LOCUS       sequence-463668-        6081 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Visualization of tubulin in mammalian cells using mGold2t.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6081)
  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 6081)
  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-Tubulin-C-18
FEATURES             Location/Qualifiers
     source          1..6081
                     /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    2809..2930
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2846..2865)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2900..2919
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2937..3392)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3074..3095)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3286..3305
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3419..3523
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3525..3882
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3525..3716
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3548..3568)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3733..3868
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3917..4711
                     /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             3917..4711
                     /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(3971..3990)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4581..4600
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4899..4918)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4943..4990
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5319..5907
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5808..5827
                     /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 agcgtgcgtg agtgcatctc catccacgtt ggccaggctg gtgtccagat tggcaatgcc
     1381 tgctgggagc tctactgcct ggaacacggc atccagcccg atggccagat gccaagtgac
     1441 aagaccattg ggggaggaga tgactccttc aacaccttct tcagtgagac gggcgctggc
     1501 aagcacgtgc cccgggctgt gtttgtagac ttggaaccca cagtcattga tgaagttcgc
     1561 actggcacct accgccagct cttccaccct gagcagctca tcacaggcaa ggaagatgct
     1621 gccaataact atgcccgagg gcactacacc attggcaagg agatcattga ccttgtgttg
     1681 gaccgaattc gcaagctggc tgaccagtgc accggtcttc agggcttctt ggttttccac
     1741 agctttggtg ggggaactgg ttctgggttc acctccctgc tcatggaacg tctctcagtt
     1801 gattatggca agaagtccaa gctggagttc tccatttacc cagcacccca ggtttccaca
     1861 gctgtagttg agccctacaa ctccatcctc accacccaca ccaccctgga gcactctgat
     1921 tgtgccttca tggtagacaa tgaggccatc tatgacatct gtcgtagaaa cctcgatatc
     1981 gagcgcccaa cctacactaa ccttaaccgc cttattagcc agattgtgtc ctccatcact
     2041 gcttccctga gatttgatgg agccctgaat gttgacctga cagaattcca gaccaacctg
     2101 gtgccctacc cccgcatcca cttccctctg gccacatatg cccctgtcat ctctgctgag
     2161 aaagcctacc atgaacagct ttctgtagca gagatcacca atgcttgctt tgagccagcc
     2221 aaccagatgg tgaaatgtga ccctcgccat ggtaaataca tggcttgctg cctgttgtac
     2281 cgtggtgacg tggttcccaa agatgtcaat gctgccattg ccaccatcaa aaccaagcgc
     2341 agcatccagt ttgtggattg gtgccccact ggcttcaagg ttggcatcaa ctaccagcct
     2401 cccactgtgg tgcctggtgg agacctggcc aaggtacaga gagctgtgtg catgctgagc
     2461 aacaccacag ccattgctga ggcctgggct cgcctggacc acaagtttga cctgatgtat
     2521 gccaagcgtg cctttgttca ctggtacgtg ggtgagggga tggaggaagg cgagttttca
     2581 gaggcccgtg aagatatggc tgcccttgag aaggattatg aggaggttgg tgtggattct
     2641 gttgaaggag agggtgagga agaaggagag gaatactaag gatccaccgg atctagataa
     2701 ctgatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca
     2761 cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt
     2821 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     2881 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta acgcgtaaat
     2941 tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt
     3001 taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg
     3061 gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt
     3121 caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc
     3181 aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg
     3241 atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa
     3301 aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc
     3361 cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg
     3421 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     3481 ataaccctga taaatgcttc aataatattg aaaaaggaag agtcctgagg cggaaagaac
     3541 cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     3601 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     3661 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     3721 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     3781 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag
     3841 aagtagtgag gaggcttttt tggaggccta ggcttttgca aagatcgatc aagagacagg
     3901 atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg
     3961 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc
     4021 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg
     4081 tgccctgaat gaactgcaag acgaggcagc gcggctatcg tggctggcca cgacgggcgt
     4141 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg
     4201 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat
     4261 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca
     4321 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca
     4381 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa
     4441 ggcgagcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa
     4501 tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc
     4561 ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga
     4621 atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc
     4681 cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac
     4741 caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg
     4801 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc
     4861 atgctggagt tcttcgccca ccctaggggg aggctaactg aaacacggaa ggagacaata
     4921 ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca cggtgttggg
     4981 tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg
     5041 agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt
     5101 tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcctca
     5161 ggttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     5221 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     5281 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     5341 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     5401 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     5461 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     5521 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     5581 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     5641 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     5701 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     5761 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     5821 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     5881 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     5941 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     6001 aaccgtatta ccgccatgca ttagttatta atagtaatca attacggggt cattagttca
     6061 tagcccatat atggagttcc g
//
