Basic Vector Information
- Vector Name:
- AGG1201-pZsG
- Antibiotic Resistance:
- Ampicillin
- Length:
- 6075 bp
- Type:
- Cloning vector
- Replication origin:
- ori
- Source/Author:
- Tng PYL., Carabajal Paladino L, Verkuijl SAN., Purcell J
- Promoter:
- IE1
AGG1201-pZsG 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 ℃.
AGG1201-pZsG vector Sequence
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"
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