Human SRP9 ORF clone (NM_003133) (Cat. No.: V018243)
Note: 1 NT, 1 AA Mismatches
- Name:
- pCMV-SPORT6-SRP9(human)-FLAG-VN173
- Accession ID:
- NM_003133.6
- Antibiotic Resistance:
- Ampicillin
- Length:
- 5879 bp
- Type:
- Protein expression
- Replication origin:
- ori
- Host:
- Mammalian cells
- Promoter:
- CMV
- Fusion Tag:
- FLAG-VN173
- Expression Method:
- Transient
- Gene Synonyms:
- ALURBP
- Transcript Definition:
- Homo sapiens signal recognition particle 9 (SRP9), transcript variant 2, mRNA
Resources
Plasmid Protocol
1. Centrifuge at 5,000×g for 5 min.
2. Carefully open the tube and add sterile water to dissolve the DNA: add 20 μl for 5 μg plasmid, and 100 μl for 100 μg plasmid.
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 ℃.
6. The concentration of plasmid re-measurement sometimes differs from the nominal value, which may be due to the position of the lyophilized plasmid in the tube, the efficiency of the re-dissolution, the measurement bias, and adsorption on the wall of the tube, therefore, it is recommended to transform and extract the plasmid before using it
General Plasmid Transform Protocol
1. Take one 100μl of the competent cells and thaw it on ice for 10min, add 2μl of plasmid, then ice bath for 30min, then heat-shock it at 42℃ for 60s, do not stir, and then ice bath for 2min.
2. Add 900μl of LB liquid medium without antibiotics, and incubate at 37℃ for 45min (30℃ for 1-1.5 hours) with 180rpm shaking.
3. Centrifuge at 6000rpm for 5min, leave only 100μl of supernatant to resuspend the bacterial precipitate and spread it onto the target plasmid-resistant LB plate.
4. Invert the plate and incubate at 37℃ for 14h, or at 30℃ for 20h.
5. Pick a single colony into LB liquid medium, add the corresponding antibiotics, incubate at 220rpm for 14h, and extract the plasmid according to the experimental needs and the instructions of the plasmid extraction kit.
Human SRP9 ORF clone (NM_003133) (Cat. No.: V018243) Sequence
LOCUS V018243 5879 bp DNA circular SYN 01-JAN-1980
DEFINITION synthetic circular DNA.
ACCESSION V018243
VERSION V018243
KEYWORDS .
SOURCE .
ORGANISM .
.
FEATURES Location/Qualifiers
primer_bind 141..157
/note="common sequencing primer, one of multiple similar
variants"
/label="M13 fwd"
enhancer 318..697
/note="human cytomegalovirus immediate early enhancer"
/label="CMV enhancer"
promoter 698..901
/note="human cytomegalovirus (CMV) immediate early
promoter"
/label="CMV promoter"
CDS 935..958
/codon_start=1
/product="FLAG® epitope tag, followed by an enterokinase
cleavage site"
/transl_table=1
/translation="DYKDDDDK"
/label="FLAG"
RBS 1230..1238
/note="full consensus sequence for ribosome-binding sites
upstream of start codons in E. coli; complementary to a
region in the 3' end of the 16S rRNA (Chen et al., 1994)"
/label="Shine-Dalgarno sequence"
CDS 1397..1654
/label="SRP9(NM_003133)"
/note="SRP9(NM_003133)"
/gene="SRP9"
CDS 1691..2209
/codon_start=1
/product="N-terminal fragment of mVenus for use in
bimolecular fluorescence complementation (BiFC) (Kodama and
Hu, 2010)"
/transl_table=1
/translation="M,V,SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKL
TLKLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFK
DDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNG
IKANFKIRHNIE"
/label="VN173"
polyA_signal 2220..2842
/note="human growth hormone polyadenylation signal"
/label="hGH poly(A) signal"
promoter 2871..3200
/note="SV40 enhancer and early promoter"
/label="SV40 promoter"
promoter complement(3239..3257)
/note="promoter for bacteriophage T7 RNA polymerase"
/label="T7 promoter"
primer_bind complement(3271..3287)
/note="common sequencing primer, one of multiple similar
variants"
/label="M13 rev"
protein_bind 3295..3311
/bound_moiety="lac repressor encoded by lacI"
/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-β-D-thiogalactopyranoside (IPTG)."
/label="lac operator"
promoter complement(3319..3349)
/note="promoter for the E. coli lac operon"
/label="lac promoter"
protein_bind 3364..3385
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
/label="CAP binding site"
rep_origin complement(3673..4261)
/direction=LEFT
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
/label="ori"
CDS complement(4432..5292)
/codon_start=1
/gene="bla"
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/product="β-lactamase"
/transl_table=1
/translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY
IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY
SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR
WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS
ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA
SLIKHW*"
/label="AmpR"
promoter complement(5293..5397)
/gene="bla"
/label="AmpR promoter"
rep_origin 5424..5852
/direction=RIGHT
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
/label="f1 ori"