Basic Vector Information
- Vector Name:
- pEN-2xChimera
- Antibiotic Resistance:
- Ampicillin
- Length:
- 4236 bp
- Type:
- Cloning vector
- Replication origin:
- ori
- Source/Author:
- Durr J, Papareddy R, Nakajima K, Gutierrez-Marcos J.
- Promoter:
- SP6
pEN-2xChimera 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 ℃.
pEN-2xChimera vector Sequence
LOCUS 40924_17399 4236 bp DNA circular SYN 17-DEC-2018
DEFINITION Cloning vector pEN-2xChimera, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4236)
AUTHORS Durr J, Papareddy R, Nakajima K, Gutierrez-Marcos J.
TITLE Highly efficient heritable targeted deletions of gene clusters and
non-coding regulatory regions in Arabidopsis using CRISPR/Cas9
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 4236)
AUTHORS Durr J, Gutierrez-Marcos J.
TITLE Direct Submission
JOURNAL Submitted (19-JAN-2017) School of Life Sciences, University of
Warwick, Gibbet Hill Campus, Coventry, West Midlands CV4 7AL, United
Kingdom
REFERENCE 3 (bases 1 to 4236)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 4236)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName:
"Unpublished"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(19-JAN-2017) School of Life Sciences, University of Warwick, Gibbet
Hill Campus, Coventry, West Midlands CV4 7AL, United Kingdom"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..4236
/mol_type="other DNA"
/organism="synthetic DNA construct"
regulatory 33..42
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
regulatory 37..46
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
protein_bind 52..151
/label=attL1
/note="recombination site for the Gateway(R) LR reaction"
regulatory 152..539
/label=U6-26
/note="U6-26"
/regulatory_class="promoter"
misc_RNA 558..633
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
protein_bind complement(645..669)
/gene="mutant version of attB"
/label=attB2
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
regulatory 681..1068
/label=U6-26
/note="U6-26"
/regulatory_class="promoter"
misc_feature 1085..1201
/label=sgRNA scaffold
/note="sgRNA scaffold"
misc_RNA 1085..1160
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
protein_bind complement(1172..1196)
/gene="mutant version of attB"
/label=attB2
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
protein_bind complement(1226..1325)
/label=attL2
/note="recombination site for the Gateway(R) LR reaction"
promoter complement(1361..1379)
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind complement(1397..1413)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(1421..1437)
/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(1445..1475)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(1490..1511)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(1799..2387)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(2561..3418)
/label=AmpR
/note="beta-lactamase"
promoter complement(3419..3523)
/label=AmpR promoter
rep_origin complement(3601..4056)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind 4197..4213
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
promoter 4220..4236
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
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