pMpGE_En01 vector (V004675)

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LOCUS       40924_31270        6126 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Gateway entry vector pMpGE_En01 DNA, complete sequence.
ACCESSION   .
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  JOURNAL   Unpublished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)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6126)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (14-OCT-2015) Contact:Shigeo S Sugano Tokushima University, Center 
            for Collaboration among Agriculture, Industry, and Commerse"; 
            volume: " Kuramoto cho 3-18-15, Tokushima, Tokushima 770-8503, Japan
            URL :http"; pages: "//www.tokushima-u.ac.jp/ccaic"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6126
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     terminator      complement(387..473)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    569..668
                     /label=attL1
                     /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"
     promoter        2722..2752
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             2806..3483
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQAGRNLE
                     DPAY"
     CDS             3806..4108
                     /codon_start=1
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VPRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     misc_feature    4144..4148
                     /label=PstI
                     /note="PstI"
     misc_RNA        4149..4224
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    complement(4251..4350)
                     /label=attL2
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(4368..4386)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4391..4407)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             4520..5326
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /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"