LOCUS       sequence-269549-        6658 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Integration of GFP-TLG2 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 6658)
  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 6658)
  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: ZJOM12
FEATURES             Location/Qualifiers
     source          1..6658
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     133..152
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     386..403
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    520..541
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(556..561,562..579,580..586)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    594..610
                     /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     599..621
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     618..634
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     618..634
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     653..673
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        655..673
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        692..926
                     /gene="S. cerevisiae CUP1"
                     /label=CUP1 promoter
                     /note="inducible metallothionein promoter, regulated by 
                     Cu2+"
     CDS             937..1650
                     /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(3086..3105)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3087..3105)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3115..3132)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3115..3131)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3124..3146)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3272..3727
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3359..3378)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3569..3590
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(3858..4661)
                     /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(4662..4877)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(4901..4920)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5020..5042
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5080..5098)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5166..5270
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5271..5339,5340..6131)
                     /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(5489..5508)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(6302..6658,1..232)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
       61 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
      121 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
      181 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
      241 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
      301 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
      361 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc
      421 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag
      481 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca
      541 ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag
      601 cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc gcgcaattaa
      661 ccctcactaa agggaacaaa agctggagct cctagttaga aaaagaaatt tttgctgtca
      721 gtcactgtca agagattctt ttgctggcat ttcttctaga agcaaaaaga gcgatgcgtc
      781 ttttccgctg aaccgttcca gcaaaaaaga ctaccaacgc aatatggatt gtcagaatca
      841 tataaaagag aagcaaataa ctccttgtct tgtatcaatt gcattataat atcttcttgt
      901 tagtgcaata tcatatagaa gtcatcgact agtaacatgt ctaaaggtga agaattattc
      961 actggtgttg tcccaatttt ggttgaatta gatggtgatg ttaatggtca caaattttct
     1021 gtctccggtg aaggtgaagg tgatgctact tacggtaaat tgaccttaaa atttatttgt
     1081 actactggta aattgccagt tccatggcca accttagtca ctactttaac ttatggtgtt
     1141 caatgttttt ctagataccc agatcatatg aaacaacatg actttttcaa gtctgccatg
     1201 ccagaaggtt atgttcaaga aagaactatt tttttcaaag atgacggtaa ctacaagacc
     1261 agagctgaag tcaagtttga aggtgatacc ttagttaata gaatcgaatt aaaaggtatt
     1321 gattttaaag aagatggtaa cattttaggt cacaaattgg aatacaacta taactctcac
     1381 aatgtttaca tcatggctga caaacaaaag aatggtatca aagttaactt caaaattaga
     1441 cacaacattg aagatggttc tgttcaatta gctgaccatt atcaacaaaa tactccaatt
     1501 ggtgatggtc cagtcttgtt accagacaac cattacttat ccactcaatc tgccttatcc
     1561 aaagatccaa acgaaaagag agaccacatg gtcttgttag aatttgttac tgctgctggt
     1621 attacccatg gtattgatga attgtacaaa ggagccggaa caagtgacat gaagatgttt
     1681 agagatagaa ctaatttatt tttatcatac cgtaggactt tccctcataa cataactttc
     1741 tcatctggga aagcaccctt gggcgatgat caagatattg aaatgggtac atacccgatg
     1801 atgaatatgt ctcacgatat atcggccaga ttgacagatg aacgaaaaaa caagcatgag
     1861 aatcacagtg atgcgttacc acccatattt attgatatag cgcaggatgt tgatgattat
     1921 cttctggagg tgagacggtt aagtgagcag ttggcgaagg tttataggaa gaattcgtta
     1981 cctggctttg aagacaagag tcatgatgag gcgctgattg aagatctaag tttcaaagta
     2041 attcagatgc tgcagaaatg ttacgctgtt atgaaacgat taaagaccat atacaactct
     2101 caatttgtgg atggtaagca attaagtcga gaagagctga ttatactaga taatctacag
