LOCUS       Exported                4868 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Expresses a cytoplasmic actin filament reporter, the neuronal 
            inositol 1,4,5-triphosphate 3-kinase A actin-binding domain known as
            F-tractin, on a CMV promoter.
ACCESSION   .
VERSION     .
KEYWORDS    pEGFP-C1 F-tractin-EGFP
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4868)
  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 4868)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4868
                     /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      744..753
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             750..1466
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             750..1466
                     /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(795..816)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1056..1075)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1403..1424
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    1467..1532
                     /label=MCS
                     /note="multiple cloning site"
     polyA_signal    1656..1777
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1693..1712)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1747..1766
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1784..2239)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1921..1942)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2133..2152
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2266..2370
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2372..2729
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(2395..2415)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2580..2715
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2642..2661
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2764..3558
                     /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(2818..2837)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3428..3447
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3746..3765)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3790..3837
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4166..4754
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4655..4674
                     /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 cgcgaccacg gggcgcgggg ccctgcagcc ccgggttgga gcgggctccg cgccggagcg
      661 tcggggagct gcgcctgctc ttcgaagcgc gctgcgccgc agtcgccgcc gcagcggccg
      721 caggggggct agcgctaccg gtcgccacca tggtgagcaa gggcgaggag ctgttcaccg
      781 gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt
      841 ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca
      901 ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt
      961 gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg
     1021 aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg
     1081 ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact
     1141 tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg
     1201 tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca
     1261 acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg
     1321 acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag
     1381 accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca
     1441 ctctcggcat ggacgagctg tacaagtccg gactcagatc tcgagctcaa gcttcgaatt
     1501 ctgcagtcga cggtaccgcg ggcccgggat ccaccggatc tagataactg atcataatca
     1561 gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga
     1621 acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg
     1681 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     1741 ctagttgtgg tttgtccaaa ctcatcaatg tatcttaacg cgtaaattgt aagcgttaat
     1801 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc
     1861 gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt
     1921 ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa
     1981 accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg
     2041 tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga
     2101 cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct
     2161 agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat
     2221 gcgccgctac agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     2281 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     2341 atgcttcaat aatattgaaa aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg
     2401 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     2461 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     2521 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     2581 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     2641 ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag
     2701 gcttttttgg aggcctaggc ttttgcaaag atcgatcaag agacaggatg aggatcgttt
     2761 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     2821 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     2881 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     2941 ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     3001 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     3061 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     3121 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     3181 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     3241 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc
     3301 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     3361 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     3421 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     3481 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     3541 cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca
     3601 acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
     3661 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
     3721 tcgcccaccc tagggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc
     3781 gcgctatgac ggcaataaaa agacagaata aaacgcacgg tgttgggtcg tttgttcata
     3841 aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg
     3901 gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc
     3961 cagggctcgc agccaacgtc ggggcggcag gccctgccat agcctcaggt tactcatata
     4021 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     4081 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     4141 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     4201 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     4261 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     4321 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     4381 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     4441 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     4501 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     4561 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     4621 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     4681 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     4741 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     4801 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     4861 ccatgcat
//
