Basic Vector Information
- Vector Name:
- pleo127
- Antibiotic Resistance:
- Kanamycin
- Length:
- 6670 bp
- Type:
- Expression vector
- Replication origin:
- ori
- Source/Author:
- Scheller L, Strittmatter T, Fuchs D, Bojar D, Fussenegger M.
pleo127 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 ℃.
pleo127 vector Sequence
LOCUS 40924_28002 6670 bp DNA circular SYN 18-DEC-2018
DEFINITION Expression vector pleo127, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6670)
AUTHORS Scheller L, Strittmatter T, Fuchs D, Bojar D, Fussenegger M.
TITLE Generalized extracellular molecule sensor platform for programming
cellular behavior
JOURNAL Nat. Chem. Biol. 14 (7), 723-729 (2018)
PUBMED 29686358
REFERENCE 2 (bases 1 to 6670)
AUTHORS Scheller L, Strittmatter T.
TITLE Direct Submission
JOURNAL Submitted (06-NOV-2017) D-BSSE, ETH Zurich, Mattenstrasse 26, Basel
4058, Switzerland
REFERENCE 3 (bases 1 to 6670)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 6670)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nat. Chem.
Biol."; date: "2018"; volume: "14"; issue: "7"; pages: "723-729"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(06-NOV-2017) D-BSSE, ETH Zurich, Mattenstrasse 26, Basel 4058,
Switzerland"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..6670
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 21..39
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 40..64
/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)."
RBS 79..101
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
misc_feature 84..100
/label=T7_transl_en_RBS
/note="T7_transl_en_RBS"
CDS 108..815
/label=mCherry
/note="monomeric derivative of DsRed fluorescent protein
(Shaner et al., 2004)"
CDS 822..1529
/codon_start=1
/product="monomeric derivative of DsRed fluorescent protein
(Shaner et al., 2004)"
/label=mCherry
/note="mammalian codon-optimized"
/translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
GRHSTGGMDELYK"
misc_feature 822..1523
/label=mCherry
/note="mCherry"
CDS 1551..1568
/label=6xHis
/note="6xHis affinity tag"
terminator 1635..1682
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
rep_origin 1719..2174
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
CDS complement(2270..3082)
/label=KanR
/note="aminoglycoside phosphotransferase"
rep_origin 3204..3792
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
misc_feature complement(3978..4120)
/label=bom
/note="basis of mobility region from pBR322"
CDS complement(4225..4413)
/label=rop
/note="Rop protein, which maintains plasmids at low copy
number"
protein_bind complement(5188..5209)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
CDS complement(5225..6304)
/label=lacI
/note="lac repressor"
promoter complement(6305..6382)
/label=lacI promoter
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