LOCUS       sequence-268964-        7302 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Integration of GFP-PHO8 as an additional copy in genome, use 
            auxotrophic marker URA3(Saccharomyces cerevisiae). Marker for yeast 
            vacuole..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7302)
  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 7302)
  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: ZJOM27
FEATURES             Location/Qualifiers
     source          1..7302
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..235
                     /gene="S. cerevisiae CUP1"
                     /label=CUP1 promoter
                     /note="inducible metallothionein promoter, regulated by 
                     Cu2+"
     CDS             246..959
                     /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"
     primer_bind     complement(3039..3058)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3040..3058)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3068..3085)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3068..3084)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3077..3099)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3225..3680
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3312..3331)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3522..3543
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(3811..4614)
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     promoter        complement(4615..4830)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(4854..4873)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4973..4995
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5033..5051)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5119..5223
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5224..5292,5293..6084)
                     /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     complement(5442..5461)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6255..6843
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6744..6763
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6997..7014
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7131..7152
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7167..7172,7173..7190,7191..7197)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7205..7221
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     7210..7232
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7229..7245
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7229..7245
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     7264..7284
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        7266..7284
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 ctagttagaa aaagacattt ttgctgtcag tcactgtcaa gagattcttt tgctggcatt
       61 tcttctagaa gcaaaaagag cgatgcgtct tttccgctga accgttccag caaaaaagac
      121 taccaacgca atatggattg tcagaatcat ataaaagaga agcaaataac tccttgtctt
      181 gtatcaattg cattataata tcttcttgtt agtgcaatat catatagaag tcatcgacta
      241 gtaacatgtc taaaggtgaa gaattattca ctggtgttgt cccaattttg gttgaattag
      301 atggtgatgt taatggtcac aaattttctg tctccggtga aggtgaaggt gatgctactt
      361 acggtaaatt gaccttaaaa tttatttgta ctactggtaa attgccagtt ccatggccaa
      421 ccttagtcac tactttaact tatggtgttc aatgtttttc tagataccca gatcatatga
      481 aacaacatga ctttttcaag tctgccatgc cagaaggtta tgttcaagaa agaactattt
      541 ttttcaaaga tgacggtaac tacaagacca gagctgaagt caagtttgaa ggtgatacct
      601 tagttaatag aatcgaatta aaaggtattg attttaaaga agatggtaac attttaggtc
      661 acaaattgga atacaactat aactctcaca atgtttacat catggctgac aaacaaaaga
      721 atggtatcaa agttaacttc aaaattagac acaacattga agatggttct gttcaattag
      781 ctgaccatta tcaacaaaat actccaattg gtgatggtcc agtcttgtta ccagacaacc
      841 attacttatc cactcaatct gccttatcca aagatccaaa cgaaaagaga gaccacatgg
      901 tcttgttaga atttgttact gctgctggta ttacccatgg tattgatgaa ttgtacaaag
      961 gagccggaac aagtgacatg atgactcaca cattaccaag cgaacagaca cgtcttgttc
     1021 ctggatctga ctcgagctct cgtcctaaaa agagacggat ctcgaagaga tcaaagataa
     1081 tagtatccac tgtggtctgt attggtttgt tgctggtttt agtgcaactg gcatttccaa
     1141 gcagttttgc attacgttct gcatcacaca agaagaagaa tgtcatattc ttcgtgacgg
     1201 atgggatggg acctgcgtct ttgtccatgg caagatcatt caaccagcat gttaatgatt
     1261 tacctataga cgacatcctg acacttgacg agcattttat cgggtcttcg agaacaaggt
     1321 cctcggactc actagtcact gactcagctg cgggcgccac cgcatttgca tgtgcattaa
     1381 agagctataa tggggccatc ggtgttgatc cgcatcacag accttgtggt acagtgctcg
     1441 aggcggctaa gttagcaggt tatctcactg ggcttgttgt tacgacaaga atcacagatg
     1501 ccactccggc ttcattcagt tcacatgttg attacagatg gcaggaagat cttatcgcta
     1561 cgcaccagtt aggcgaatat cctctgggga gagtagtaga cttactcatg ggtggcggca
     1621 ggagccattt ctaccctcaa ggcgaaaagg cttctccata tggtcaccac ggtgctagaa
     1681 aagatggaag agatttaatc gatgaggctc aatctaatgg ctggcagtac gtgggtgacc
     1741 gtaaaaattt tgattctttg ttgaaaagcc acggtgaaaa cgttacttta cctttcttgg
     1801 gtctctttgc agataacgac attccttttg aaattgacag agatgaaaag gagtatcctt
     1861 cccttaagga acaagtcaag gtggctttgg gtgccttgga aaaagcttcc aatgaggaca
     1921 aagattctaa tgggtttttc ttaatggtgg aaggctccag aattgatcat gctggtcacc
     1981 aaaacgatcc agcttcgcag gtaagggagg tgttggcttt cgatgaagca ttccaatacg
     2041 ttttggagtt tgctgaaaat tccgatacgg agactgtttt ggtctctacg tcggatcatg
     2101 aaacgggtgg tttagtcact tcaagacaag tcactgctag ctatcctcag tatgtctggt
     2161 atccacaggt gctcgccaac gccacccatt caggtgagtt tttgaaaaga aaactagtag
     2221 attttgttca tgagcataag ggcgcaagca gcaaaataga aaatttcatc aaacatgaga
     2281 tcctggaaaa agatttgggt atttacgatt acacggatag cgatttagaa acattaattc
