LOCUS       sequence-463669-        7671 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of mGold2t in yeast cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7671)
  AUTHORS   Lee J, Lai S, Yang S, Zhao S, Blanco FA, Lyons AC, Merino-Urteaga R,
            Ahrens JF, Nguyen NA, Liu H, Liu Z, Lambert GG, Shaner NC, Chen L, 
            Tolias KF, Zhang J, Ha T, St-Pierre F
  TITLE     Bright and photostable yellow fluorescent proteins for extended 
            imaging.
  JOURNAL   Nat Commun. 2025 Apr 4;16(1):3241. doi: 10.1038/s41467-025-58223-5.
  PUBMED    40185748
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: "Nat 
            Commun."; date: "2025-04-4"; volume: "16(1):3241. doi"; pages: " 
            10.1038/s41467-025-58223-5"
COMMENT     Sequence Label: pJL-mGold2t
FEATURES             Location/Qualifiers
     source          1..7671
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(23..41)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        109..213
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(214..282,283..1074)
                     /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(432..451)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1245..1833
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1734..1753
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1987..2004
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2121..2142
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2157..2162,2163..2180,2181..2187)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2195..2211
                     /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     2200..2222
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2219..2235
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2219..2235
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     rep_origin      complement(2265..2720)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2402..2423)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2614..2633
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2748..3414
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase; also known as the TDH3 promoter"
     primer_bind     3284..3307
                     /label=GPDpro-F
                     /note="S. cerevisiae GPD promoter, forward primer"
     CDS             join(3421..3423,3424..3426,3427..4140)
                     /codon_start=1
                     /product="photostable derivative of mVenus (Lee et al., 
                     2020)"
                     /label=mGold
                     /note="mammalian codon-optimized"
                     /translation="MASKGEELFTGVVPILVELDGDINGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTSLGYGLQCFARYPDNMKRHDFFKSAMPEGYVQERTIFFEDD
                     GNYKTRAEVKFEGGTLVNRVELKGIDFKEDGNILGHKLEYNYNCHNVFITADRQQNGIK
                     VNFKIRHNIEVGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQTKLSKDPNEKRDHMVLL
                     EFVTAAGITLSMDELYK"
     primer_bind     complement(3727..3746)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4074..4095
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(4142..4158)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4142..4158)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     terminator      4159..4346
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(4375..4394)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4376..4394)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      4542..4729
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(4741..4758)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4741..4757)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4750..4772)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5193..6539
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     CDS             complement(6650..7453)
                     /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(7454..7669)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
ORIGIN
        1 gctaattctt gaagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg
       61 ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
      121 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
      181 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
      241 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
      301 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
