LOCUS       sequence-265898-        5829 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes YFP and NLS tagged beta-actin with the G13R depolymerization
            mutation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5829)
  AUTHORS   Chang L, Godinez WJ, Kim IH, Tektonidis M, de Lanerolle P, Eils R, 
            Rohr K, Knipe DM
  TITLE     Herpesviral replication compartments move and coalesce at nuclear 
            speckles to enhance export of viral late mRNA.
  JOURNAL   Proc Natl Acad Sci U S A. 2011 May 24;108(21):E136-44. doi: 
            10.1073/pnas.1103411108. Epub 2011 May 9.
  PUBMED    21555562
REFERENCE   2  (bases 1 to 5829)
  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"; doi: 
            "10.1073/pnas.1103411108"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2011-05-24- 24"; volume: "108"; issue: "21"; pages: "E136-44"
COMMENT     Sequence Label: YFP NLS Beta-Actin G13R
FEATURES             Location/Qualifiers
     source          1..5829
                     /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="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDLKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(598..619)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     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"
     CDS             1270..1290
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    2557..2678
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2594..2613)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2648..2667
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2685..3140)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2822..2843)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3034..3053
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3167..3271
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3273..3630
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3273..3464
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3296..3316)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3481..3616
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3665..4459
                     /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             3665..4459
                     /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(3719..3738)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4329..4348
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4647..4666)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4691..4738
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5067..5655
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5556..5575
                     /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 ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
      601 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
      661 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
      721 ccctggccca ccctcgtgac caccttcggc tacggcctgc agtgcttcgc ccgctacccc
      781 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
      841 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
      901 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acctcaagga ggacggcaac
      961 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     1021 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     1081 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1141 cccgacaacc actacctgag ctaccagtcc gccctgagca aagaccccaa cgagaagcgc
     1201 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     1261 ctgtacaagc caaaaaagaa gagaaaggta cctcgagcta tggatgatga tatcgccgcg
     1321 ctcgtcgtcg acaaccgctc cggcatgtgc aaggccggct tcgcgggcga cgatgccccc
     1381 cgggccgtct tcccctccat cgtggggcgc cccaggcacc agggcgtgat ggtgggcatg
     1441 ggtcagaagg attcctatgt gggcgacgag gcccagagca agagaggcat cctcaccctg
     1501 aagtacccca tcgagcacgg catcgtcacc aactgggacg acatggagaa aatctggcac
     1561 cacaccttct acaatgagct gcgtgtggct cccgaggagc accccgtgct gctgaccgag
     1621 gcccccctga accccaaggc caaccgcgag aagatgaccc agatcatgtt tgagaccttc
     1681 aacaccccag ccatgtacgt tgctatccag gctgtgctat ccctgtacgc ctctggccgt
     1741 accactggca tcgtgatgga ctccggtgac ggggtcaccc acactgtgcc catctacgag
     1801 gggtatgccc tcccccatgc catcctgcgt ctggacctgg ctggccggga cctgactgac
     1861 tacctcatga agatcctcac cgagcgcggc tacagcttca ccaccacggc cgagcgggaa
     1921 atcgtgcgtg acattaagga gaagctgtgc tacgtcgccc tggacttcga gcaagagatg
     1981 gccacggctg cttccagctc ctccctggag aagagctacg agctgcctga cggccaggtc
     2041 atcaccattg gcaatgagcg gttccgctgc cctgaggcac tcttccagcc ttccttcctg
     2101 ggcatggagt cctgtggcat ccacgaaact accttcaact ccatcatgaa gtgtgacgtg
     2161 gacatccgca aagacctgta cgccaacaca gtgctgtctg gcggcaccac catgtaccct
     2221 ggcattgccg acaggatgca gaaggagatc actgccctgg cacccagcac aatgaagatc
     2281 aagatcattg ctcctcctga gcgcaagtac tccgtgtgga tcggcggctc catcctggcc
     2341 tcgctgtcca ccttccagca gatgtggatc agcaagcagg agtatgacga gtccggcccc
     2401 tccatcgtcc accgcaaatg cttctaggga tccaccggat ctagataact gatcataatc
     2461 agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg
     2521 aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat
     2581 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     2641 tctagttgtg gtttgtccaa actcatcaat gtatcttaag gcgtaaattg taagcgttaa
     2701 tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc
     2761 cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt
     2821 tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa
     2881 aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg
     2941 gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg
     3001 acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc
     3061 tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa
     3121 tgcgccgcta cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     3181 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     3241 aatgcttcaa taatattgaa aaaggaagag tcctgaggcg gaaagaacca gctgtggaat
     3301 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     3361 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     3421 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     3481 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     3541 tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga
     3601 ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt
     3661 tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct
     3721 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct
     3781 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga
     3841 actgcaagac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc
     3901 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg
     3961 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc
     4021 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca
     4081 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga
     4141 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgagcatgcc
     4201 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga
     4261 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca
     4321 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg
     4381 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct
     4441 tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc
     4501 aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
     4561 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
     4621 ttcgcccacc ctagggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc
     4681 cgcgctatga cggcaataaa aagacagaat aaaacgcacg gtgttgggtc gtttgttcat
     4741 aaacgcgggg ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg
     4801 ggccaatacg cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc
     4861 ccagggctcg cagccaacgt cggggcggca ggccctgcca tagcctcagg ttactcatat
     4921 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     4981 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     5041 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     5101 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     5161 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     5221 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     5281 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     5341 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     5401 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     5461 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     5521 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     5581 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     5641 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     5701 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     5761 gccatgcatt agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat
     5821 ggagttccg
//