     2161 aagatctatg cagaaaagat acaaactgaa agtaataagt tcagagtatt acaaaataat
     2221 tacttgaaat tcttgaacaa ggatgatttg aaacctatcc gcaataaggc cagtgcagag
     2281 aacacgctat tattagatga tgaggaagag gaggcagcaa gagagaagag agagggcctc
     2341 gatattgaag actattcaaa aaggacgttg cagagacagc aacagcttca tgacacttct
     2401 gcagaagcat atttgagaga acgtgacgaa gaaattactc agttggcaag gggtgtatta
     2461 gaagtgagca ctattttcag ggaaatgcaa gatttggttg ttgaccaggg gactattgtg
     2521 gacaggattg attataatct ggaaaataca gtggtagagc taaaaagtgc agataaagag
     2581 ttgaataaag caacacacta ccagaagaga actcaaaaat gtaaagtcat tttattacta
     2641 acattgtgcg taatagcgct ctttttcttt gttatgttga aaccacatgg cggtggcagt
     2701 ggtggtcgta ataatggtag caacaagtac aacaatgatg ataataaaac agttaataac
     2761 agtcatgatg acggcagtaa tacccatatt aatgacgaag agagcaactt gccgagtatc
     2821 gttgaagtaa cagaatctga gaatgatgct ttggatgacc tactttagtc ttttatatga
     2881 aaagaaatga agcgttaaca tctaaaatcg aaattatagg gactcattag ccatctattg
     2941 ttccagtata agaatgttta cataaaaaat tcgaaaaaaa aagataaaat aagggaaaaa
     3001 tatatatata ttcattaaga agttttgtac acaacggagc caagatttca ccagtatctc
     3061 gagggggggc ccggtaccca attcgcccta tagtgagtcg tattacgcgc gctcactggc
     3121 cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc
     3181 agcacatccc cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc
     3241 ccaacagttg cgcagcctga atggcgaatg gacgcgccct gtagcggcgc attaagcgcg
     3301 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     3361 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     3421 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     3481 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     3541 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     3601 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     3661 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt
     3721 acaatttcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatag
     3781 ggtaataact gatataatta aattgaagct ctaatttgtg agtttagtat acatgcattt
     3841 acttataata cagtttttta gttttgctgg ccgcatcttc tcaaatatgc ttcccagcct
     3901 gcttttctgt aacgttcacc ctctacctta gcatcccttc cctttgcaaa tagtcctctt
     3961 ccaacaataa taatgtcaga tcctgtagag accacatcat ccacggttct atactgttga
     4021 cccaatgcgt ctcccttgtc atctaaaccc acaccgggtg tcataatcaa ccaatcgtaa
     4081 ccttcatctc ttccacccat gtctctttga gcaataaagc cgataacaaa atctttgtcg
     4141 ctcttcgcaa tgtcaacagt acccttagta tattctccag tagataggga gcccttgcat
     4201 gacaattctg ctaacatcaa aaggcctcta ggttcctttg ttacttcttc tgccgcctgc
     4261 ttcaaaccgc taacaatacc tgggcccacc acaccgtgtg cattcgtaat gtctgcccat
     4321 tctgctattc tgtatacacc cgcagagtac tgcaatttga ctgtattacc aatgtcagca
     4381 aattttctgt cttcgaagag taaaaaattg tacttggcgg ataatgcctt tagcggctta
     4441 actgtgccct ccatggaaaa atcagtcaag atatccacat gtgtttttag taaacaaatt
     4501 ttgggaccta atgcttcaac taactccagt aattccttgg tggtacgaac atccaatgaa
     4561 gcacacaagt ttgtttgctt ttcgtgcatg atattaaata gcttggcagc aacaggacta
     4621 ggatgagtag cagcacgttc cttatatgta gctttcgaca tgatttatct tcgtttcctg
     4681 caggtttttg ttctgtgcag ttgggttaag aatactgggc aatttcatgt ttcttcaaca
     4741 ctacatatgc gtatatatac caatctaagt ctgtgctcct tccttcgttc ttccttctgt
     4801 tcggagatta ccgaatcaaa aaaatttcaa ggaaaccgaa atcaaaaaaa agaataaaaa
     4861 aaaaatgatg aattgaaaag gtggtatggt gcactctcag tacaatctgc tctgatgccg
     4921 catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
     4981 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
     5041 ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt
     5101 tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa
     5161 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     5221 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     5281 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     5341 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     5401 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     5461 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     5521 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     5581 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     5641 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     5701 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     5761 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     5821 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     5881 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     5941 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     6001 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     6061 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     6121 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     6181 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     6241 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     6301 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     6361 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     6421 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     6481 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     6541 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     6601 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaac
//
