pVMG0109_CC-U6-3-w5_GFP_wHAs_O vector (V015814)

LOCUS       62056_22600        9522 bp DNA     circular SYN 26-APR-2021
DEFINITION  Cloning vector pVMG0109_CC-U6-3-w5_GFP_wHAs_O, complete sequence.
ACCESSION   MW925706
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9522)
  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  JOURNAL   Unpublished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 9522)
  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 9522)
  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            "Unpublished"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..9522
                     /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"
     misc_feature    3680..4724
                     /label=white homology arm (left)
                     /note="white homology arm (left)"
     3'UTR           4746..6414
                     /label=3'UTR of Dmel actin5c (FBtr0345894)
                     /note="3'UTR of Dmel actin5c (FBtr0345894)"
     5'UTR           6422..6821
                     /label=regulatory region of Dmel U6-3
                     /note="regulatory region of Dmel U6-3"
     misc_feature    6822..6841
                     /label=Dmel white gRNA-5
                     /note="Dmel white gRNA-5"
     misc_RNA        6842..6917
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     3'UTR           6918..7017
                     /label=3'UTR of Dmel U6-3
                     /note="3'UTR of Dmel U6-3"
     polyA_signal    complement(7044..7165)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(7292..7999)
                     /label=mNeonGreen
                     /note="bright monomeric yellow-green fluorescent protein
                     derived from LanYFP (Shaner et al., 2013)"
     regulatory      complement(8000..8270)
                     /label=3xP3 promoter
                     /note="3xP3 promoter"
                     /regulatory_class="promoter"
     misc_feature    8276..9270
                     /label=white homology arm (right)
                     /note="white homology arm (right)"
     promoter        complement(9341..9359)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(9366..9382)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"