Human EHMT2 ORF clone (NM_001289413) (Cat. No.: V046024)
- Name:
- pBiT2.1-EHMT2(human)
- Accession ID:
- NM_001289413.2
- Antibiotic Resistance:
- Ampicillin
- Length:
- 7102 bp
- Type:
- Protein expression
- Replication origin:
- ori
- Host:
- Mammalian cells
- Copy Number:
- High
- Promoter:
- TK
- 5' Primer:
- HSV TK-F1
- Growth Strain(s):
- DH5a
- Growth Temperature:
- 37℃
- Expression Method:
- Transient
- Gene Synonyms:
- BAT8; C6orf30; G9A; GAT8; KMT1C; NG36
- Transcript Definition:
- Homo sapiens euchromatic histone lysine methyltransferase 2 (EHMT2), transcript variant 1, 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 EHMT2 ORF clone (NM_001289413) (Cat. No.: V046024) Sequence
LOCUS pBiT2.1-EHMT2(human) 7102 bp DNA circular SYN 01-JAN-1980
DEFINITION synthetic circular DNA.
ACCESSION V046024
VERSION V046024
KEYWORDS .
SOURCE .
ORGANISM .
.
FEATURES Location/Qualifiers
promoter 28..779
/note="herpes simplex virus thymidine kinase promoter"
/label="HSV TK promoter"
regulatory 809..818
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
/label="Kozak sequence"
CDS 890..4588
/label="EHMT2(NM_001289413)"
/note="EHMT2(NM_001289413)"
/gene="EHMT2"
polyA_signal 4656..4777
/note="SV40 polyadenylation signal"
/label="SV40 poly(A) signal"
rep_origin complement(5196..5784)
/direction=LEFT
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
/label="ori"
CDS complement(5984..6844)
/codon_start=1
/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"
polyA_signal 6949..6997
/note="synthetic polyadenylation signal"
/label="poly(A) signal"
misc_feature 7011..7102
/note="RNA polymerase II transcriptional pause signal from
the human α2 globin gene"
/label="pause site"