Basic Vector Information
- Vector Name:
- no-repeats
- Antibiotic Resistance:
- Ampicillin
- Length:
- 12218 bp
- Type:
- Cloning vector
- Replication origin:
- ori
- Source/Author:
- Akmammedov A, Geigges M, Paro R.
- Promoter:
- hsp70
no-repeats 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 ℃.
no-repeats vector Sequence
LOCUS 40924_2249 12218 bp DNA circular SYN 18-DEC-2018
DEFINITION Cloning vector no-repeats, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 12218)
AUTHORS Akmammedov A, Geigges M, Paro R.
TITLE Single vector non-leaky gene expression system for Drosophila
melanogaster
JOURNAL Sci Rep 7 (1), 6899 (2017)
PUBMED 28761084
REFERENCE 2 (bases 1 to 12218)
AUTHORS Akmammedov A, Geigges M, Paro R.
TITLE Direct Submission
JOURNAL Submitted (04-MAY-2017) Biosystem Science and Engineering, ETH
Zurich, Mattenstrasse 26, Basel 4058, Switzerland
REFERENCE 3 (bases 1 to 12218)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 12218)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Sci Rep";
date: "2017"; volume: "7"; issue: "1"; pages: "6899"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(04-MAY-2017) Biosystem Science and Engineering, ETH Zurich,
Mattenstrasse 26, Basel 4058, Switzerland"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..12218
/mol_type="other DNA"
/organism="synthetic DNA construct"
rep_origin complement(3..458)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind 600..616
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
promoter 623..641
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
gene 666..4802
/label=mini-white
/note="This modified version of the white gene lacks part
of the first intron."
protein_bind 4842..5227
/label=gypsy insulator
/note="chromatin insulator from Drosophila"
polyA_signal complement(5235..5661)
/label=hsp70 poly(A)
/note="polyadenlyation signal from the Drosophila
melanogaster hsp70Ab gene"
CDS complement(5680..8724)
/label=lacZ
/note="beta-galactosidase"
promoter complement(8846..9294)
/label=hsp70 promoter
/note="Drosophila melanogaster hsp70Bb promoter"
regulatory 9316..9702
/label=gypsy
/note="gypsy"
/regulatory_class="insulator"
protein_bind 9317..9702
/label=gypsy insulator
/bound_moiety="Su(Hw), CP190, and Mod(mdg4)67.2"
/note="chromatin insulator from Drosophila"
protein_bind 9742..9811
/label=attB
/note="attB site for the phi-C31 integrase (Groth et al.,
2000)"
promoter complement(10030..10048)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(10069..10085)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 10093..10109
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/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(10117..10147)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(10162..10183)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(10471..11059)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(11233..12090)
/label=AmpR
/note="beta-lactamase"
promoter complement(12091..12195)
/label=AmpR promoter
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