LOCUS       sequence-265887-        6139 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Integration of GFP-TLG1 as an additional copy in genome, use 
            auxotrophic marker URA3(Saccharomyces cerevisiae). Marker for yeast 
            late golgi/early endosome..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6139)
  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 6139)
  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: ZJOM11
FEATURES             Location/Qualifiers
     source          1..6139
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     112..129
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    246..267
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(282..287,288..305,306..312)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    320..336
                     /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     325..347
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     344..360
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     344..360
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     379..399
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        381..399
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        418..652
                     /gene="S. cerevisiae CUP1"
                     /label=CUP1 promoter
                     /note="inducible metallothionein promoter, regulated by 
                     Cu2+"
     CDS             663..1376
                     /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(2293..2312)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2294..2312)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2322..2339)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2322..2338)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2331..2353)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2479..2934
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2566..2585)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2776..2797
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(3065..3868)
                     /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(3869..4084)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(4108..4127)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4227..4249
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4287..4305)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4373..4477
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4478..4546,4547..5338)
                     /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(4696..4715)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5509..6097
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5998..6017
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
       61 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
      121 gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg
      181 attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac
      241 gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg
      301 gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac
      361 catgattacg ccaagcgcgc aattaaccct cactaaaggg aacaaaagct ggagctccta
      421 gttagaaaaa gacatttttg ctgtcagtca ctgtcaagag attcttttgc tggcatttct
      481 tctagaagca aaaagagcga tgcgtctttt ccgctgaacc gttccagcaa aaaagactac
      541 caacgcaata tggattgtca gaatcatata aaagagaagc aaataactcc ttgtcttgta
      601 tcaattgcat tataatatct tcttgttagt gcaatatcat atagaagtca tcgactagta
      661 acatgtctaa aggtgaagaa ttattcactg gtgttgtccc aattttggtt gaattagatg
      721 gtgatgttaa tggtcacaaa ttttctgtct ccggtgaagg tgaaggtgat gctacttacg
      781 gtaaattgac cttaaaattt atttgtacta ctggtaaatt gccagttcca tggccaacct
      841 tagtcactac tttaacttat ggtgttcaat gtttttctag atacccagat catatgaaac
      901 aacatgactt tttcaagtct gccatgccag aaggttatgt tcaagaaaga actatttttt
      961 tcaaagatga cggtaactac aagaccagag ctgaagtcaa gtttgaaggt gataccttag
     1021 ttaatagaat cgaattaaaa ggtattgatt ttaaagaaga tggtaacatt ttaggtcaca
     1081 aattggaata caactataac tctcacaatg tttacatcat ggctgacaaa caaaagaatg
     1141 gtatcaaagt taacttcaaa attagacaca acattgaaga tggttctgtt caattagctg
     1201 accattatca acaaaatact ccaattggtg atggtccagt cttgttacca gacaaccatt
     1261 acttatccac tcaatctgcc ttatccaaag atccaaacga aaagagagac cacatggtct
     1321 tgttagaatt tgttactgct gctggtatta cccatggtat tgatgaattg tacaaaggag
     1381 ccggaacaag tgacatgaag atgaacaaca gtgaagatcc gtttcaacaa gttgttaagg
     1441 acaccaagga gcaattgaac cgcataaaca attacataac tcgtcacaat actgctggtg
     1501 atgacgatca agaggaggaa atacaagata ttttaaagga tgttgaggaa acaatagttg
     1561 atttggacag aagcataatc gtaatgaaaa gggatgaaaa cgaagacgtg agtggtaggg
     1621 aagcacaagt taaaaatata aaacagcaac ttgatgctct gaagttgcgt tttgatcgaa
     1681 gaatacagga atctactcaa acaactattc ctctagaaga gacggtggaa aattcaacac
     1741 ttaacaccag catggctgag aacaatgatg gtggtatgtc caatccgttt caggaacaaa
     1801 tgttaagaga acaagatgtt catttagatg gtattcacaa gacaatgcaa aatttgcata
