AGG1201-pZsG vector (V015435)

LOCUS       62056_296        6075 bp DNA     circular SYN 12-AUG-2020
DEFINITION  Cloning vector AGG1201:pZsG, complete sequence.
ACCESSION   MT119955
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6075)
  AUTHORS   Tng PYL., Carabajal Paladino L, Verkuijl SAN., Purcell J, Merits A, 
            Leftwich PT, Fragkoudis R, Noad R, Alphey L.
  TITLE     Cas13b-dependent and Cas13b-independent RNA knockdown of viral 
            sequences in mosquito cells following guide RNA expression
  JOURNAL   Commun Biol 3 (1), 413 (2020)
  PUBMED    32737398
REFERENCE   2  (bases 1 to 6075)
  AUTHORS   Tng PYL., Carabajal Paladino L, Verkuijl S, Purcell J, Merits A, 
            Leftwich PT, Fragkoudis R, Noad R, Alphey L.
  TITLE     Direct Submission
  JOURNAL   Submitted (26-FEB-2020) Arthropod Genetics Group, The Pirbright 
            Institute, Ash Road, Pirbright, Woking, Surrey GU24 0NF, UK
REFERENCE   3  (bases 1 to 6075)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Commun 
            Biol"; date: "2020"; volume: "3"; issue: "1"; pages: "413"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (26-FEB-2020) Arthropod Genetics Group, The Pirbright Institute, Ash
            Road, Pirbright, Woking, Surrey GU24 0NF, UK"
FEATURES             Location/Qualifiers
     source          1..6075
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    41..74
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(414..430)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(438..454)
                     /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(462..492)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(507..528)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(816..1404)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(1578..2435)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(2436..2540)
                     /label=AmpR promoter
     enhancer        2912..3394
                     /label=hr5 enhancer
                     /note="baculovirus early transcription enhancer"
     intron          4138..4207
                     /note="adh"
     misc_feature    4228..4237
                     /label=Kozak sequence
                     /note="Kozak sequence"
     regulatory      4246..4269
                     /gene="ZsGreen"
                     /label=nuclear localization signal
                     /note="nuclear localization signal"
                     /regulatory_class="other"
     CDS             4285..4977
                     /label=ZsGreen1
                     /note="Zoanthus green fluorescent protein"
     CDS             4996..5016
                     /label=SV40 NLS
                     /note="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
     3'UTR           5051..5832
                     /label=K10
                     /note="K10"
     polyA_signal    complement(5841..5962)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"