Basic Vector Information
- Vector Name:
- MSGV Hu Acceptor
- Antibiotic Resistance:
- Ampicillin
- Length:
- 6699 bp
- Type:
- Retroviral
- Replication origin:
- ori
- Copy Number:
- High Copy
- Promoter:
- MSCV
- Cloning Method:
- Restriction Enzyme
MSGV Hu Acceptor 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 ℃.
MSGV Hu Acceptor vector Sequence
LOCUS MSGV_Hu_Acceptor 6699 bp DNA circular SYN 13-MAY-2021
DEFINITION Retroviral golden gate cloning vector for expression of functional
Human T-Cell Receptor genes.
ACCESSION .
VERSION .
KEYWORDS MSGV Hu Acceptor.
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6699)
AUTHORS Coren LV, Jain S, Trivett MT, Ohlen C, Ott DE
TITLE Production of retroviral constructs for effective transfer and
expression of T-cell receptor genes using Golden Gate cloning.
JOURNAL Biotechniques. 2015 Mar 1;58(3):135-9. doi: 10.2144/000114265.
eCollection 2015.
PUBMED 25757546
REFERENCE 2 (bases 1 to 6699)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 6699)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; doi: "10.2144/000114265";
journalName: "Biotechniques"; date: "2015-03-1- 1"; volume: "58";
issue: "3"; pages: "135-9"
COMMENT SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..6699
/mol_type="other DNA"
/organism="synthetic DNA construct"
LTR 343..518
/label=5' LTR (truncated)
/note="truncated long terminal repeat from Moloney murine
sarcoma virus"
CDS 1021..1428
/label=gag (truncated)
/note="truncated Moloney murine leukemia virus (MMLV) gag
gene lacking the start codon"
misc_feature 1438..1810
/label=pol region
/note="Moloney murine leukemia virus (MMLV) pol region
containing the splice acceptor site"
CDS 1853..2272
/gene="TRAC"
/label=TRAC
/note="T cell receptor alpha chain constant from Homo
sapiens. Accession#: P01848"
CDS 2369..2896
/gene="TRBC1"
/label=TRBC1
/note="T cell receptor beta constant 1 from Homo sapiens.
Accession#: P01850"
misc_feature 2920..2972
/label=MCS
/note="multiple cloning site"
LTR 3016..3530
/label=3' LTR
/note="3' long terminal repeat from murine embryonic stem
cell virus"
protein_bind complement(3702..3718)
/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(3726..3756)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(3771..3792)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(3909..3926)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4080..4668)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(4842..5699)
/label=AmpR
/note="beta-lactamase"
promoter complement(5700..5804)
/label=AmpR promoter
primer_bind 5872..5890
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
primer_bind complement(5928..5950)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind 6050..6069
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 6278..6294
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 6493..6512
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
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