Basic Vector Information
- Vector Name:
- pLeo410
- Antibiotic Resistance:
- Kanamycin
- Length:
- 6580 bp
- Type:
- Expression vector
- Replication origin:
- ori
- Source/Author:
- Scheller L, Strittmatter T, Fuchs D, Bojar D, Fussenegger M.
pLeo410 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 ℃.
pLeo410 vector Sequence
LOCUS 40924_28007 6580 bp DNA circular SYN 18-DEC-2018
DEFINITION Expression vector pLeo410, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6580)
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 6580)
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 6580)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 6580)
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..6580
/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)"
misc_feature 846..902
/label=GCN4 tag
/note="GCN4 tag"
CDS 846..902
/codon_start=1
/product="GCN4 peptide optimized to improve solubility
while preserving binding to an scFv-GFP fusion protein
(Tanenbaum et al., 2014)"
/label=GCN4_v4
/translation="EELLSKNYHLENEVARLKK"
misc_feature 918..974
/label=GCN4 tag
/note="GCN4 tag"
CDS 918..974
/codon_start=1
/product="GCN4 peptide optimized to improve solubility
while preserving binding to an scFv-GFP fusion protein
(Tanenbaum et al., 2014)"
/label=GCN4_v4
/translation="EELLSKNYHLENEVARLKK"
misc_feature 963..965
/label=R
/note="R"
misc_feature 990..1046
/label=GCN4 tag
/note="GCN4 tag"
CDS 990..1046
/codon_start=1
/product="GCN4 peptide optimized to improve solubility
while preserving binding to an scFv-GFP fusion protein
(Tanenbaum et al., 2014)"
/label=GCN4_v4
/translation="EELLSKNYHLENEVARLKK"
misc_feature 1062..1118
/label=GCN4 tag
/note="GCN4 tag"
CDS 1062..1118
/codon_start=1
/product="GCN4 peptide optimized to improve solubility
while preserving binding to an scFv-GFP fusion protein
(Tanenbaum et al., 2014)"
/label=GCN4_v4
/translation="EELLSKNYHLENEVARLKK"
misc_feature 1134..1190
/label=GCN4 tag
/note="GCN4 tag"
CDS 1134..1190
/codon_start=1
/product="GCN4 peptide optimized to improve solubility
while preserving binding to an scFv-GFP fusion protein
(Tanenbaum et al., 2014)"
/label=GCN4_v4
/translation="EELLSKNYHLENEVARLKK"
misc_feature 1206..1262
/label=GCN4 tag
/note="GCN4 tag"
CDS 1206..1262
/codon_start=1
/product="GCN4 peptide optimized to improve solubility
while preserving binding to an scFv-GFP fusion protein
(Tanenbaum et al., 2014)"
/label=GCN4_v4
/translation="EELLSKNYHLENEVARLKK"
misc_feature 1278..1334
/label=GCN4 tag
/note="GCN4 tag"
CDS 1278..1334
/codon_start=1
/product="GCN4 peptide optimized to improve solubility
while preserving binding to an scFv-GFP fusion protein
(Tanenbaum et al., 2014)"
/label=GCN4_v4
/translation="EELLSKNYHLENEVARLKK"
misc_feature 1350..1406
/label=GCN4 tag
/note="GCN4 tag"
CDS 1350..1406
/codon_start=1
/product="GCN4 peptide optimized to improve solubility
while preserving binding to an scFv-GFP fusion protein
(Tanenbaum et al., 2014)"
/label=GCN4_v4
/translation="EELLSKNYHLENEVARLKK"
CDS 1425..1445
/label=TEV site
/note="tobacco etch virus (TEV) protease recognition and
cleavage site"
CDS 1461..1478
/label=6xHis
/note="6xHis affinity tag"
terminator 1545..1592
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
rep_origin 1629..2084
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
CDS complement(2180..2992)
/label=KanR
/note="aminoglycoside phosphotransferase"
rep_origin 3114..3702
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
misc_feature complement(3888..4030)
/label=bom
/note="basis of mobility region from pBR322"
CDS complement(4135..4323)
/label=rop
/note="Rop protein, which maintains plasmids at low copy
number"
protein_bind complement(5098..5119)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
CDS complement(5135..6214)
/label=lacI
/note="lac repressor"
promoter complement(6215..6292)
/label=lacI promoter
This page is informational only.