pCasRx-3 vector (V015685)

LOCUS       62056_4920        7902 bp DNA     circular SYN 09-AUG-2020
DEFINITION  Cloning vector pCasRx-3, complete sequence.
ACCESSION   MN934322
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7902)
  AUTHORS   Zhang K, Zhang Z, Kang J, Chen J, Liu J, Gao N, Fan L, Zheng P, Wang
            Y, Sun J.
  TITLE     CRISPR/Cas13d-Mediated Microbial RNA Knockdown
  JOURNAL   Front Bioeng Biotechnol 8, 856 (2020)
REFERENCE   2  (bases 1 to 7902)
  AUTHORS   Wang Y.
  TITLE     Direct Submission
  JOURNAL   Submitted (13-JAN-2020) Key Laboratory of Systems Microbial 
            Biotechnology, Tianjin Institute of Industrial Biotechnology, 
            Chinese Academy of Sciences, 32 West 7th Avenue, Tianjin Airport 
            Economic Area, Tianjin 300308, China
REFERENCE   3  (bases 1 to 7902)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Front 
            Bioeng Biotechnol 8, 856 (2020)"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (13-JAN-2020) Key Laboratory of Systems Microbial Biotechnology, 
            Tianjin Institute of Industrial Biotechnology, Chinese Academy of 
            Sciences, 32 West 7th Avenue, Tianjin Airport Economic Area, Tianjin
            300308, China"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..7902
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..19
                     /label=tet operator
                     /note="bacterial operator O2 for the tetR and tetA genes"
     regulatory      55..67
                     /regulatory_class="ribosome_binding_site"
     CDS             68..2968
                     /label=RfxCas13d
                     /note="RNA-targeting Type VI CRISPR protein from
                     Ruminococcus flavefaciens strain XPD3002 (Konermann et al.,
                     2018)"
     terminator      3078..3105
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     promoter        complement(3274..3329)
                     /label=tetR/tetA promoters
                     /note="overlapping promoters for bacterial tetR and tetA"
     CDS             3345..3968
                     /label=TetR
                     /note="tetracycline repressor TetR"
     rep_origin      5086..5308
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             5356..6303
                     /label=Rep101
                     /note="RepA protein needed for replication with the pSC101 
                     origin"
     promoter        6518..6621
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     CDS             6622..7278
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
     terminator      7292..7351
                     /label=his operon terminator
                     /note="This putative transcriptin terminator from the E.
                     coli his operon has a 2-bp deletion introduced during 
                     synthesis. Its efficiency has not been determined."
     terminator      complement(7411..7440)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     primer_bind     complement(7506..7522)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(7559..7577)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(7598..7614)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(7634..7655)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."