pVMG0218_Cq-U6-4_2xBbsI-gRNA vector (V016513)

LOCUS       62056_22625        5789 bp DNA     circular SYN 26-APR-2021
DEFINITION  Cloning vector pVMG0218_Cq-U6-4_2xBbsI-gRNA, complete sequence.
ACCESSION   MW925702
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5789)
  AUTHORS   Feng X, Lopez Del Amo V, Mameli E, Lee M, Bishop AL, Perrimon N, 
            Gantz VM.
  TITLE     Optimized CRISPR tools and site-directed transgenesis towards gene 
            drive development in Culex quinquefasciatus mosquitoes
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5789)
  AUTHORS   Feng X, Lopez Del Amo V, Mameli E, Lee M, Bishop AL, Perrimon N, 
            Gantz VM.
  TITLE     Direct Submission
  JOURNAL   Submitted (05-APR-2021) Biology Sciences, University of California, 
            San Diego, 9500 Gilman Dr., La Jolla, CA 92093, USA
REFERENCE   3  (bases 1 to 5789)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (05-APR-2021) Biology Sciences, University of California, San Diego,
            9500 Gilman Dr., La Jolla, CA 92093, USA"
FEATURES             Location/Qualifiers
     source          1..5789
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..429
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     CDS             773..1564
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     CDS             1585..2442
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      2616..3204
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    3492..3513
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        3528..3558
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3566..3582
                     /label=lac operator
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     3590..3606
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     3'UTR           complement(3680..4188)
                     /note="3'UTR of Culex quinquefasciatus U6-4 gene
                     (CPIJ039801)"
     misc_RNA        complement(4189..4264)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    complement(4265..4284)
                     /note="2xBsmBI spacer; gRNA insertion site"
     5'UTR           complement(4285..5537)
                     /note="regulatory region of Culex quinquefasciatus U6-4
                     gene (CPIJ039801)"
     promoter        complement(5608..5626)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5633..5649)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"