Basic Vector Information
- Vector Name:
- GLP1R-Tango
- Antibiotic Resistance:
- Ampicillin
- Length:
- 8027 bp
- Type:
- Mammalian Expression
- Replication origin:
- ori
- Selection Marker:
- Neomycin (select with G418)
- Copy Number:
- High Copy
- Promoter:
- CMV
- Cloning Method:
- Restriction Enzyme
- 5' Primer:
- CMV-F
- 3' Primer:
- Tet-R
GLP1R-Tango 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 ℃.
GLP1R-Tango vector Sequence
LOCUS GLP1R-Tango. 8027 bp DNA circular SYN 13-MAY-2021
DEFINITION Expression of G protein-coupled receptors for PRESTO-Tango: parallel
receptorome expression and screening via transcriptional output,
with transcriptional activation following arrestin translocation.
ACCESSION .
VERSION .
KEYWORDS GLP1R-Tango.
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8027)
AUTHORS Kroeze WK, Sassano MF, Huang XP, Lansu K, McCorvy JD, Giguere PM,
Sciaky N, Roth BL
TITLE PRESTO-Tango as an open-source resource for interrogation of the
druggable human GPCRome.
JOURNAL Nat Struct Mol Biol. 2015 May;22(5):362-9. doi: 10.1038/nsmb.3014.
Epub 2015 Apr 20.
PUBMED 25895059
REFERENCE 2 (bases 1 to 8027)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 8027)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; doi: "10.1038/nsmb.3014";
journalName: "Nat Struct Mol Biol"; date: "2015-05"; volume: "22";
issue: "5"; pages: "362-9"
COMMENT SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..8027
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind complement(44..63)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 235..614
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 615..818
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
promoter 863..881
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 1073..2461
/gene="GLP1R"
/label=GLP1R
/note="Glucagon-like peptide 1 receptor from Homo sapiens.
Accession#: P43220"
CDS 2582..3580
/label=tTA
/note="tetracycline-controlled transactivator, comprising a
fusion of the tetracycline repressor TetR with the
C-terminal activation domain of herpes simplex virus VP16"
polyA_signal 3627..3851
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 3897..4325
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 4339..4668
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
CDS 4735..5526
/label=NeoR/KanR
/note="aminoglycoside phosphotransferase"
polyA_signal 5703..5836
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(5873..5889)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(5897..5913)
/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(5921..5951)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(5966..5987)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(6104..6121)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(6275..6860)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(7034..7891)
/label=AmpR
/note="beta-lactamase"
promoter complement(7892..7996)
/label=AmpR promoter
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