pKW5_MB1Kan_TracrK_Spacer vector (V015883)

LOCUS       62056_13615        3180 bp DNA     circular SYN 30-AUG-2019
DEFINITION  Cloning vector pKW5_MB1Kan_TracrK_Spacer, complete sequence.
ACCESSION   MN226642
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3180)
  AUTHORS   Wang K, de la Torre D, Robertson WE, Chin JW.
  TITLE     Programmed chromosome fission and fusion enable precise large-scale 
            genome rearrangement and assembly
  JOURNAL   Science 365 (6456), 922-926 (2019)
REFERENCE   2  (bases 1 to 3180)
  AUTHORS   Wang K, de la Torre D, Robertson WE, Chin JW.
  TITLE     Direct Submission
  JOURNAL   Submitted (24-JUL-2019) Protein and Nucleic Acid Chemistry, MRC 
            Laboratory of Molecular Biology, Francis Crick Avenue, Cambridge, 
            Cambridgeshire CB2 0QH, United Kingdom
REFERENCE   3  (bases 1 to 3180)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Science"; 
            date: "2019"; volume: "365"; issue: "6456"; pages: "922-926"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (24-JUL-2019) Protein and Nucleic Acid Chemistry, MRC Laboratory of 
            Molecular Biology, Francis Crick Avenue, Cambridge, Cambridgeshire 
            CB2 0QH, United Kingdom"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..3180
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        104..208
                     /label=AmpR promoter
     CDS             209..1000
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     rep_origin      1174..1762
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(1926..2012)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     regulatory      2168..2202
                     /label=putative Cas9 promoter
                     /note="putative Cas9 promoter"
                     /regulatory_class="promoter"
     regulatory      2234..2261
                     /label=putative tracrRNA promoter
                     /note="putative tracrRNA promoter"
                     /regulatory_class="promoter"
     misc_RNA        2315..2393
                     /label=tracrRNA
                     /note="trans-activating CRISPR RNA for the Streptococcus 
                     pyogenes CRISPR/Cas9 system"
     misc_feature    2505..2636
                     /label=crRNA leader
                     /note="crRNA leader sequence for the Streptococcus pyogenes
                     CRISPR/Cas system"
     repeat_region   2637..2672
                     /label=DR
                     /note="direct repeat for the Streptococcus pyogenes
                     CRISPR/Cas system"
     gap             2673..2702
                     /estimated_length=30
     gap             2739..2768
                     /estimated_length=30
     gap             2805..2834
                     /estimated_length=30
     gap             2871..2900
                     /estimated_length=30
     gap             2937..2966
                     /estimated_length=30
     gap             3003..3032
                     /estimated_length=30
     promoter        complement(3101..3174)
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator
                     sites (Lutz and Bujard, 1997)"