pKill3 vector (V015625)

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LOCUS       62056_13395        8722 bp DNA     circular SYN 07-JUL-2019
DEFINITION  Cloning vector pKill3, complete sequence.
ACCESSION   MK756313
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8722)
  AUTHORS   Neil K, Allard N, Jordan D, Rodrigue S.
  TITLE     Assembly of large mobilizable genetic cargo by double recombinase 
            operated insertion of DNA (DROID)
  JOURNAL   Plasmid (2019) In press
  PUBMED    31247227
REFERENCE   2  (bases 1 to 8722)
  AUTHORS   Neil K, Rodrigue S.
  TITLE     Direct Submission
  JOURNAL   Submitted (04-APR-2019) Biology, University of Sherbrooke, 2500 
            Boulevard de l'Universite, Sherbrooke, Quebec J1K 2R1, Canada
REFERENCE   3  (bases 1 to 8722)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Plasmid 
            (2019) In press"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (04-APR-2019) Biology, University of Sherbrooke, 2500 Boulevard de 
            l'Universite, Sherbrooke, Quebec J1K 2R1, Canada"
COMMENT     ##Assembly-Data-START##
            Assembly Method       :: ApE v. 2018-05-04
            Sequencing Technology :: Putative Sequence
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..8722
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1..58
                     /label=attPbxb1
                     /note="attPbxb1"
     rep_origin      539..761
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             809..1756
                     /label=Rep101(Ts)
                     /note="temperature-sensitive version of the RepA protein
                     needed for replication with the pSC101 origin (Armstrong et
                     al., 1984)"
     protein_bind    2192..2239
                     /label=FRT
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     regulatory      2267..2348
                     /label=apFAB391
                     /note="apFAB391"
                     /regulatory_class="terminator"
     regulatory      2349..2398
                     /allele="PN25"
                     /regulatory_class="promoter"
     misc_feature    2399..2418
                     /label=spacer
                     /note="spacer"
     misc_RNA        2419..2494
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     regulatory      2499..2529
                     /label=B1006U10
                     /note="B1006U10"
                     /regulatory_class="terminator"
     regulatory      2590..2612
                     /label=apFAB391
                     /note="apFAB391"
                     /regulatory_class="terminator"
     regulatory      2613..2662
                     /label=PN25
                     /note="PN25"
                     /regulatory_class="promoter"
     misc_RNA        2663..2762
                     /product="gRNA_2"
     misc_feature    2663..2682
                     /label=spacer
                     /note="spacer"
     regulatory      2763..2793
                     /label=B1006U10
                     /note="B1006U10"
                     /regulatory_class="terminator"
     regulatory      2854..2876
                     /label=apFAB391
                     /note="apFAB391"
                     /regulatory_class="terminator"
     regulatory      2877..2926
                     /allele="PN25"
                     /regulatory_class="promoter"
     misc_RNA        2927..3026
                     /product="gRNA_3"
     misc_feature    2927..2946
                     /label=spacer
                     /note="spacer"
     regulatory      3027..3057
                     /label=B1006U10
                     /note="B1006U10"
                     /regulatory_class="terminator"
     regulatory      3255..3304
                     /regulatory_class="promoter"
     CDS             3393..4184
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     CDS             4616..8719
                     /label=Cas9
                     /note="Cas9 (Csn1) endonuclease from the Streptococcus
                     pyogenes Type II CRISPR/Cas system"