Basic Vector Information
- Vector Name:
- AGG1328-pCas13b
- Antibiotic Resistance:
- Ampicillin
- Length:
- 8555 bp
- Type:
- Cloning vector
- Replication origin:
- ori
- Source/Author:
- Tng PYL., Carabajal Paladino L, Verkuijl SAN., Purcell J
- Promoter:
- IE1
AGG1328-pCas13b vector Vector Map
Plasmid Resuspension Protocol:
1. Centrifuge at 5,000×g for 5 min.
2. Carefully open the tube and add 20 μl of sterile water to dissolve the DNA.
3. Close the tube and incubate for 10 minutes at room temperature.
4. Briefly vortex the tube and then do a quick spin to concentrate the liquid at the bottom. Speed is less than 5000×g.
5.Store the plasmid at -20 ℃.
AGG1328-pCas13b vector Sequence
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"
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