Basic Vector Information
- Vector Name:
- AGG1521-pCHIKVRep1
- Antibiotic Resistance:
- Kanamycin
- Length:
- 5623 bp
- Type:
- Cloning vector
- Replication origin:
- ori
- Source/Author:
- Tng PYL., Carabajal Paladino L, Verkuijl SAN., Purcell J
- Promoter:
- lac
AGG1521-pCHIKVRep1 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 ℃.
AGG1521-pCHIKVRep1 vector Sequence
LOCUS 62056_316 5623 bp DNA circular SYN 12-AUG-2020
DEFINITION Cloning vector AGG1521:pCHIKVRep1, complete sequence.
ACCESSION MT119957
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5623)
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 5623)
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 5623)
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..5623
/mol_type="other DNA"
/organism="synthetic DNA construct"
CDS complement(227..1033)
/codon_start=1
/label=KanR
/note="aminoglycoside phosphotransferase"
/translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
primer_bind 1525..1541
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
CDS 2526..3242
/codon_start=1
/label=EGFP
/note="enhanced GFP"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
CDS 3424..3936
/codon_start=1
/label=Nluc
/note="NanoLuc(R) luciferase"
/translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
RLCERILA"
polyA_signal 4533..4654
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(4679..4695)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(4703..4719)
/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(4727..4757)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(4772..4793)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(join(5081..5623,1..46))
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
This page is informational only.