      361 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
      421 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
      481 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
      541 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
      601 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
      661 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
      721 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
      781 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
      841 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
      901 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
      961 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     1021 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     1081 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     1141 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     1201 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     1261 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     1321 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     1381 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     1441 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     1501 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     1561 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     1621 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     1681 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     1741 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     1801 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     1861 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     1921 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     1981 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc
     2041 cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg
     2101 ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta
     2161 cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca
     2221 ggaaacagct atgaccatga ttacgccaag cttaccgcat caggaaattg taagcgttaa
     2281 tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc
     2341 cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt
     2401 tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa
     2461 aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg
     2521 gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg
     2581 acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc
     2641 tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa
     2701 tgcgccgcta cagggcgcgt ccattcgcgc tagccagttc gagtttatca ttatcaatac
     2761 tgccatttca aagaatacgt aaataattaa tagtagtgat tttcctaact ttatttagtc
     2821 aaaaaattag ccttttaatt ctgctgtaac ccgtacatgc ccaaaatagg gggcgggtta
     2881 cacagaatat ataacatcgt aggtgtctgg gtgaacagtt tattcctggc atccactaaa
     2941 tataatggag cccgcttttt aagctggcat ccagaaaaaa aaagaatccc agcaccaaaa
     3001 tattgttttc ttcaccaacc atcagttcat aggtccattc tcttagcgca actacagaga
     3061 acaggggcac aaacaggcaa aaaacgggca caacctcaat ggagtgatgc aacctgcctg
     3121 gagtaaatga tgacacaagg caattgaccc acgcatgtat ctatctcatt ttcttacacc
     3181 ttctattacc ttctgctctc tctgatttgg aaaaagctga aaaaaaaggt tgaaaccagt
     3241 tccctgaaat tattccccta cttgactaat aagtatataa agacggtagg tattgattgt
     3301 aattctgtaa atctatttct taaacttctt aaattctact tttatagtta gtcttttttt
     3361 tagttttaaa acaccaagaa cttagtttcg aataaacaca cataaacaaa caaaagatct
     3421 atggcgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
     3481 ggcgacataa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac
     3541 ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc
     3601 ctcgtgacca gcctgggcta cggcctgcag tgcttcgccc gctaccccga caacatgaag
     3661 cggcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc
     3721 ttcgaggacg acggcaacta caagacccgc gccgaggtga agtttgaggg cggcaccctg
     3781 gtgaaccgcg tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac
     3841 aagctggagt acaactacaa ctgccacaac gtctttatca ccgccgacag gcagcagaac
     3901 ggcatcaagg tcaacttcaa gatccgccac aacatcgagg tcggcggcgt gcagctcgcc
     3961 gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac
     4021 tacctgagct accagaccaa gctgagcaaa gaccccaacg agaagcgcga tcacatggtc
     4081 ctgctggagt tcgtgaccgc cgccgggatc actctcagca tggacgagct gtacaagtaa
     4141 ggatccacta gttctagagc gaatttctta tgatttatga tttttattat taaataagtt
