LOCUS       sequence-461119-        7434 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Visualization of nucleus in mammalian cells using mGold2t.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7434)
  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 7434)
  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: pCMV-mGold2t-CAF1-C-10
FEATURES             Location/Qualifiers
     source          1..7434
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        92..395
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        396..599
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     550..570
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             join(644..646,647..649,650..1360)
                     /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(950..969)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1297..1318
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    3179..3190
                     /label=ZF binding site
                     /bound_moiety="CCR5TC zinc-finger domain"
                     /note="target sequence for the CCR5TC zinc-finger domain 
                     from the CCR5-224 (+) zinc-finger nuclease (Perez et al., 
                     2008; Gross et al., 2013)"
     polyA_signal    4253..4374
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4290..4309)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4344..4363
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4381..4836)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4518..4539)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4730..4749
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4863..4967
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4969..5326
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4969..5160
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4992..5012)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5177..5312
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5361..6155
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             5361..6155
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5415..5434)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6025..6044
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6343..6362)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6387..6434
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6763..7351
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7252..7271
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttctgtggat aaccgtatta ccgccatgca ttagttatta atagtaatca attacggggt
       61 cattagttca tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc
      121 ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag
      181 taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc
      241 acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg
      301 gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc
      361 agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca
      421 atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca
      481 atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg
      541 ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctg
      601 gtttagtgaa ccgtcagatc cgctagcgct accggtggct accatggcga gcaagggcga
      661 ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgaca taaacggcca
      721 caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa
      781 gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga ccagcctggg
      841 ctacggcctg cagtgcttcg cccgctaccc cgacaacatg aagcggcacg acttcttcaa
      901 gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcgagg acgacggcaa
      961 ctacaagacc cgcgccgagg tgaagtttga gggcggcacc ctggtgaacc gcgtcgagct
     1021 gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta
     1081 caactgccac aacgtcttta tcaccgccga caggcagcag aacggcatca aggtcaactt
     1141 caagatccgc cacaacatcg aggtcggcgg cgtgcagctc gccgaccact accagcagaa
     1201 cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga gctaccagac
     1261 caagctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac
     1321 cgccgccggg atcactctca gcatggacga gctgtacaag tccggactca gatctggcag
     1381 cggtggaggc ttggaggagc cggaggccgc gacgcggaca gccgcggccg tggattgcaa
     1441 agacagaccc gggttcccag tgaagaggtt aatacaagcc cgcttgccct tcaaacgcct
     1501 gaaccttgtt cccaaggaga aagttgagga agacacatca ccaaaagcag ctgtggaaag
     1561 caaagtcccg gatctacaat tgtctttggg cacttttgaa agccagtgtc acacaggctc
     1621 tcacgtaggc ttgagtacaa agttggtcgg tggccagggc cccattgata gcttcttgag
     1681 ggcaacaatc aagccagtac cgagtgtggt cattatcgac ctgacggaga actgcagtga
     1741 cattccagac agccctgagg gccacagtga actaagtcct gacactgctg gtgtggtcac
     1801 cacagtggaa ggagctgcca agcagcagga gcactctgca gcagagctgt gtctcctaga
     1861 gaccccttca gacatcactt gtcacatgga agaggagcct gggagtccag gagacccaaa
     1921 gaggacgggt gactgtcagg ctggttcact acagagctgt ccggagctga ccccaggttc
     1981 taggacatgc cccaccaagg agttatcaag ctggagcaag gcaggggatc tcctatttat
     2041 agagaaagtg cctgtggtgg tgctagagga catcctggcc actaagccct ccattgcctc
     2101 tcttcccatg atgtccctgg ataggagtgt cacttcagag agtgaaatac tggagtcctg
     2161 ccccgaggat gactccatac tgagccactc ctccaccaac tcctcttctc caaccagctc
     2221 tcctgaggga ccatccacac ccccagaaca tcggggtggg agaagctccc cctccacccc
     2281 agcctgcaga gttgccaaga actttgtcaa aggctccaca gagaagggca ggagcaaact
     2341 gcacagagat agagaacagc agagggaaga gaaggaaaag ctgagggaag agatacgacg
     2401 agccaaggag gaggcacgga agaagaagga ggaggagaag gagctgaagg aaaaggagcg
     2461 gagggaaaag cgggagaaag atgagaagga gaaggcggag aagcagcggc taaaggagga
     2521 aaaacgcaag gagcggcagg aagctctgga ggctaaactg gaggagaaga ggaagaagga
