LOCUS       Exported               12653 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  AAVS1 targeting vector pA1.GFP.A2, complete sequence.
ACCESSION   GQ380658
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12653)
  AUTHORS   van Nierop GP, de Vries AA, Holkers M, Vrijsen KR, Goncalves MA.
  TITLE     Stimulation of homology-directed gene targeting at an endogenous 
            human locus by a nicking endonuclease
  JOURNAL   Nucleic Acids Res. 37 (17), 5725-5736 (2009)
  PUBMED    19651880
REFERENCE   2  (bases 1 to 12653)
  AUTHORS   van Nierop GP, de Vries AA, Holkers M, Vrijsen KR, Goncalves MA.
  TITLE     Direct Submission
  JOURNAL   Submitted (14-JUL-2009) Molecular Cell Biology, Leiden University 
            Medical Center, Einthovenweg 20, Leiden, Zuid Holland 2333 ZC, The 
            Netherlands
REFERENCE   3  (bases 1 to 12653)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..12653
                     /organism="AAVS1 targeting vector pA1.GFP.A2"
                     /mol_type="other DNA"
                     /db_xref="taxon:672173"
     regulatory      1..2456
                     /regulatory_class="promoter"
                     /note="human elongation factor 1 alpha promoter"
     promoter        1216..2397
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          1446..2388
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     5UTR           2457..2855
                     /note="5untranslated region of SV40 early genes (S) and 
                     HTLV-1 (UR)"
     LTR             2505..2773
                     /label=5 LTR (truncated)
                     /note="truncated 5 long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     promoter        complement(2810..2828)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2895..3614
                     /codon_start=1
                     /gene="hrGFP"
                     /product="hrGFP"
                     /label=hrGFP
                     /note="green fluorescence protein"
                     /protein_id="ACV74423.1"
                     /translation="MVSKQILKNTGLQEIMSFKVNLEGVVNNHVFTMEGCGKGNILFGN
                     QLVQIRVTKGAPLPFAFDILSPAFQYGNRTFTKYPEDISDFFIQSFPAGFVYERTLRYE
                     DGGLVEIRSDINLIEEMFVYRVEYKGRNFPNDGPVMKKTITGLQPSFEVVYMNDGVLVG
                     QVILVYRLNSGKFYSCHMRTLMKSKGVVKDFPEYHFIQHRLEKTYVEDGGFVEQHETAI
                     AQLTSLGKPLGSLHEWV"
     gene            2895..3614
                     /gene="hrGFP"
                     /label=hrGFP
     regulatory      3637..3855
                     /regulatory_class="polyA_signal_sequence"
                     /note="SV40 poly A"
     polyA_signal    3646..3767
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_feature    4059..8438
                     /note="Arm 2; human chromosome 19 AAVS1 locus region"
     misc_feature    4062..4563
                     /label=HA-L
                     /note="left homology arm from the adeno-associated virus 
                     integration site (AAVS1) within intron 1 of the human 
                     PPP1R12C gene"
     misc_feature    4685..5521
                     /label=HA-R
                     /note="right homology arm from the adeno-associated virus 
                     integration site (AAVS1) within intron 1 of the human 
                     PPP1R12C gene"
     rep_origin      8629..8985
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     oriT            9035..9330
                     /label=Escherichia coli OriT
                     /note="Escherichia coli OriT"
     oriT            9035..9325
                     /note="origin of transfer for the bacterial F plasmid 
                     (Frost et al., 1994)"
     CDS             complement(9378..10172)
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     misc_feature    10588..12626
                     /note="Arm 1; human chromosome 19 AAVS1 locus region"