LOCUS       sequence-309931-        7671 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of mGold in yeast cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7671)
  AUTHORS   Lee J, Liu Z, Suzuki PH, Ahrens JF, Lai S, Lu X, Guan S, St-Pierre F
  TITLE     Versatile phenotype-activated cell sorting.
  JOURNAL   Sci Adv. 2020 Oct 23;6(43). pii: 6/43/eabb7438. doi: 
            10.1126/sciadv.abb7438. Print 2020 Oct.
  PUBMED    33097540
REFERENCE   2  (bases 1 to 7671)
  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: "Sci Adv."; 
            date: "2020-10-23"; volume: "6(43). pii: 6/43/eabb7438. doi"; pages:
            " 10.1126/sciadv.abb7438. Print 2020 Oct"
COMMENT     Sequence Label: pJL-mGold
FEATURES             Location/Qualifiers
     source          1..7671
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(7..9,10..12,13..726)
                     /codon_start=1
                     /product="photostable derivative of mVenus (Lee et al., 
                     2020)"
                     /label=mGold
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTSLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(52..73)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(313..332)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     660..681
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(728..744)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(728..744)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     terminator      745..932
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(961..980)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(962..980)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      1128..1315
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1327..1344)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1327..1343)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1336..1358)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1779..3125
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     CDS             complement(3236..4039)
                     /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(4040..4255)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(4280..4298)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4366..4470
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4471..4539,4540..5331)
                     /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(4689..4708)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5502..6090
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5991..6010
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6244..6261
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6378..6399
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6414..6419,6420..6437,6438..6444)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6452..6468
                     /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     6457..6479
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6476..6492
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6476..6492
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     rep_origin      complement(6522..6977)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6659..6680)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6871..6890
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        7005..7671
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase; also known as the TDH3 promoter"
     primer_bind     7541..7564
                     /label=GPDpro-F
                     /note="S. cerevisiae GPD promoter, forward primer"
ORIGIN
        1 agatctatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag
       61 ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc
      121 acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg
      181 cccaccctcg tgaccagcct gggctacggc ctgcagtgct tcgcccgcta ccccgaccac
      241 atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc
      301 atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac
      361 accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg
      421 gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcaccgc cgacaagcag
      481 aagaacggca tcaaggccaa cttcaagatc cgccacaaca tcgaggacgg cggcgtgcag
      541 ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac
      601 aaccactacc tgagctacca gtccaagctg agcaaagacc ccaacgagaa gcgcgatcac
      661 atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac
      721 aagtaaggat ccactagttc tagagcgaat ttcttatgat ttatgatttt tattattaaa
      781 taagttataa aaaaaataag tgtatacaaa ttttaaagtg actcttaggt tttaaaacga
      841 aaattcttat tcttgagtaa ctctttcctg taggtcaggt tgctttctca ggtatagtat
      901 gaggtcgctc ttattgacca cacctctacc ggctcgaggg ggggcccggt acccaattcg
      961 ccctatagtg agtcgtatta cgcgcgcatc cagctttgga cttcttcgcc agaggtttgg
     1021 tcaagtctcc aatcaaggtt gtcggcttgt ctaccttgcc agaaatttac gaaaagatgg
     1081 aaaagggtca aatcgttggt agatacgttg ttgacacttc taaataagcg aatttcttat
     1141 gatttatgat ttttattatt aaataagtta taaaaaaaat aagtgtatac aaattttaaa
     1201 gtgactctta ggttttaaaa cgaaaattct tattcttgag taactctttc ctgtaggtca
     1261 ggttgctttc tcaggtatag catgaggtcg ctcttattga ccacacctct accggcatgc
     1321 caattcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact
     1381 taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac
     1441 cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tggcgcctga tgcggtattt
     1501 tctccttacg catctgtgcg gtatttcaca ccgcataatc ggatcgtact tgttacccat
     1561 cattgaattt tgaacatccg aacctgggag ttttccctga aacagatagt atatttgaac
     1621 ctgtataata atatatagtc tagcgcttta cggaagacaa tgtatgtatt tcggttcctg
     1681 gagaaactat tgcatctatt gcataggtaa tcttgcacgt cgcatccccg gttcattttc
     1741 tgcgtttcca tcttgcactt caatagcata tctttgttaa cgaagcatct gtgcttcatt
     1801 ttgtagaaca aaaatgcaac gcgagagcgc taatttttca aacaaagaat ctgagctgca
     1861 tttttacaga acagaaatgc aacgcgaaag cgctatttta ccaacgaaga atctgtgctt
     1921 catttttgta aaacaaaaat gcaacgcgag agcgctaatt tttcaaacaa agaatctgag
     1981 ctgcattttt acagaacaga aatgcaacgc gagagcgcta ttttaccaac aaagaatcta
     2041 tacttctttt ttgttctaca aaaatgcatc ccgagagcgc tatttttcta acaaagcatc
     2101 ttagattact ttttttctcc tttgtgcgct ctataatgca gtctcttgat aactttttgc
     2161 actgtaggtc cgttaaggtt agaagaaggc tactttggtg tctattttct cttccataaa
