Basic Vector Information
- Vector Name:
- Mm-PylRS-AF-Pyl-tRNACUA
- Antibiotic Resistance:
- Ampicillin
- Length:
- 7401 bp
- Type:
- Protein expression
- Replication origin:
- ori
- Host:
- Mammalian cells
- Selection Marker:
- Hyg
- Promoter:
- U6
- Growth Strain(s):
- DH5a
- Growth Temperature:
- 37℃
Mm-PylRS-AF-Pyl-tRNACUA 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 ℃.
Mm-PylRS-AF-Pyl-tRNACUA vector Sequence
LOCUS 62056_1635 7401 bp DNA circular SYN 01-JAN-1980
DEFINITION synthetic circular DNA.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7401)
AUTHORS .
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..7401
/mol_type="other DNA"
/organism="synthetic DNA construct"
enhancer 47..425
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 426..629
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
promoter 860..878
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
CDS 971..994
/label=FLAG
/note="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
CDS 995..2356
/gene="pylS"
/label=pylS
/note="Pyrrolysine--tRNA ligase from Methanosarcina mazei
(strain ATCC BAA-159 / DSM 3647 / Goe1 / Go1 / JCM 11833 /
OCM 88). Accession#: Q8PWY1"
polyA_signal 2391..2615
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 2661..3089
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 3103..3432
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
CDS 3481..4503
/label=HygR
/note="aminoglycoside phosphotransferase from E. coli"
polyA_signal 4636..4769
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(4806..4822)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(4830..4846)
/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(4854..4884)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(4899..4920)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(5208..5796)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(5970..6827)
/label=AmpR
/note="beta-lactamase"
promoter complement(6828..6932)
/label=AmpR promoter
promoter complement(7030..7270)
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA"
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