LOCUS       sequence-398032-        4797 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses a nuclear-targeted eGFP on a CMV promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4797)
  AUTHORS   Belin BJ, Cimini BA, Blackburn EH, Mullins RD
  TITLE     Visualization of actin filaments and monomers in somatic cell 
            nuclei.
  JOURNAL   Mol Biol Cell. 2013 Apr;24(7):982-94. doi: 10.1091/mbc.E12-09-0685. 
            Epub 2013 Feb 27.
  PUBMED    23447706
REFERENCE   2  (bases 1 to 4797)
  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: 
            "10.1091/mbc.E12-09-0685"; journalName: "Mol Biol Cell"; date: 
            "2013-04"; volume: "24"; issue: "7"; pages: "982-94"
COMMENT     Sequence Label: pEGFP-C1 EGFP-3XNLS
FEATURES             Location/Qualifiers
     source          1..4797
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        136..327
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     284..304
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(329..433)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      460..915
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(547..566)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     757..778
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    922..1043
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(933..952)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     987..1006
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(1218..1238)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(1242..1262)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(join(1299..2009,2010..2012,2013..2015))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(join(1299..2009,2010..2012,2013..2015))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1341..1362)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1690..1709
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1949..1970
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      2012..2021
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(2060..2263)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2089..2109)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2264..2567
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(2742..3330)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2822..2841)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3659..3706
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     3731..3750
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(3938..4732)
                     /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             complement(3938..4732)
                     /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(4049..4068)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4659..4678
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(join(4767..4797,1..327))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(4781..4797,1..119)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 cctcactact tctggaatag ctcagaggcc gaggcggcct cggcctctgc ataaataaaa
       61 aaaattagtc agccatgggg cggagaatgg gcggaactgg gcggagttag gggcgggatg
      121 ggcggagtta ggggcgggac tatggttgct gactaattga gatgcatgct ttgcatactt
      181 ctgcctgctg gggagcctgg ggactttcca cacctggttg ctgactaatt gagatgcatg
      241 ctttgcatac ttctgcctgc tggggagcct ggggactttc cacaccctaa ctgacacaca
      301 ttccacagct ggttctttcc gcctcaggac tcttcctttt tcaatattat tgaagcattt
      361 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
      421 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgcgccctgt agcggcgcat
      481 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
      541 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
      601 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
      661 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
      721 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
      781 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
      841 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
      901 taacgcttac aatttacgcg ttaagataca ttgatgagtt tggacaaacc acaactagaa
      961 tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca
     1021 ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc
     1081 agggggaggt gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt ggtatggctg
     1141 attatgatca gttatctaga tccggtggat cccgggcccg cggtacccta ttatcacttt
