LOCUS       sequence-461104-        5778 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Visualization of actin in mammalian cells using mGold2t.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5778)
  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 5778)
  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-Actin-C-18
FEATURES             Location/Qualifiers
     source          1..5778
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      547..556
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(553..555,556..558,559..1269)
                     /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(859..878)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1206..1227
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2578..2699
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2615..2634)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2669..2688
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2706..3161)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2843..2864)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3055..3074
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3188..3292
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3317..3337)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3361..3571
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        3383..3579
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3430..3565
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3614..4408
                     /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             3614..4408
                     /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(3668..3687)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4278..4297
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4596..4615)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4640..4687
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5016..5604
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5505..5524
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      541 ccggtcgcca ccatggcgag caagggcgag gagctgttca ccggggtggt gcccatcctg
      601 gtcgagctgg acggcgacat aaacggccac aagttcagcg tgtccggcga gggcgagggc
      661 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
      721 ccctggccca ccctcgtgac cagcctgggc tacggcctgc agtgcttcgc ccgctacccc
      781 gacaacatga agcggcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
      841 cgcaccatct tcttcgagga cgacggcaac tacaagaccc gcgccgaggt gaagtttgag
      901 ggcggcaccc tggtgaaccg cgtcgagctg aagggcatcg acttcaagga ggacggcaac
      961 atcctggggc acaagctgga gtacaactac aactgccaca acgtctttat caccgccgac
     1021 aggcagcaga acggcatcaa ggtcaacttc aagatccgcc acaacatcga ggtcggcggc
     1081 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1141 cccgacaacc actacctgag ctaccagacc aagctgagca aagaccccaa cgagaagcgc
     1201 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcag catggacgag
     1261 ctgtacaagt ccggactcag atctggcagc ggtggaggca gcgcatccgg cggaagcgga
     1321 agcgatgatg atatcgccgc gctcgtcgtc gacaacggct ccggcatgtg caaggccggc
     1381 ttcgcgggcg acgatgcccc ccgggccgtc ttcccctcca tcgtggggcg ccccaggcac
     1441 cagggcgtga tggtgggcat gggtcagaag gattcctatg tgggcgacga ggcccagagc
     1501 aagagaggca tcctcaccct gaagtacccc atcgagcacg gcatcgtcac caactgggac
     1561 gacatggaga aaatctggca ccacaccttc tacaatgagc tgcgtgtggc tcccgaggag
     1621 caccccgtgc tgctgaccga ggcccccctg aaccccaagg ccaaccgcga gaagatgacc
     1681 cagatcatgt ttgagacctt caacacccca gccatgtacg ttgctatcca ggctgtgcta
     1741 tccctgtacg cctctggccg taccactggc atcgtgatgg actccggtga cggggtcacc
     1801 cacactgtgc ccatctacga ggggtatgcc ctcccccatg ccatcctgcg tctggacctg
     1861 gctggccggg acctgactga ctacctcatg aagatcctca ccgagcgcgg ctacagcttc
     1921 accaccacgg ccgagcggga aatcgtgcgt gacattaagg agaagctgtg ctacgtcgcc
     1981 ctggacttcg agcaagagat ggccacggct gcttccagct cctccctgga gaagagctac
     2041 gagctgcctg acggccaggt catcaccatt ggcaatgagc ggttccgctg ccctgaggca
     2101 ctcttccagc cttccttcct gggcatggag tcctgtggca tccacgaaac taccttcaac
     2161 tccatcatga agtgtgacgt ggacatccgc aaagacctgt acgccaacac agtgctgtct
     2221 ggcggcacca ccatgtaccc tggcattgcc gacaggatgc agaaggagat cactgccctg
     2281 gcacccagca caatgaagat caagatcatt gctcctcctg agcgcaagta ctccgtgtgg
     2341 atcggcggct ccatcctggc ctcgctgtcc accttccagc agatgtggat cagcaagcag
     2401 gagtatgacg agtccggccc ctccatcgtc caccgcaaat gcttctaggg atccaccgga
     2461 tctagataac tgatcataat cagccatacc acatttgtag aggttttact tgctttaaaa
     2521 aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac
     2581 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     2641 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa
     2701 ggcgtaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag
     2761 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac
     2821 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga
     2881 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc
     2941 accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
     3001 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
     3061 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
     3121 caccacaccc gccgcgctta atgcgccgct acagggcgcg tcaggtggca cttttcgggg
     3181 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     3241 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtcctgaggc
     3301 ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     3361 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     3421 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     3481 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag
     3541 tagtgaggag gcttttttgg aggcctaggc ttttgcaaag atcgatcaag agacaggatg
     3601 aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt
     3661 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
     3721 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
     3781 cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
     3841 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
     3901 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat
     3961 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
     4021 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
     4081 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
     4141 gagcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat
     4201 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga
     4261 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg
     4321 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
     4381 ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa
     4441 gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg
     4501 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
     4561 ctggagttct tcgcccaccc tagggggagg ctaactgaaa cacggaagga gacaataccg
     4621 gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg tgttgggtcg
     4681 tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga
     4741 ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg
     4801 ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat agcctcaggt
     4861 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     4921 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     4981 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     5041 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     5101 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     5161 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     5221 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     5281 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     5341 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     5401 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     5461 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     5521 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     5581 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     5641 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     5701 cgtattaccg ccatgcatta gttattaata gtaatcaatt acggggtcat tagttcatag
     5761 cccatatatg gagttccg
//