     2221 aaaagcctga ctccacttcc cgcgtttact gattactagc gaagctgcgg gtgcattttt
     2281 tcaagataaa ggcatccccg attatattct ataccgatgt ggattgcgca tactttgtga
     2341 acagaaagtg atagcgttga tgattcttca ttggtcagaa aattatgaac ggtttcttct
     2401 attttgtctc tatatactac gtataggaaa tgtttacatt ttcgtattgt tttcgattca
     2461 ctctatgaat agttcttact acaatttttt tgtctaaaga gtaatactag agataaacat
     2521 aaaaaatgta gaggtcgagt ttagatgcaa gttcaaggag cgaaaggtgg atgggtaggt
     2581 tatataggga tatagcacag agatatatag caaagagata cttttgagca atgtttgtgg
     2641 aagcggtatt cgcaatattt tagtagctcg ttacagtccg gtgcgttttt ggttttttga
     2701 aagtgcgtct tcagagcgct tttggttttc aaaagcgctc tgaagttcct atactttcta
     2761 gctagagaat aggaacttcg gaataggaac ttcaaagcgt ttccgaaaac gagcgcttcc
     2821 gaaaatgcaa cgcgagctgc gcacatacag ctcactgttc acgtcgcacc tatatctgcg
     2881 tgttgcctgt atatatatat acatgagaag aacggcatag tgcgtgttta tgcttaaatg
     2941 cgtacttata tgcgtctatt tatgtaggat gaaaggtagt ctagtacctc ctgtgatatt
     3001 atcccattcc atgcggggta tcgtatgctt ccttcagcac taccctttag ctgttctata
     3061 tgctgccact cctcaattgg attagtctca tccttcaatg ctatcatttc ctttgatatt
     3121 ggatcgatcc gatgataagc tgtcaaacat gagaattggg taataactga tataattaaa
     3181 ttgaagctct aatttgtgag tttagtatac atgcatttac ttataataca gttttttagt
     3241 tttgctggcc gcatcttctc aaatatgctt cccagcctgc ttttctgtaa cgttcaccct
     3301 ctaccttagc atcccttccc tttgcaaata gtcctcttcc aacaataata atgtcagatc
     3361 ctgtagagac cacatcatcc acggttctat actgttgacc caatgcgtct cccttgtcat
     3421 ctaaacccac accgggtgtc ataatcaacc aatcgtaacc ttcatctctt ccacccatgt
     3481 ctctttgagc aataaagccg ataacaaaat ctttgtcgct cttcgcaatg tcaacagtac
     3541 ccttagtata ttctccagta gatagggagc ccttgcatga caattctgct aacatcaaaa
     3601 ggcctctagg ttcctttgtt acttcttctg ccgcctgctt caaaccgcta acaatacctg
     3661 ggcccaccac accgtgtgca ttcgtaatgt ctgcccattc tgctattctg tatacacccg
     3721 cagagtactg caatttgact gtattaccaa tgtcagcaaa ttttctgtct tcgaagagta
     3781 aaaaattgta cttggcggat aatgccttta gcggcttaac tgtgccctcc atggaaaaat
     3841 cagtcaagat atccacatgt gtttttagta aacaaatttt gggacctaat gcttcaacta
     3901 actccagtaa ttccttggtg gtacgaacat ccaatgaagc acacaagttt gtttgctttt
     3961 cgtgcatgat attaaatagc ttggcagcaa caggactagg atgagtagca gcacgttcct
     4021 tatatgtagc tttcgacatg atttatcttc gtttcctgca tgtttttgtt ctgtgcagtt
     4081 gggttaagaa tactgggcaa tttcatgttt cttcaacact acatatgcgt atatatacca
     4141 atctaagtct gtgctccttc cttcgttctt ccttctgttc ggagattacc gaatcaaaaa
     4201 aatttcaagg aaaccgaaat caaaaaaaag aataaaaaaa aaatgatgaa ttgaaaagct
     4261 aattcttgaa gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata
     4321 ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     4381 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     4441 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     4501 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     4561 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     4621 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     4681 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     4741 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     4801 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     4861 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     4921 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     4981 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     5041 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     5101 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     5161 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     5221 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     5281 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     5341 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     5401 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     5461 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     5521 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     5581 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     5641 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     5701 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     5761 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     5821 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     5881 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     5941 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     6001 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     6061 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     6121 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     6181 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     6241 cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc
     6301 gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc
     6361 agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac
     6421 tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga
     6481 aacagctatg accatgatta cgccaagctt accgcatcag gaaattgtaa gcgttaatat
     6541 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga
     6601 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc
     6661 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
     6721 cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc
     6781 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
     6841 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag
     6901 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
     6961 gccgctacag ggcgcgtcca ttcgcgctag ccagttcgag tttatcatta tcaatactgc
     7021 catttcaaag aatacgtaaa taattaatag tagtgatttt cctaacttta tttagtcaaa
     7081 aaattagcct tttaattctg ctgtaacccg tacatgccca aaataggggg cgggttacac
     7141 agaatatata acatcgtagg tgtctgggtg aacagtttat tcctggcatc cactaaatat
     7201 aatggagccc gctttttaag ctggcatcca gaaaaaaaaa gaatcccagc accaaaatat
     7261 tgttttcttc accaaccatc agttcatagg tccattctct tagcgcaact acagagaaca
     7321 ggggcacaaa caggcaaaaa acgggcacaa cctcaatgga gtgatgcaac ctgcctggag
     7381 taaatgatga cacaaggcaa ttgacccacg catgtatcta tctcattttc ttacaccttc
     7441 tattaccttc tgctctctct gatttggaaa aagctgaaaa aaaaggttga aaccagttcc
     7501 ctgaaattat tcccctactt gactaataag tatataaaga cggtaggtat tgattgtaat
     7561 tctgtaaatc tatttcttaa acttcttaaa ttctactttt atagttagtc ttttttttag
     7621 ttttaaaaca ccaagaactt agtttcgaat aaacacacat aaacaaacaa a
//
