Basic Vector Information
murine TCR OTI-2A.pMIG II 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 ℃.
murine TCR OTI-2A.pMIG II vector Sequence
LOCUS murine_TCR_OTI-2 8327 bp DNA circular SYN 13-MAY-2021
DEFINITION retroviral expression of both TCRalpha and TCRbeta using a single
vector.
ACCESSION .
VERSION .
KEYWORDS murine TCR OTI-2A.pMIG II.
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8327)
AUTHORS Holst J, Szymczak-Workman AL, Vignali KM, Burton AR, Workman CJ,
Vignali DA
TITLE Generation of T-cell receptor retrogenic mice.
JOURNAL Nat Protoc. 2006;1(1):406-17.
PUBMED 17406263
REFERENCE 2 (bases 1 to 8327)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 8327)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nat
Protoc."; date: "2006"; volume: "1(1)"; pages: "406-17"
COMMENT SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..8327
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 1..515
/label=MSCV
/note="Murine embryonic stem cell virus promoter including
the enhancer and promoter region of PCMV virus and 5'
untranslated region of dl-587rev retrovirus"
misc_feature 579..920
/label=MESV Psi
/note="packaging signal of murine embryonic stem cell
virus"
CDS 987..1403
/label=gag (truncated)
/note="truncated Moloney murine leukemia virus (MMLV) gag
gene lacking the start codon"
CDS 1837..2244
/gene="Trac"
/label=Trac
/note="T-cell receptor alpha chain constant from Mus
musculus. Accession#: P01849"
CDS 2248..2301
/label=T2A
/note="2A peptide from Thosea asigna virus capsid protein"
misc_feature 3275..3836
/label=IRES
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
CDS 3839..4555
/label=EGFP
/note="enhanced GFP"
LTR 4720..5234
/label=3' LTR
/note="3' long terminal repeat from murine embryonic stem
cell virus"
protein_bind complement(5396..5412)
/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(5420..5450)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(5465..5486)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(5603..5620)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(5774..6362)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(6536..7393)
/label=AmpR
/note="beta-lactamase"
promoter complement(7394..7498)
/label=AmpR promoter
primer_bind 7566..7584
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
primer_bind complement(7622..7644)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind 7744..7763
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 7957..7979
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 8107..8126
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
LTR 8326..8327
/label=5' LTR
/note="5' long terminal repeat from murine embryonic stem
cell virus"
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