     2581 ggaagagaag cggttacggg aggaagagaa gcggttacgg gaagaagaga agcgcattaa
     2641 ggcagaaaag gcagagatca ctcgattctt ccagaaaccg aagactccgc aggctcccaa
     2701 gaccctggcc ggttcctgtg gtaagtttgc cccctttgaa atcaaagaac acatggtgct
     2761 ggctccacgg tgccgagctg ccttggatca agacctgtgc gatcagctgg accagctcct
     2821 gcagcagcag agtgtggcaa gcaccttcct cagtgaccta aaaagccggc tgcccttgag
     2881 atcggggccc acccgggtct gtggccatga cacagacatc atgaacaggg atgtggtcat
     2941 tgtggagagc agcaaggtgg acggtgtgtc tgaaaggaag aagtttggcc gcatgaagct
     3001 tctgcagttc tcagagaacc accggccagc atactggggg acgtggaaca agaagacagc
     3061 catcatccgc ccccggaacc cctgggccca ggacaaagac cttcttgact atgaggtaga
     3121 cagtgacgat gagtgggaag aggaggaacc tggggagtct ttgtcccaca gtgaggggga
     3181 tgaggatgac gatgtggggg aagatgagga tgaggacgat ggcttctttg tgccccatgg
     3241 gtacttatct gaagatgaag gcgtgactga ggagtgtgca gaccctgaga accacaaagt
     3301 acaccagaag ctgaaggcca aggagtggga tgagctccta gccaagggca agcgctttcg
     3361 tgtgctgcag cctgtgcacg tgggatgtgt atgggcatca gaggcagcca actgcacgag
     3421 ttccgacttg aagctgctgc agcagttcac tgcatgcctg ctggatgtgg catcccctga
     3481 tgagccagaa cccggggcct ccaggaggga aaagagggac cagcatatcc tggcccagtt
     3541 gcttccgctg ctacatggca atgtgaatgg gagcaaggtg atcatccatg aattccagga
     3601 gcaatgccgc aggggcctgc tgacccttcc aagtcccacc ccacacctgc agatgcccaa
     3661 cttggaggat gctgttgctg tgccatccaa ggccaggctg aagcgcctta tctctgagaa
     3721 ctctgcctat gagaagcggc ccaacttccg tatgtgctgg tatgtgcacc cagaggtgct
     3781 caagagcttt ggccaggagt gcctgccagt gccctgccag tggacctaca tcaccaccat
     3841 gccctcagca cccagggaag acagtggcag tgcttctact gaggggcctg gccagagcac
     3901 acctatgcta ctcaagcgga agccagcggc caccatgtgc atcacgcagt ttatgaagaa
     3961 gcggcgctat gatggacagg ttgggtctgg ggacatggat ggcttccagg cagacacaga
     4021 agaggatgaa gaggatgaca cagactgtat gatcatagat gtgccagatg ttgggagtga
     4081 cgtgtcagaa gctcccatac ctgctcccac actttgcaag tgaggatcca ccggatctag
     4141 ataactgatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct
     4201 cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt
     4261 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     4321 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttaacgcgt
     4381 aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat
     4441 tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga
     4501 tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca
     4561 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct
     4621 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc
     4681 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag
     4741 cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca
     4801 cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt cggggaaatg
     4861 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     4921 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtcct gaggcggaaa
     4981 gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
     5041 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
     5101 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
     5161 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     5221 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt
     5281 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaagatc gatcaagaga
     5341 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg
     5401 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
     5461 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
     5521 ccggtgccct gaatgaactg caagacgagg cagcgcggct atcgtggctg gccacgacgg
     5581 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
     5641 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
     5701 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
     5761 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
     5821 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
     5881 tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     5941 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     6001 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
     6061 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     6121 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac
     6181 cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga
     6241 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     6301 tctcatgctg gagttcttcg cccaccctag ggggaggcta actgaaacac ggaaggagac
     6361 aataccggaa ggaacccgcg ctatgacggc aataaaaaga cagaataaaa cgcacggtgt
     6421 tgggtcgttt gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc
     6481 accgagaccc cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc
     6541 aagttcgggt gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc
     6601 ctcaggttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     6661 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     6721 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     6781 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     6841 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     6901 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     6961 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     7021 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     7081 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     7141 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     7201 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     7261 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     7321 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     7381 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctga
//
