ds-spcas-vqr vector (V010005)

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LOCUS       40924_690        6806 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector ds-spcas-vqr, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6806)
  AUTHORS   Chavez A, Pruitt BW, Tuttle M, Shapiro RS, Cecchi RJ, Winston J, 
            Turczyk BM, Tung M, Collins JJ, Church GM.
  TITLE     Precise Cas9 targeting enables genomic mutation prevention
  JOURNAL   Proc. Natl. Acad. Sci. U.S.A. (2018) In press
  PUBMED    29555762
REFERENCE   2  (bases 1 to 6806)
  AUTHORS   Chavez A, Pruitt BW, Tuttle M, Shapiro RS, Cecchi RJ, Winston J, 
            Turczyk BM, Tung M, Collins JJ, Church GM.
  TITLE     Direct Submission
  JOURNAL   Submitted (02-MAR-2018) Synthetic Biology, Wyss Institute, 3 
            Blackfan Circle, Floor 5, Boston, MA 02115, USA
REFERENCE   3  (bases 1 to 6806)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6806)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Proc. Natl.
            Acad. Sci. U.S.A. (2018) In press"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (02-MAR-2018) Synthetic Biology, Wyss Institute, 3 Blackfan Circle, 
            Floor 5, Boston, MA 02115, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6806
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      153..180
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     regulatory      205..364
                     /label=proC promoter
                     /note="proC promoter"
                     /regulatory_class="promoter"
     CDS             377..4480
                     /label=Cas9 VQR
                     /note="Streptococcus pyogenes Cas9 endonuclease with the 
                     D1135V/R1335Q/T1337R mutations"
     misc_RNA        4551..4629
                     /label=tracrRNA
                     /note="trans-activating CRISPR RNA for the Streptococcus 
                     pyogenes CRISPR/Cas9 system"
     regulatory      complement(4667..4708)
                     /note="derived from Streptococcus pyogenes; tracrRNA
                     terminator"
                     /regulatory_class="terminator"
     repeat_region   4845..4880
                     /label=DR
                     /note="direct repeat for the Streptococcus pyogenes
                     CRISPR/Cas system"
     rep_origin      5062..5800
                     /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."
     promoter        5848..5939
                     /label=AmpR promoter
     CDS             5940..6728
                     /label=SmR
                     /note="aminoglycoside adenylyltransferase (Murphy, 1985)"