LOCUS       sequence-461105-        7434 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Visualization of nucleus in mammalian cells using mGold2s.
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-mGold2s-CAF1-C-10
FEATURES             Location/Qualifiers
     source          1..7434
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    691..812
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(728..747)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     782..801
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(819..1274)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(956..977)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1168..1187
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1301..1405
                     /gene="bla"
                     /label=AmpR promoter
     promoter        1407..1764
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        1407..1598
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(1430..1450)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      1615..1750
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             1799..2593
                     /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             1799..2593
                     /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(1853..1872)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2463..2482
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(2781..2800)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    2825..2872
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3201..3789
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3690..3709
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        3964..4267
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4268..4471
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4422..4442
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             join(4516..4518,4519..4521,4522..5232)
                     /codon_start=1
                     /product="photostable derivative of mVenus (Lee et al., 
                     2020)"
                     /label=mGold
                     /note="mammalian codon-optimized"
                     /translation="MASKGEELFTGVVPILVELDGDINGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTSLGYGLQCFARYPDNMKRHDFFKSAMPEGYVQERTIFFEDD
                     GNYKTRAEVKFEGGTLVNRVELKGIDFKEDGNILGHKLEYNYNCHNVFITADRQQNGIK
                     VNFKIRHNIEVGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLSMDELYK"
     primer_bind     complement(4822..4841)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5169..5190
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    7051..7062
                     /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)"
ORIGIN
        1 gtgaatggga gcaaggtgat catccatgaa ttccaggagc aatgccgcag gggcctgctg
       61 acccttccaa gtcccacccc acacctgcag atgcccaact tggaggatgc tgttgctgtg
      121 ccatccaagg ccaggctgaa gcgccttatc tctgagaact ctgcctatga gaagcggccc
      181 aacttccgta tgtgctggta tgtgcaccca gaggtgctca agagctttgg ccaggagtgc
      241 ctgccagtgc cctgccagtg gacctacatc accaccatgc cctcagcacc cagggaagac
      301 agtggcagtg cttctactga ggggcctggc cagagcacac ctatgctact caagcggaag
      361 ccagcggcca ccatgtgcat cacgcagttt atgaagaagc ggcgctatga tggacaggtt
      421 gggtctgggg acatggatgg cttccaggca gacacagaag aggatgaaga ggatgacaca
      481 gactgtatga tcatagatgt gccagatgtt gggagtgacg tgtcagaagc tcccatacct
      541 gctcccacac tttgcaagtg aggatccacc ggatctagat aactgatcat aatcagccat
      601 accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg
      661 aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac
      721 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
      781 tgtggtttgt ccaaactcat caatgtatct taacgcgtaa attgtaagcg ttaatatttt
      841 gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat
      901 cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt
      961 ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt
     1021 ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag
     1081 gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg
     1141 aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc
     1201 gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc
     1261 gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     1321 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     1381 tcaataatat tgaaaaagga agagtcctga ggcggaaaga accagctgtg gaatgtgtgt
     1441 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     1501 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     1561 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     1621 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     1681 tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt
     1741 tttggaggcc taggcttttg caaagatcga tcaagagaca ggatgaggat cgtttcgcat
     1801 gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg
     1861 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc
     1921 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca
     1981 agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct
     2041 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga
     2101 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg
     2161 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat
     2221 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga
     2281 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg
     2341 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg
     2401 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat
     2461 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct
     2521 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga
     2581 cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg
     2641 ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt
     2701 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc
     2761 caccctaggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct
     2821 atgacggcaa taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc
     2881 ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa
     2941 tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg
     3001 ctcgcagcca acgtcggggc ggcaggccct gccatagcct caggttactc atatatactt
     3061 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     3121 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     3181 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     3241 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     3301 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     3361 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     3421 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     3481 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     3541 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     3601 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     3661 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     3721 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     3781 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     3841 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccatg
     3901 cattagttat taatagtaat caattacggg gtcattagtt catagcccat atatggagtt
     3961 ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc
     4021 attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg
     4081 tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat
     4141 gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca
     4201 gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat
     4261 taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg
     4321 gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca
     4381 acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg
     4441 tgtacggtgg gaggtctata taagcagagc tggtttagtg aaccgtcaga tccgctagcg
     4501 ctaccggtgg ctaccatggc gagcaagggc gaggagctgt tcaccggggt ggtgcccatc
     4561 ctggtcgagc tggacggcga cataaacggc cacaagttca gcgtgtccgg cgagggcgag
     4621 ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc
     4681 gtgccctggc ccaccctcgt gaccagcctg ggctacggcc tgcagtgctt cgcccgctac
     4741 cccgacaaca tgaagcggca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag
     4801 gagcgcacca tcttcttcga ggacgacggc aactacaaga cccgcgccga ggtgaagttt
     4861 gagggcggca ccctggtgaa ccgcgtcgag ctgaagggca tcgacttcaa ggaggacggc
     4921 aacatcctgg ggcacaagct ggagtacaac tacaactgcc acaacgtctt tatcaccgcc
     4981 gacaggcagc agaacggcat caaggtcaac ttcaagatcc gccacaacat cgaggtcggc
     5041 ggcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg
     5101 ctgcccgaca accactacct gagctaccag tccaagctga gcaaagaccc caacgagaag
     5161 cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct cagcatggac
     5221 gagctgtaca agtccggact cagatctggc agcggtggag gcttggagga gccggaggcc
     5281 gcgacgcgga cagccgcggc cgtggattgc aaagacagac ccgggttccc agtgaagagg
     5341 ttaatacaag cccgcttgcc cttcaaacgc ctgaaccttg ttcccaagga gaaagttgag
     5401 gaagacacat caccaaaagc agctgtggaa agcaaagtcc cggatctaca attgtctttg
     5461 ggcacttttg aaagccagtg tcacacaggc tctcacgtag gcttgagtac aaagttggtc
     5521 ggtggccagg gccccattga tagcttcttg agggcaacaa tcaagccagt accgagtgtg
     5581 gtcattatcg acctgacgga gaactgcagt gacattccag acagccctga gggccacagt
     5641 gaactaagtc ctgacactgc tggtgtggtc accacagtgg aaggagctgc caagcagcag
     5701 gagcactctg cagcagagct gtgtctccta gagacccctt cagacatcac ttgtcacatg
     5761 gaagaggagc ctgggagtcc aggagaccca aagaggacgg gtgactgtca ggctggttca
     5821 ctacagagct gtccggagct gaccccaggt tctaggacat gccccaccaa ggagttatca
     5881 agctggagca aggcagggga tctcctattt atagagaaag tgcctgtggt ggtgctagag
     5941 gacatcctgg ccactaagcc ctccattgcc tctcttccca tgatgtccct ggataggagt
     6001 gtcacttcag agagtgaaat actggagtcc tgccccgagg atgactccat actgagccac
     6061 tcctccacca actcctcttc tccaaccagc tctcctgagg gaccatccac acccccagaa
     6121 catcggggtg ggagaagctc cccctccacc ccagcctgca gagttgccaa gaactttgtc
     6181 aaaggctcca cagagaaggg caggagcaaa ctgcacagag atagagaaca gcagagggaa
     6241 gagaaggaaa agctgaggga agagatacga cgagccaagg aggaggcacg gaagaagaag
     6301 gaggaggaga aggagctgaa ggaaaaggag cggagggaaa agcgggagaa agatgagaag
     6361 gagaaggcgg agaagcagcg gctaaaggag gaaaaacgca aggagcggca ggaagctctg
     6421 gaggctaaac tggaggagaa gaggaagaag gaggaagaga agcggttacg ggaggaagag
     6481 aagcggttac gggaagaaga gaagcgcatt aaggcagaaa aggcagagat cactcgattc
     6541 ttccagaaac cgaagactcc gcaggctccc aagaccctgg ccggttcctg tggtaagttt
     6601 gccccctttg aaatcaaaga acacatggtg ctggctccac ggtgccgagc tgccttggat
     6661 caagacctgt gcgatcagct ggaccagctc ctgcagcagc agagtgtggc aagcaccttc
     6721 ctcagtgacc taaaaagccg gctgcccttg agatcggggc ccacccgggt ctgtggccat
     6781 gacacagaca tcatgaacag ggatgtggtc attgtggaga gcagcaaggt ggacggtgtg
     6841 tctgaaagga agaagtttgg ccgcatgaag cttctgcagt tctcagagaa ccaccggcca
     6901 gcatactggg ggacgtggaa caagaagaca gccatcatcc gcccccggaa cccctgggcc
     6961 caggacaaag accttcttga ctatgaggta gacagtgacg atgagtggga agaggaggaa
     7021 cctggggagt ctttgtccca cagtgagggg gatgaggatg acgatgtggg ggaagatgag
     7081 gatgaggacg atggcttctt tgtgccccat gggtacttat ctgaagatga aggcgtgact
     7141 gaggagtgtg cagaccctga gaaccacaaa gtacaccaga agctgaaggc caaggagtgg
     7201 gatgagctcc tagccaaggg caagcgcttt cgtgtgctgc agcctgtgca cgtgggatgt
     7261 gtatgggcat cagaggcagc caactgcacg agttccgact tgaagctgct gcagcagttc
     7321 actgcatgcc tgctggatgt ggcatcccct gatgagccag aacccggggc ctccaggagg
     7381 gaaaagaggg accagcatat cctggcccag ttgcttccgc tgctacatgg caat
//
