pKW20_CDFtet_pAraRedCas9_TracrRNA_rec vector (V015990)

LOCUS       62056_13605        9863 bp DNA     circular SYN 12-JUN-2021
DEFINITION  Cloning vector pKW20_CDFtet_pAraRedCas9_TracrRNA_rec, complete 
            sequence.
ACCESSION   MW879721
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9863)
  AUTHORS   Robertson WE, Funke LFH., de la Torre D, Fredens J, Elliott TS, 
            Spinck M, Christova Y, Cervettini D, Boge FL, Liu KC, Buse S, Maslen
            S, Salmond GPC., Chin JW.
  TITLE     Sense Codon Reassignment Enables Viral Resistance and Encoded 
            Polymer Synthesis
  JOURNAL   Science (2021) In press
REFERENCE   2  (bases 1 to 9863)
  AUTHORS   Robertson WE, Funke LFH., de la Torre D, Fredens J, Elliott TS, 
            Spinck M, Christova Y, Cervettini D, Boge FL, Liu KC, Buse S, Maslen
            S, Salmond GPC., Chin JW.
  TITLE     Direct Submission
  JOURNAL   Submitted (06-APR-2021) Protein and Nucleic Acid Chemistry, MRC 
            Laboratory of Molecular Biology, Francis Crick Avenue, Cambridge, 
            Cambridgeshire CB2 0QH, United Kingdom
REFERENCE   3  (bases 1 to 9863)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Science 
            (2021) In press"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (06-APR-2021) 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..9863
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_RNA        complement(106..184)
                     /label=tracrRNA
                     /note="trans-activating CRISPR RNA for the Streptococcus 
                     pyogenes CRISPR/Cas9 system"
     misc_feature    complement(238..265)
                     /label=tracrRNA promoter predicted
                     /note="tracrRNA promoter predicted"
     misc_feature    297..331
                     /label=Cas9 promoter predicted
                     /note="Cas9 promoter predicted"
     terminator      470..497
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     CDS             complement(670..1545)
                     /label=araC
                     /note="L-arabinose regulatory protein"
     promoter        1572..1856
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     CDS             1891..2304
                     /label=Gam
                     /note="inhibitor of the host RecBCD nuclease in the lambda 
                     Red system"
     CDS             2313..3095
                     /label=Beta
                     /note="single-stranded DNA binding recombinase in the 
                     lambda Red system"
     CDS             3095..3772
                     /label=Exo
                     /note="5' to 3' double-stranded DNA exonuclease in the 
                     lambda
                     Red system"
     misc_feature    3773..3775
                     /label=TAG to TAA
                     /note="TAG to TAA"
     CDS             3799..7902
                     /label=Cas9
                     /note="Cas9 (Csn1) endonuclease from the Streptococcus
                     pyogenes Type II CRISPR/Cas system"
     terminator      7942..7989
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(8163..8963)
                     /label=ApmR
                     /note="aminoglycoside 3-N-acetyltransferase type IV"
     promoter        complement(8964..9055)
                     /label=AmpR promoter
     rep_origin      complement(9103..9841)
                     /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."