pCDF-Crispr-CasWT vector (V008638)

LOCUS       40924_9396        8519 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pCDF-Crispr-CasWT DNA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8519)
  AUTHORS   Hamamoto K, Tokunaga T, Yagi Y, Hirai I.
  TITLE     Characterization of blaCTX-M-14 transposition from plasmid to 
            chromosome in Escherichia coli model strain
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8519)
  AUTHORS   Hamamoto K, Hirai I.
  TITLE     Direct Submission
  JOURNAL   Submitted (22-MAR-2018) Contact:Kouta Hamamoto University of the 
            Ryukyus, Microbiology; 207 Uehara, Nishihara-cho, Nakagami-gun, 
            Okinawa 903-0215, Japan
REFERENCE   3  (bases 1 to 8519)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8519)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (22-MAR-2018) Contact:Kouta Hamamoto University of the Ryukyus, 
            Microbiology; 207 Uehara, Nishihara-cho, Nakagami-gun, Okinawa 
            903-0215, Japan"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8519
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(61..684)
                     /label=TetR
                     /note="tetracycline repressor TetR"
     promoter        700..755
                     /label=tetR/tetA promoters
                     /note="overlapping promoters for bacterial tetR and tetA"
     RBS             773..784
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             791..4894
                     /label=Cas9
                     /note="Cas9 (Csn1) endonuclease from the Streptococcus
                     pyogenes Type II CRISPR/Cas system"
     terminator      4921..4992
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      5015..5062
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(5236..6024)
                     /label=SmR
                     /note="aminoglycoside adenylyltransferase (Murphy, 1985)"
     promoter        complement(6025..6116)
                     /label=AmpR promoter
     rep_origin      complement(6164..6902)
                     /direction=LEFT
                     /label=CloDF13 ori
                     /note="Plasmids containing the CloDF13 (CDF) origin of 
                     replication can be propagated in E. coli cells that contain
                     additional plasmids with compatible origins."
     misc_RNA        complement(6939..7014)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    complement(7044..7060)
                     /label=lac operator
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(7068..7097)
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     CDS             complement(7332..8411)
                     /label=lacI
                     /note="lac repressor"
     promoter        complement(8412..8489)
                     /label=lacI promoter