgggcttca gttgccaatt
     1081 ccggtccgat ctctatttta tcctactgtg gttcctctat tttaatgacg gtgactaaca
     1141 agttcgtcgt caatttgaag gatttcaaca tgaactttgt catgcttttc gtgcaatctt
     1201 tggtttgtac tataaccttg attatcctac gtatactggg ctatgcgaag ttccgttcat
     1261 taaacaaaac agacgccaag aactggttcc ctatttcctt tttactggtc ttgatgatct
     1321 acacctcttc gaaggcttta caatacttgg ctgttccaat ttacaccatt ttcaagaatt
     1381 tgactattat cttgattgct tatggtgagg ttctcttttt tggtggctct gtcacctcca
     1441 tggaattgtc atcatttttg ttgatggtcc tttcttctgt cgttgcaact tggggtgacc
     1501 agcaagctgt ggctgccaag gctgcttcat tggctgaagg agcagccggt gctgttgcct
     1561 cctttaaccc aggttatttc tggatgttca ccaactgtat cacttctgca ttattcgttc
     1621 ttataatgag aaagagaatt aagttaacta acttcaagga tttcgacact atgttttaca
     1681 acaatgtttt ggctctacct attctattgc tgttttcttt ctgtgtggaa gattggtctt
     1741 cagttaattt gaccaataac ttttctaacg attcgctaac tgctatgatc atcagtggtg
     1801 ttgcatccgt cggtatttct tactgttccg gttggtgtgt tcgtgttact tcgtctacta
     1861 catattcgat ggtaggggct ttgaacaagc tgccaattgc cttgtctggt ttgattttct
     1921 ttgatgctcc aagaaacttc ttatctattc tctccatttt tattggtttc ctatcaggta
     1981 ttatttatgc tgttgccaaa caaaagaagc aacaagccca acctttacgt aaagcttcgt
     2041 acgctgcagg tcgacggatc ggtgacggtg ctggtttaat taacatgtct aaaggtgaag
     2101 aattattcac tggtgttgtc ccaattttgg ttgaattaga tggtgatgtt aatggtcaca
     2161 aattttctgt ctccggtgaa ggtgaaggtg atgctactta cggtaaattg accttaaaat
     2221 ttatttgtac tactggtaaa ttgccagttc catggccaac cttagtcact actttaactt
     2281 atggtgttca atgtttttct agatacccag atcatatgaa acaacatgac tttttcaagt
     2341 ctgccatgcc agaaggttat gttcaagaaa gaactatttt tttcaaagat gacggtaact
     2401 acaagaccag agctgaagtc aagtttgaag gtgatacctt agttaataga atcgaattaa
     2461 aaggtattga ttttaaagaa gatggtaaca ttttaggtca caaattggaa tacaactata
     2521 actctcacaa tgtttacatc atggctgaca aacaaaagaa tggtatcaaa gttaacttca
     2581 aaattagaca caacattgaa gatggttctg ttcaattagc tgaccattat caacaaaata
     2641 ctccaattgg tgatggtcca gtcttgttac cagacaacca ttacttatcc actcaatctg
     2701 ccttatccaa agatccaaac gaaaagagag accacatggt cttgttagaa tttgttactg
     2761 ctgctggtat tacccatggt attgatgaat tgtacaaata aggcgcgcca cttctaaata
     2821 agcgaatttc ttatgattta tgatttttat tattaaataa gttataaaaa aaataagtgt
     2881 atacaaattt taaagtgact cttaggtttt aaaacgaaaa ttcttattct tgagtaactc
     2941 tttcctgtag gtcaggttgc tttctcaggt atagtatgag gtcgctctta ttgaccacac
     3001 ctctaccggc agatccgcta gggataacag ggtaatatag atctgtttag cttgcctcgt
     3061 ccccgccggg tcacccggcc agcgacatgg aggcccagaa taccctcctt gacagtcttg
     3121 acgtgcgcag ctcaggggca tgatgtgact gtcgcccgta catttagccc atacatcccc
     3181 atgtataatc atttgcatcc atacattttg atggccgcac ggcgcgaagc aaaaattacg
     3241 gctcctcgct gcggacctgc gagcagggaa acgctcccct cacagacgcg tataacttcg
     3301 tataatgtat gctatacgaa gttattgaat tgtccccacg ccgcgcccct gtagagaaat
     3361 ataaaaggtt aggatttgcc actgaggttc ttctttcata tacttccttt taaaatcttg
     3421 ctaggataca gttctcacat cacatccgaa cataaacaac catgacagtc aacactaaga
     3481 cctatagtga gagagcagaa actcatgcct caccagtagc acaacgatta tttcgattaa
     3541 tggaactgaa gaaaaccaat ttatgtgcat caattgatgt tgataccact aaggaattcc
     3601 ttgaattaat tgataaattg ggtccttatg tatgcttaat caagactcat attgatataa
     3661 tcaatgattt ttcctatgaa tccactattg aaccattatt agaactttca cgtaaacatc
     3721 aatttatgat ttttgaagat agaaaatttg ctgatattgg taataccgtg aagaaacaat
     3781 atattggtgg agtttataaa attagtagtt gggcagatat tactaatgct catggtgtca
     3841 ctgggaatgg agtagttgaa ggattaaaac agggagctaa agaaaccacc accaaccaag
     3901 agccaagagg gttattgatg ttagctgaat tatcatcagt gggatcatta gcatatggag
     3961 aatattctca aaaaactgtt gaaattgcta aatccgataa ggaatttgtt attggattta
     4021 ttgcccaacg tgatatgggt ggacaagaag aaggatttga ttggcttatt atgacacctg
     4081 gagttggatt agatgataaa ggtgatggat taggacaaca atatagaact gttgatgaag
     4141 ttgttagcac tggaactgat attatcattg ttggtagagg attgtttggt aaaggaagag
     4201 atccagatat tgaaggtaaa aggtatagag atgctggttg gaatgcttat ttgaaaaaga
     4261 ctggccaatt ataatcagta ctgacaataa aaagattctt gttttcaaga acttgtcatt
     4321 tgtatagttt ttttatattg tagttgttct attttaatca aatgttagcg tgatttatat
     4381 tttttttcgc ctcgacatca tctgcccaga tgcgaagtta agtgcgcaga aagtaatatc
     4441 atgcgtcaat cgtatgtgaa tgctggtcgc tatactgctg tcgattcgat actaacgccg
     4501 ccatccagtg tcgaaaacga gctcgaattc ataacttcgt ataatgtatg ctatacgaag
     4561 ttatatcgat gatatcagat ccactagtgg cctatgcggc cgcggatctg ccggtctccc
     4621 tatagtgagt cgtattaatt tcgataagcc aggttaacct gcattaatga atcggccaac
     4681 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     4741 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     4801 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     4861 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     4921 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     4981 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     5041 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     5101 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     5161 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     5221 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     5281 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     5341 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     5401 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     5461 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     5521 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     5581 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     5641 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     5701 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     5761 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     5821 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     5881 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     5941 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     6001 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     6061 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     6121 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     6181 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     6241 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     6301 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     6361 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     6421 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     6481 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     6541 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     6601 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca
     6661 ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc
     6721 gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt
     6781 gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg
     6841 ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata
     6901 tggacatatt gtcgttagaa cgcggctaca attaatacat aaccttatgt atcatacaca
     6961 tacgatttag gtgacactat aga
//