     2341 atttggatga caacgcaaat gctatccaag acaaactgaa tgacatggta tcatttaggg
     2401 cccaaattgg ctggaccacg catggtcaca gcgcagtcga cgtcaacata tatgcatacg
     2461 ccaataagaa agctacatgg tcatatgtct tgaacaattt acaaggcaat cacgaaaata
     2521 cggaagttgg ccagttctta gaaaactttt tagaattgaa cctgaacgaa gtcactgatt
     2581 tgattaggga caccaaacac acatcagatt ttgatgctac agaaatagct agtgaagtac
     2641 aacattatga cgaatactac catgagttga ccaactgatc ctatctaact ctccctcttt
     2701 tttcacatat tatttaatac gttataattt atttgaatat atatatatat atatatgtat
     2761 atatactaac cgcatgacac gtactgtatc gcatttcgtt tcggagtgca ggcgagtttt
     2821 aggtgcattc tatagcgtga agaagtgaaa tctcgtcatt tcttcaagtt atgccattca
     2881 gtccagattg tttcacatgc tttcatatag tcatttttgt cgtcgcgaac ttcaaatttg
     2941 aaaacatcat catacagaga gatttctcca ataatgatcc tttctttgtc gttgtcatca
     3001 tcggcatcgt cgttaagtga agcgcggtac ccaattcgcc ctatagtgag tcgtattacg
     3061 cgcgctcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac
     3121 ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca
     3181 ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atggacgcgc cctgtagcgg
     3241 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     3301 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     3361 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     3421 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     3481 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     3541 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
     3601 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa
     3661 aatattaacg cttacaattt cctgatgcgg tattttctcc ttacgcatct gtgcggtatt
     3721 tcacaccgca tagggtaata actgatataa ttaaattgaa gctctaattt gtgagtttag
     3781 tatacatgca tttacttata atacagtttt ttagttttgc tggccgcatc ttctcaaata
     3841 tgcttcccag cctgcttttc tgtaacgttc accctctacc ttagcatccc ttccctttgc
     3901 aaatagtcct cttccaacaa taataatgtc agatcctgta gagaccacat catccacggt
     3961 tctatactgt tgacccaatg cgtctccctt gtcatctaaa cccacaccgg gtgtcataat
     4021 caaccaatcg taaccttcat ctcttccacc catgtctctt tgagcaataa agccgataac
     4081 aaaatctttg tcgctcttcg caatgtcaac agtaccctta gtatattctc cagtagatag
     4141 ggagcccttg catgacaatt ctgctaacat caaaaggcct ctaggttcct ttgttacttc
     4201 ttctgccgcc tgcttcaaac cgctaacaat acctgggccc accacaccgt gtgcattcgt
     4261 aatgtctgcc cattctgcta ttctgtatac acccgcagag tactgcaatt tgactgtatt
     4321 accaatgtca gcaaattttc tgtcttcgaa gagtaaaaaa ttgtacttgg cggataatgc
     4381 ctttagcggc ttaactgtgc cctccatgga aaaatcagtc aagatatcca catgtgtttt
     4441 tagtaaacaa attttgggac ctaatgcttc aactaactcc agtaattcct tggtggtacg
     4501 aacatccaat gaagcacaca agtttgtttg cttttcgtgc atgatattaa atagcttggc
     4561 agcaacagga ctaggatgag tagcagcacg ttccttatat gtagctttcg acatgattta
     4621 tcttcgtttc ctgcaggttt ttgttctgtg cagttgggtt aagaatactg ggcaatttca
     4681 tgtttcttca acactacata tgcgtatata taccaatcta agtctgtgct ccttccttcg
     4741 ttcttccttc tgttcggaga ttaccgaatc aaaaaaattt caaggaaacc gaaatcaaaa
     4801 aaaagaataa aaaaaaaatg atgaattgaa aaggtggtat ggtgcactct cagtacaatc
     4861 tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc
     4921 tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc
     4981 tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg
     5041 atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc
     5101 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     5161 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     5221 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     5281 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     5341 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     5401 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     5461 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     5521 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     5581 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     5641 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     5701 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     5761 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     5821 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     5881 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     5941 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     6001 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     6061 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     6121 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     6181 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     6241 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     6301 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     6361 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     6421 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     6481 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     6541 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     6601 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     6661 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     6721 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     6781 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     6841 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     6901 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     6961 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     7021 gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc
     7081 tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt
     7141 tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt
     7201 ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag
     7261 cgcgcaatta accctcacta aagggaacaa aagctggagc tc
//
