AGG1328-pCas13b vector (V016093)

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LOCUS       62056_306        8555 bp DNA     circular SYN 12-AUG-2020
DEFINITION  Cloning vector AGG1328:pCas13b, complete sequence.
ACCESSION   MT119959
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8555)
  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 8555)
  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 8555)
  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..8555
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        140..622
                     /label=hr5 enhancer
                     /note="baculovirus early transcription enhancer"
     misc_feature    1352..4621
                     /gene="PspCas13b-T2A-ZsYellow"
                     /label=PspCas13b
                     /note="PspCas13b"
     misc_feature    4628..4669
                     /gene="PspCas13b-T2A-ZsYellow"
                     /label=super-PK1 NES
                     /note="super-PK1 NES"
     CDS             4679..4705
                     /label=HA
                     /note="HA (human influenza hemagglutinin) epitope tag"
     CDS             4706..4732
                     /label=HA
                     /note="HA (human influenza hemagglutinin) epitope tag"
     CDS             4733..4759
                     /label=HA
                     /note="HA (human influenza hemagglutinin) epitope tag"
     misc_feature    4760..4788
                     /gene="PspCas13b-T2A-ZsYellow"
                     /label=thoseaasigna virus 2A
                     /note="thoseaasigna virus 2A"
     misc_feature    4789..4813
                     /gene="PspCas13b-T2A-ZsYellow"
                     /label=thoseaasigna virus 2A
                     /note="thoseaasigna virus 2A"
     misc_feature    4814..4837
                     /gene="PspCas13b-T2A-ZsYellow"
                     /label=nuclear localization signal
                     /note="nuclear localization signal"
     CDS             4844..5536
                     /label=ZsYellow1
                     /note="Zoanthus yellow fluorescent protein"
     CDS             5546..5566
                     /label=SV40 NLS
                     /note="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
     polyA_signal    complement(5623..5744)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(6151..6739)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6913..7770)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(7771..7875)
                     /label=AmpR promoter
     rep_origin      7902..8357
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     polyA_signal    8488..8536
                     /label=poly(A) signal
                     /note="synthetic polyadenylation signal"
     misc_feature    8550..8555
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from
                     the human alpha-2 globin gene"