pVG363_Aste-U6a-Swap4-gRNA vector (V016117)

LOCUS       62056_22575        4862 bp DNA     circular SYN 07-OCT-2020
DEFINITION  Cloning vector pVG363_Aste-U6a-Swap4-gRNA, complete sequence.
ACCESSION   MW030450
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4862)
  AUTHORS   Adolfi A, Gantz V, Jasinskiene N, Lee H-F., Hwang K, Terradas G, 
            Bulger E, Ramaiah A, Bennett J, Emerson JJ, Marshall J, Bier E.
  TITLE     Efficient population modification gene-drive rescue system in the 
            malaria mosquito Anopheles stephensi
  JOURNAL   Nat Commun (2020) In press
REFERENCE   2  (bases 1 to 4862)
  AUTHORS   Adolfi A, Gantz V, Jasinskiene N, Lee H-F., Hwang K, Terradas G, 
            Bulger E, Ramaiah A, Bennett J, Emerson JJ, Marshall J, Bier E.
  TITLE     Direct Submission
  JOURNAL   Submitted (22-SEP-2020) Developmental and Stem Cell Biology, 
            University of California, San Francisco, 1650 Owens St., San 
            Francisco, CA 94158, USA
REFERENCE   3  (bases 1 to 4862)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat Commun 
            (2020) In press"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (22-SEP-2020) Developmental and Stem Cell Biology, University of 
            California, San Francisco, 1650 Owens St., San Francisco, CA 94158, 
            USA"
FEATURES             Location/Qualifiers
     source          1..4862
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(333..1124)
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     rep_origin      complement(1468..1896)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     2037..2053
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2060..2078
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_RNA        2597..2672
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(3153..3169)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(3177..3193)
                     /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)."
     promoter        complement(3201..3231)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(3246..3267)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(3555..4143)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(4317..4862,1..312))
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"