LOCUS       Exported                6126 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Gateway entry vector pMpGE_En01 DNA, complete sequence.
ACCESSION   LC090754
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6126)
  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  JOURNAL   Unpublished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 6126)
  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 6126)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6126
                     /organism="Gateway entry vector pMpGE_En01"
                     /mol_type="other DNA"
                     /db_xref="taxon:1740675"
     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    689..2699
                     /label=MpU6-1pro
                     /note="MpU6-1pro"
     misc_feature    2700..2705
                     /label=SacI
                     /note="SacI"
     misc_feature    2706..4143
                     /label=CmR ccdB
                     /note="CmR ccdB"
     promoter        2722..2752
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             2806..3486
                     /codon_start=1
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQAGRNLE
                     DPAY"
     CDS             3806..4111
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VPRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     misc_feature    4144..4148
                     /label=PstI
                     /note="PstI"
     misc_feature    4149..4231
                     /label=sgRNA backbone
                     /note="sgRNA backbone"
     misc_RNA        4149..4224
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    4251..4350
                     /label=attL2
                     /note="attL2"
     protein_bind    complement(4251..4350)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(4367..4388)
                     /label=T7
                     /note="T7"
     promoter        complement(4368..4386)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4387..4407)
                     /label=M13-rev
                     /note="M13-rev"
     primer_bind     complement(4391..4407)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             4520..5329
                     /codon_start=1
                     /note="unnamed protein product; KanR"
                     /protein_id="BAT22813.1"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     rep_origin      5476..6064
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"