LOCUS       Exported                3214 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Gateway entry vector pMpGE_En03 DNA, complete sequence.
ACCESSION   LC090755
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3214)
  AUTHORS   Sugano SS, Nishihama R, Shirakawa M, Matsuda Y, Takagi T, 
            Hara-Nishimura I, Osakabe K, Kohchi T.
  TITLE     Efficient CRISPR/Cas9 vectors in Marchantia polymorpha
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 3214)
  AUTHORS   Sugano SS, Kohchi T, Nishihama R.
  TITLE     Direct Submission
  JOURNAL   Submitted (14-OCT-2015) Contact:Shigeo S Sugano Tokushima 
            University, Center for Collaboration among Agriculture, Industry, 
            and Commerse; Kuramoto cho 3-18-15, Tokushima, Tokushima 770-8503, 
            Japan URL :http://www.tokushima-u.ac.jp/ccaic/
REFERENCE   3  (bases 1 to 3214)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..3214
                     /organism="Gateway entry vector pMpGE_En03"
                     /mol_type="other DNA"
                     /db_xref="taxon:1740676"
     terminator      268..295
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      387..473
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     536..553
                     /label=M13-fwd
                     /note="M13-fwd"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    569..668
                     /label=attL1
                     /note="attL1"
     protein_bind    569..668
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     misc_feature    685..1199
                     /label=MpU6-1 promoter (500 bp)
                     /note="MpU6-1 promoter (500 bp)"
     misc_feature    1200
                     /label=initial G
                     /note="initial G"
     misc_feature    complement(1202..1207)
                     /label=BsaI
                     /note="BsaI"
     misc_feature    1215..1220
                     /label=BsaI
                     /note="BsaI"
     misc_feature    1222..1304
                     /label=sgRNA backbone
                     /note="sgRNA backbone"
     misc_RNA        1222..1297
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    1339..1438
                     /label=attL2
                     /note="attL2"
     protein_bind    complement(1339..1438)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1455..1476)
                     /label=T7
                     /note="T7"
     promoter        complement(1456..1474)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1475..1495)
                     /label=M13-rev
                     /note="M13-rev"
     primer_bind     complement(1479..1495)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             1608..2417
                     /codon_start=1
                     /note="unnamed protein product; KanR"
                     /protein_id="BAT22814.1"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     rep_origin      2564..3152
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"