     1201 ctcttctttt ttggatctac ctttctcttc ttttttggat ctacctttct cttctttttt
     1261 ggatccatgg aagcttgagc tcgagatctg agtccggact tgtacagctc gtccatgccg
     1321 agagtgatcc cggcggcggt cacgaactcc agcaggacca tgtgatcgcg cttctcgttg
     1381 gggtctttgc tcagggcgga ctgggtgctc aggtagtggt tgtcgggcag cagcacgggg
     1441 ccgtcgccga tgggggtgtt ctgctggtag tggtcggcga gctgcacgct gccgtcctcg
     1501 atgttgtggc ggatcttgaa gttcaccttg atgccgttct tctgcttgtc ggccatgata
     1561 tagacgttgt ggctgttgta gttgtactcc agcttgtgcc ccaggatgtt gccgtcctcc
     1621 ttgaagtcga tgcccttcag ctcgatgcgg ttcaccaggg tgtcgccctc gaacttcacc
     1681 tcggcgcggg tcttgtagtt gccgtcgtcc ttgaagaaga tggtgcgctc ctggacgtag
     1741 ccttcgggca tggcggactt gaagaagtcg tgctgcttca tgtggtcggg gtagcggctg
     1801 aagcactgca cgccgtaggt cagggtggtc acgagggtgg gccagggcac gggcagcttg
     1861 ccggtggtgc agatgaactt cagggtcagc ttgccgtagg tggcatcgcc ctcgccctcg
     1921 ccggacacgc tgaacttgtg gccgtttacg tcgccgtcca gctcgaccag gatgggcacc
     1981 accccggtga acagctcctc gcccttgctc accatggtgg cgaccggtag cgctagcgga
     2041 tctgacggtt cactaaacca gctctgctta tatagacctc ccaccgtaca cgcctaccgc
     2101 ccatttgcgt caatggggcg gagttgttac gacattttgg aaagtcccgt tgattttggt
     2161 gccaaaacaa actcccattg acgtcaatgg ggtggagact tggaaatccc cgtgagtcaa
     2221 accgctatcc acgcccattg atgtactgcc aaaaccgcat caccatggta atagcgatga
     2281 ctaatacgta gatgtactgc caagtaggaa agtcccataa ggtcatgtac tgggcataat
     2341 gccaggcggg ccatttaccg tcattgacgt caataggggg cgtacttggc atatgataca
     2401 cttgatgtac tgccaagtgg gcagtttacc gtaaatactc cacccattga cgtcaatgga
     2461 aagtccctat tggcgttact atgggaacat acgtcattat tgacgtcaat gggcgggggt
     2521 cgttgggcgg tcagccaggc gggccattta ccgtaagtta tgtaacgcgg aactccatat
     2581 atgggctatg aactaatgac cccgtaattg attactatta ataactaatg catggcggta
     2641 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     2701 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     2761 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     2821 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     2881 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     2941 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     3001 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     3061 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     3121 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     3181 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     3241 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     3301 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     3361 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     3421 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     3481 gagtaacctg aggctatggc agggcctgcc gccccgacgt tggctgcgag ccctgggcct
     3541 tcacccgaac ttggggggtg gggtggggaa aaggaagaaa cgcgggcgta ttggccccaa
     3601 tggggtctcg gtggggtatc gacagagtgc cagccctggg accgaacccc gcgtttatga
     3661 acaaacgacc caacaccgtg cgttttattc tgtcttttta ttgccgtcat agcgcgggtt
     3721 ccttccggta ttgtctcctt ccgtgtttca gttagcctcc ccctagggtg ggcgaagaac
     3781 tccagcatga gatccccgcg ctggaggatc atccagccgg cgtcccggaa aacgattccg
     3841 aagcccaacc tttcatagaa ggcggcggtg gaatcgaaat ctcgtgatgg caggttgggc
     3901 gtcgcttggt cggtcatttc gaaccccaga gtcccgctca gaagaactcg tcaagaaggc
     3961 gatagaaggc gatgcgctgc gaatcgggag cggcgatacc gtaaagcacg aggaagcggt
     4021 cagcccattc gccgccaagc tcttcagcaa tatcacgggt agccaacgct atgtcctgat
     4081 agcggtccgc cacacccagc cggccacagt cgatgaatcc agaaaagcgg ccattttcca
     4141 ccatgatatt cggcaagcag gcatcgccat gggtcacgac gagatcctcg ccgtcgggca
     4201 tgctcgcctt gagcctggcg aacagttcgg ctggcgcgag cccctgatgc tcttcgtcca
     4261 gatcatcctg atcgacaaga ccggcttcca tccgagtacg tgctcgctcg atgcgatgtt
     4321 tcgcttggtg gtcgaatggg caggtagccg gatcaagcgt atgcagccgc cgcattgcat
     4381 cagccatgat ggatactttc tcggcaggag caaggtgaga tgacaggaga tcctgccccg
     4441 gcacttcgcc caatagcagc cagtcccttc ccgcttcagt gacaacgtcg agcacagctg
     4501 cgcaaggaac gcccgtcgtg gccagccacg atagccgcgc tgcctcgtct tgcagttcat
     4561 tcagggcacc ggacaggtcg gtcttgacaa aaagaaccgg gcgcccctgc gctgacagcc
     4621 ggaacacggc ggcatcagag cagccgattg tctgttgtgc ccagtcatag ccgaatagcc
     4681 tctccaccca agcggccgga gaacctgcgt gcaatccatc ttgttcaatc atgcgaaacg
     4741 atcctcatcc tgtctcttga tcgatctttg caaaagccta ggcctccaaa aaagcct
//