     1861 ttcaagctca aacaatgggg gatgaattag agaaccaggg acaattgttg gataatatgg
     1921 acgagggtat ggacggtgtt gtaaataagc tggctagagg tcgtaggcaa ttggaatggg
     1981 tctacgaaaa aaataaagaa aaatacgacg attgttgtat aggacttctt attgtcgtct
     2041 tgatagtttt attagttttg gcattcattg cttagtcttt tatatgaaaa gaaatgaagc
     2101 gttaacatct aaaatcgaaa ttatagggac tcattagcca tctattgttc cagtataaga
     2161 atgtttacat aaaaaattcg aaaaaaaaag ataaaataag ggaaaaatat atatatattc
     2221 attaagaagt tttgtacaca acggagccaa gatttcacca gtatctcgag ggggggcccg
     2281 gtacccaatt cgccctatag tgagtcgtat tacgcgcgct cactggccgt cgttttacaa
     2341 cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct
     2401 ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc
     2461 agcctgaatg gcgaatggac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     2521 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     2581 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     2641 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
     2701 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     2761 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     2821 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
     2881 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttcctgat
     2941 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatagggt aataactgat
     3001 ataattaaat tgaagctcta atttgtgagt ttagtataca tgcatttact tataatacag
     3061 ttttttagtt ttgctggccg catcttctca aatatgcttc ccagcctgct tttctgtaac
     3121 gttcaccctc taccttagca tcccttccct ttgcaaatag tcctcttcca acaataataa
     3181 tgtcagatcc tgtagagacc acatcatcca cggttctata ctgttgaccc aatgcgtctc
     3241 ccttgtcatc taaacccaca ccgggtgtca taatcaacca atcgtaacct tcatctcttc
     3301 cacccatgtc tctttgagca ataaagccga taacaaaatc tttgtcgctc ttcgcaatgt
     3361 caacagtacc cttagtatat tctccagtag atagggagcc cttgcatgac aattctgcta
     3421 acatcaaaag gcctctaggt tcctttgtta cttcttctgc cgcctgcttc aaaccgctaa
     3481 caatacctgg gcccaccaca ccgtgtgcat tcgtaatgtc tgcccattct gctattctgt
     3541 atacacccgc agagtactgc aatttgactg tattaccaat gtcagcaaat tttctgtctt
     3601 cgaagagtaa aaaattgtac ttggcggata atgcctttag cggcttaact gtgccctcca
     3661 tggaaaaatc agtcaagata tccacatgtg tttttagtaa acaaattttg ggacctaatg
     3721 cttcaactaa ctccagtaat tccttggtgg tacgaacatc caatgaagca cacaagtttg
     3781 tttgcttttc gtgcatgata ttaaatagct tggcagcaac aggactagga tgagtagcag
     3841 cacgttcctt atatgtagct ttcgacatga tttatcttcg tttcctgcag gtttttgttc
     3901 tgtgcagttg ggttaagaat actgggcaat ttcatgtttc ttcaacacta catatgcgta
     3961 tatataccaa tctaagtctg tgctccttcc ttcgttcttc cttctgttcg gagattaccg
     4021 aatcaaaaaa atttcaagga aaccgaaatc aaaaaaaaga ataaaaaaaa aatgatgaat
     4081 tgaaaaggtg gtatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     4141 gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc
     4201 cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc
     4261 atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt
     4321 catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac
     4381 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     4441 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     4501 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     4561 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     4621 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     4681 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     4741 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     4801 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     4861 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     4921 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     4981 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     5041 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     5101 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     5161 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     5221 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     5281 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     5341 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     5401 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     5461 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     5521 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     5581 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     5641 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     5701 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     5761 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     5821 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     5881 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     5941 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     6001 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     6061 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     6121 acggttcctg gccttttgc
//
