LOCUS       sequence-90380-e        5478 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses a cytoplasmic actin filament reporter, the tandem CH 
            domains (a.a. 1-261) from human utrophin, on a CMV promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5478)
  AUTHORS   Belin BJ, Goins LM, Mullins RD
  TITLE     Comparative analysis of tools for live cell imaging of actin network
            architecture.
  JOURNAL   Bioarchitecture. 2015 Aug 28:1-14.
  PUBMED    26317264
REFERENCE   2  (bases 1 to 5478)
  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: 
            "Bioarchitecture. 2015 Aug 28:1-14."
COMMENT     Sequence Label: pEGFP-C1 Utr261-EGFP
FEATURES             Location/Qualifiers
     source          1..5478
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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      1389..1398
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1395..1397,1398..1400,1401..2111)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1395..1397,1398..1400,1401..2111)
                     /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(1440..1461)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1701..1720)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2048..2069
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    2112..2177
                     /label=MCS
                     /note="multiple cloning site"
     polyA_signal    2266..2387
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2303..2322)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2357..2376
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2394..2849)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2531..2552)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2743..2762
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2876..2980
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2982..3339
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2982..3173
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3005..3025)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3190..3325
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3374..4168
                     /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             3374..4168
                     /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(3428..3447)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4038..4057
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4356..4375)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4400..4447
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4776..5364
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5265..5284
                     /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 gatcctgaag atgttgccgt tcagcttcct gacaagaaat
     1321 ccataattat gtatttaaca tctttgtttg aggtgctacc tcagcaagtc accatagacg
     1381 taccggtcgc caccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc
     1441 tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg
     1501 gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg
     1561 tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc
     1621 ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg
     1681 agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg
     1741 agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca
     1801 acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg
     1861 acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca
     1921 gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc
     1981 tgcccgacaa ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc
     2041 gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg
     2101 agctgtacaa gtccggactc agatctcgag ctcaagcttc gaattctgca gtcgacggta
     2161 ccgcgggccc gggatccacc ggatctagat aatttacttg ctttaaaaaa cctcccacac
     2221 ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca
     2281 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     2341 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttaacg cgtaaattgt
     2401 aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa
     2461 ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt
     2521 gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa
     2581 agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag
     2641 ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt
     2701 tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg
     2761 agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc
     2821 cgcgcttaat gcgccgctac agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg
     2881 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     2941 accctgataa atgcttcaat aatattgaaa aaggaagagt cctgaggcgg aaagaaccag
     3001 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     3061 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     3121 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     3181 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     3241 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag
     3301 tagtgaggag gcttttttgg aggcctaggc ttttgcaaag atcgatcaag agacaggatg
     3361 aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt
     3421 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
     3481 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
     3541 cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
     3601 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
     3661 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat
     3721 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
     3781 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
     3841 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
     3901 gagcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat
     3961 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga
     4021 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg
     4081 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
     4141 ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa
     4201 gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg
     4261 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
     4321 ctggagttct tcgcccaccc tagggggagg ctaactgaaa cacggaagga gacaataccg
     4381 gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg tgttgggtcg
     4441 tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga
     4501 ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg
     4561 ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat agcctcaggt
     4621 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     4681 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     4741 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     4801 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     4861 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     4921 gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     4981 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     5041 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     5101 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     5161 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     5221 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     5281 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     5341 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     5401 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     5461 cgtattaccg ccatgcat
//