     4201 ataaaaaaaa taagtgtata caaattttaa agtgactctt aggttttaaa acgaaaattc
     4261 ttattcttga gtaactcttt cctgtaggtc aggttgcttt ctcaggtata gtatgaggtc
     4321 gctcttattg accacacctc taccggctcg agggggggcc cggtacccaa ttcgccctat
     4381 agtgagtcgt attacgcgcg catccagctt tggacttctt cgccagaggt ttggtcaagt
     4441 ctccaatcaa ggttgtcggc ttgtctacct tgccagaaat ttacgaaaag atggaaaagg
     4501 gtcaaatcgt tggtagatac gttgttgaca cttctaaata agcgaatttc ttatgattta
     4561 tgatttttat tattaaataa gttataaaaa aaataagtgt atacaaattt taaagtgact
     4621 cttaggtttt aaaacgaaaa ttcttattct tgagtaactc tttcctgtag gtcaggttgc
     4681 tttctcaggt atagcatgag gtcgctctta ttgaccacac ctctaccggc atgccaattc
     4741 actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg
     4801 ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg
     4861 cccttcccaa cagttgcgca gcctgaatgg cgaatggcgc ctgatgcggt attttctcct
     4921 tacgcatctg tgcggtattt cacaccgcat aatcggatcg tacttgttac ccatcattga
     4981 attttgaaca tccgaacctg ggagttttcc ctgaaacaga tagtatattt gaacctgtat
     5041 aataatatat agtctagcgc tttacggaag acaatgtatg tatttcggtt cctggagaaa
     5101 ctattgcatc tattgcatag gtaatcttgc acgtcgcatc cccggttcat tttctgcgtt
     5161 tccatcttgc acttcaatag catatctttg ttaacgaagc atctgtgctt cattttgtag
     5221 aacaaaaatg caacgcgaga gcgctaattt ttcaaacaaa gaatctgagc tgcattttta
     5281 cagaacagaa atgcaacgcg aaagcgctat tttaccaacg aagaatctgt gcttcatttt
     5341 tgtaaaacaa aaatgcaacg cgagagcgct aatttttcaa acaaagaatc tgagctgcat
     5401 ttttacagaa cagaaatgca acgcgagagc gctattttac caacaaagaa tctatacttc
     5461 ttttttgttc tacaaaaatg catcccgaga gcgctatttt tctaacaaag catcttagat
     5521 tacttttttt ctcctttgtg cgctctataa tgcagtctct tgataacttt ttgcactgta
     5581 ggtccgttaa ggttagaaga aggctacttt ggtgtctatt ttctcttcca taaaaaaagc
     5641 ctgactccac ttcccgcgtt tactgattac tagcgaagct gcgggtgcat tttttcaaga
     5701 taaaggcatc cccgattata ttctataccg atgtggattg cgcatacttt gtgaacagaa
     5761 agtgatagcg ttgatgattc ttcattggtc agaaaattat gaacggtttc ttctattttg
     5821 tctctatata ctacgtatag gaaatgttta cattttcgta ttgttttcga ttcactctat
     5881 gaatagttct tactacaatt tttttgtcta aagagtaata ctagagataa acataaaaaa
     5941 tgtagaggtc gagtttagat gcaagttcaa ggagcgaaag gtggatgggt aggttatata
     6001 gggatatagc acagagatat atagcaaaga gatacttttg agcaatgttt gtggaagcgg
     6061 tattcgcaat attttagtag ctcgttacag tccggtgcgt ttttggtttt ttgaaagtgc
     6121 gtcttcagag cgcttttggt tttcaaaagc gctctgaagt tcctatactt tctagctaga
     6181 gaataggaac ttcggaatag gaacttcaaa gcgtttccga aaacgagcgc ttccgaaaat
     6241 gcaacgcgag ctgcgcacat acagctcact gttcacgtcg cacctatatc tgcgtgttgc
     6301 ctgtatatat atatacatga gaagaacggc atagtgcgtg tttatgctta aatgcgtact
     6361 tatatgcgtc tatttatgta ggatgaaagg tagtctagta cctcctgtga tattatccca
     6421 ttccatgcgg ggtatcgtat gcttccttca gcactaccct ttagctgttc tatatgctgc
     6481 cactcctcaa ttggattagt ctcatccttc aatgctatca tttcctttga tattggatcg
     6541 atccgatgat aagctgtcaa acatgagaat tgggtaataa ctgatataat taaattgaag
     6601 ctctaatttg tgagtttagt atacatgcat ttacttataa tacagttttt tagttttgct
     6661 ggccgcatct tctcaaatat gcttcccagc ctgcttttct gtaacgttca ccctctacct
     6721 tagcatccct tccctttgca aatagtcctc ttccaacaat aataatgtca gatcctgtag
     6781 agaccacatc atccacggtt ctatactgtt gacccaatgc gtctcccttg tcatctaaac
     6841 ccacaccggg tgtcataatc aaccaatcgt aaccttcatc tcttccaccc atgtctcttt
     6901 gagcaataaa gccgataaca aaatctttgt cgctcttcgc aatgtcaaca gtacccttag
     6961 tatattctcc agtagatagg gagcccttgc atgacaattc tgctaacatc aaaaggcctc
     7021 taggttcctt tgttacttct tctgccgcct gcttcaaacc gctaacaata cctgggccca
     7081 ccacaccgtg tgcattcgta atgtctgccc attctgctat tctgtataca cccgcagagt
     7141 actgcaattt gactgtatta ccaatgtcag caaattttct gtcttcgaag agtaaaaaat
     7201 tgtacttggc ggataatgcc tttagcggct taactgtgcc ctccatggaa aaatcagtca
     7261 agatatccac atgtgttttt agtaaacaaa ttttgggacc taatgcttca actaactcca
     7321 gtaattcctt ggtggtacga acatccaatg aagcacacaa gtttgtttgc ttttcgtgca
     7381 tgatattaaa tagcttggca gcaacaggac taggatgagt agcagcacgt tccttatatg
     7441 tagctttcga catgatttat cttcgtttcc tgcatgtttt tgttctgtgc agttgggtta
     7501 agaatactgg gcaatttcat gtttcttcaa cactacatat gcgtatatat accaatctaa
     7561 gtctgtgctc cttccttcgt tcttccttct gttcggagat taccgaatca aaaaaatttc
     7621 aaggaaaccg aaatcaaaaa aaagaataaa aaaaaaatga tgaattgaaa a
//
