Rat Smc1a ORF clone (NM_031683) (Cat. No.: V021180)
- Name:
- pAAV-Smc1a(rat)-FLAG
- Accession ID:
- NM_031683.2
- Antibiotic Resistance:
- Ampicillin
- Length:
- 8356 bp
- Type:
- Protein expression
- Replication origin:
- ori
- Host:
- Mammalian cells, Adeno-associated virus
- Promoter:
- CMV
- Fusion Tag:
- FLAG
- Expression Method:
- Transient
- Gene Synonyms:
- SB1.8; SMC-1A; Smc1l1
- Transcript Definition:
- Rattus norvegicus structural maintenance of chromosomes 1A (Smc1a), mRNA
Resources
Plasmid 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 ℃.
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.
Rat Smc1a ORF clone (NM_031683) (Cat. No.: V021180) Sequence
LOCUS pAAV-Smc1a(rat)-FLAG 8356 bp DNA circular SYN 01-JAN-1980
DEFINITION synthetic circular DNA.
ACCESSION V021180
VERSION V021180
KEYWORDS .
SOURCE .
ORGANISM .
.
FEATURES Location/Qualifiers
repeat_region 1..141
/note="inverted terminal repeat of adeno-associated virus
serotype 2"
/rpt_type=inverted
/label="AAV2 ITR"
enhancer 221..524
/note="human cytomegalovirus immediate early enhancer"
/label="CMV enhancer"
promoter 525..727
/note="human cytomegalovirus (CMV) immediate early
promoter"
/label="CMV promoter"
intron 876..1254
/note="fusion between human cytomegalovirus intron A and
human β-globin intron 2"
/label="chimeric intron"
CDS 1343..5041
/label="Smc1a(NM_031683)"
/note="Smc1a(NM_031683)"
/gene="Smc1a"
CDS 5042..5065
/codon_start=1
/product="FLAG® epitope tag, followed by an enterokinase
cleavage site"
/transl_table=1
/translation="DYKDDDDK"
/label="FLAG"
polyA_signal 5103..5579
/note="human growth hormone polyadenylation signal"
/label="hGH poly(A) signal"
repeat_region 5619..5759
/note="inverted terminal repeat of adeno-associated virus
serotype 2"
/rpt_type=inverted
/label="AAV2 ITR"
rep_origin 5834..6289
/direction=RIGHT
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
/label="f1 ori"
promoter 6571..6675
/gene="bla"
/label="AmpR promoter"
CDS 6676..7536
/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"
rep_origin 7707..8295
/direction=RIGHT
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
/label="ori"