Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V000064 | pCMV(CAT)T7-SB100 | In stock, 1 week for quality controls |
Buy one, get one free! |
Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.
Basic Vector Information
- Vector Name:
- pCMV(CAT)T7-SB100
- Antibiotic Resistance:
- Chloramphenicol
- Length:
- 4755 bp
- Type:
- Mammalian Expression
- Replication origin:
- ori
- Copy Number:
- High Copy
- Promoter:
- CMV
- Cloning Method:
- Restriction Enzyme
- 5' Primer:
- CACCAAAATCAACGGGACTT
pCMV(CAT)T7-SB100 vector Map
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.
pCMV(CAT)T7-SB100 vector Sequence
LOCUS 40924_11601 4755 bp DNA circular SYN 09-SEP-2021
DEFINITION synthetic circular DNA.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4755)
AUTHORS Mates L, Chuah MK, Belay E, Jerchow B, Manoj N, Acosta-Sanchez A,
Grzela DP, Schmitt A, Becker K, Matrai J, Ma L, Samara-Kuko E,
Gysemans C, Pryputniewicz D, Miskey C, Fletcher B, VandenDriessche
T, Ivics Z, Izsvak Z
TITLE Molecular evolution of a novel hyperactive Sleeping Beauty
transposase enables robust stable gene transfer in vertebrates.
JOURNAL Nat Genet. 2009 Jun;41(6):753-61. Epub 2009 May 3.
PUBMED 19412179
REFERENCE 2 (bases 1 to 4755)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 4755)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nat
Genet."; date: "2009-06"; volume: "41(6)"; pages: "753-61. Epub 2009
May 3"
COMMENT SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..4755
/mol_type="other DNA"
/organism="synthetic DNA construct"
enhancer 30..333
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 334..537
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 534..558
/label=LNCX
/note="Human CMV promoter, forward primer"
intron 675..771
/label=SV40 intron
/note="modified SV40 intron with splice donor and acceptor
sites"
promoter 828..846
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 857..1876
/codon_start=1
/label=SB100X
/note="hyperactive variant of the Sleeping Beauty
transposase (Mates et al., 2009)"
/translation="MGKSKEISQDLRKRIVDLHKSGSSLGAISKRLAVPRSSVQTIVRK
YKHHGTTQPSYRSGRRRVLSPRDERTLVRKVQINPRTTAKDLVKMLEETGTKVSISTVK
RVLYRHNLKGHSARKKPLLQNRHKKARLRFATAHGDKDRTFWRNVLWSDETKIELFGHN
DHRYVWRKKGEACKPKNTIPTVKHGGGSIMLWGCFAAGGTGALHKIDGIMDAVQYVDIL
KQHLKTSVRKLKLGRKWVFQHDNDPKHTSKVVAKWLKDNKVKVLEWPSQSPDLNPIENL
WAELKKRVRARRPTNLTQLHQLCQEEWAKIHPNYCGKLVEGYPKRLTQVKQFKGNATKY
"
polyA_signal complement(1896..2030)
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(2141..2157)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(2165..2181)
/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(2189..2219)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(2234..2255)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(2372..2389)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(2543..3131)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(3863..4519)
/codon_start=1
/label=CmR
/note="chloramphenicol acetyltransferase"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
promoter complement(4520..4622)
/label=cat promoter
/note="promoter of the E. coli cat gene encoding
chloramphenicol acetyltransferase"