LOCUS       Exported                5498 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Expresses a nuclear-targeted actin filament reporter, consisting of 
            the first 230 amino acids of human utrophin, on a CMV promoter.
ACCESSION   .
VERSION     .
KEYWORDS    pEGFP-C1 Utr230-EGFP-3XNLS
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5498)
  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 5498)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5498
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1308..1317
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1314..2030
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             1314..2030
                     /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(1359..1380)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1620..1639)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1967..1988
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2067..2087
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2091..2111
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    2286..2407
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2323..2342)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2377..2396
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2414..2869)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2551..2572)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2763..2782
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2896..3000
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3002..3359
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(3025..3045)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3210..3345
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3272..3291
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3394..4188
                     /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(3448..3467)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4058..4077
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4376..4395)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4420..4467
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4796..5384
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5285..5304
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcatgg
      601 ccaagtatgg agaacatgaa gccagtcctg acaatgggca gaacgaattc agtgatatca
      661 ttaagtccag atctgatgaa cacaatgacg tacagaagaa aacctttacc aaatggataa
      721 atgctcgatt ttcaaagagt gggaaaccac ccatcaatga tatgttcaca gacctcaaag
      781 atggaaggaa gctattggat cttctagaag gcctcacagg aacatcactg ccaaaggaac
      841 gtggttccac aagggtacat gccttaaata acgtcaacag agtgctgcag gttttacatc
      901 agaacaatgt ggaattagtg aatatagggg gaactgacat tgtggatgga aatcacaaac
      961 tgactttggg gttactttgg agcatcattt tgcactggca ggtgaaagat gtcatgaagg
     1021 atgtcatgtc ggacctgcag cagacgaaca gtgagaagat cctgctcagc tgggtgcgtc
     1081 agaccaccag gccctacagc caagtcaacg tcctcaactt caccaccagc tggacagatg
     1141 gactcgcctt taatgctgtc ctccaccgac ataaacctga tctcttcagc tgggataaag
     1201 ttgtcaaaat gtcaccaatt gagagacttg aacatgcctt cagcaaggct caaacttatt
     1261 tgggaattga aaagctgtta gatcctgggg gggggggggt accggtcgcc accatggtga
     1321 gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg
     1381 taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc
     1441 tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga
     1501 ccaccctgac ctacggcgtg cagtgcttca gccgctaccc cgaccacatg aagcagcacg
     1561 acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg
     1621 acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc
     1681 gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg
     1741 agtacaacta caacagccac aacgtctata tcatggccga caagcagaag aacggcatca
     1801 aggtgaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc gccgaccact
     1861 accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga
     1921 gcacccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg
     1981 agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag tccggactca
     2041 gatctcgagc tcaagcttcc atggatccaa aaaagaagag aaaggtagat ccaaaaaaga
     2101 agagaaaggt agatccaaaa aagaagagaa agtgataata gggtaccgcg ggcccgggat
     2161 ccaccggatc tagataactg atcataatca gccataccac atttgtagag gttttacttg
     2221 ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg
     2281 ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     2341 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     2401 tatcttaacg cgtaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt
     2461 taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa
     2521 gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag
     2581 aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt
     2641 gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac
     2701 cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag
     2761 gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg
     2821 cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc aggtggcact
     2881 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     2941 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     3001 cctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
     3061 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     3121 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     3181 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     3241 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg
     3301 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaag
     3361 atcgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca
     3421 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
     3481 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
     3541 aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg
     3601 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
     3661 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
     3721 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
     3781 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
     3841 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
     3901 ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt gacccatggc
     3961 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
     4021 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
     4081 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
     4141 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg
     4201 ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg
     4261 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc
     4321 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc tagggggagg ctaactgaaa
     4381 cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa agacagaata
     4441 aaacgcacgg tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact
     4501 ctgtcgatac cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc
     4561 caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag
     4621 gccctgccat agcctcaggt tactcatata tactttagat tgatttaaaa cttcattttt
     4681 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     4741 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     4801 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     4861 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     4921 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     4981 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     5041 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     5101 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     5161 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     5221 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     5281 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     5341 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     5401 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     5461 cccctgattc tgtggataac cgtattaccg ccatgcat
//
