Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V013396 | Ai9 | 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:
- Ai9
- Antibiotic Resistance:
- Ampicillin
- Length:
- 17251 bp
- Type:
- Protein expression
- Replication origin:
- ori
- Host:
- Mammalian cells
- Selection Marker:
- Neo/G418
- Promoter:
- mPGK
- Growth Strain(s):
- Stbl3
- Growth Temperature:
- 37℃
Ai9 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.
Ai9 vector Sequence
LOCUS 62056_361 17251 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 17251)
AUTHORS .
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..17251
/mol_type="other DNA"
/organism="synthetic DNA construct"
intron 153..1169
/label=chimeric intron
/note="chimera between introns from chicken beta-actin and
rabbit beta-globin"
protein_bind 1245..1278
/label=FRT (minimal)
/note="supports FLP-mediated excision but not integration
(Turan and Bode, 2011)"
protein_bind 1302..1335
/label=loxP
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (ATGTATGC) (Shaw et al., 2021)."
polyA_signal 1502..1636
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
CDS 2033..3460
/codon_start=1
/label=tdTomato
/note="tandem dimeric (pseudo-monomeric) derivative of
DsRed (Shaner et al., 2004)"
/translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
HLFLGHGTGSTGSGSSGTASSEDNNMAVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPY
EGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVM
NFEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGV
LKGEIHQALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYE
RSEGRHHLFLYGMDELYK"
misc_feature 3529..4117
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(4120..4136)
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
polyA_signal 4144..4351
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
protein_bind 4359..4392
/label=attB
/note="minimal attB site for the phi-C31 integrase (Groth
et
al., 2000)"
promoter 4414..4913
/label=PGK promoter
/note="mouse phosphoglycerate kinase 1 promoter"
CDS 4973..5773
/codon_start=1
/label=NeoR/KanR
/note="aminoglycoside phosphotransferase from Tn5"
/translation="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ
GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD
LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
EHQGLAPAELFARLKARMPDGDDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ
DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
polyA_signal 5786..6247
/label=PGK poly(A) signal
/note="mouse phosphoglycerate kinase 1 polyadenylation
signal"
misc_feature 6318..7128
/label=Rosa26 right arm
/note="right homology arm from intron 1 of the mouse Rosa26
gene (Soriano, 1999)"
promoter 10703..11202
/label=PGK promoter
/note="mouse phosphoglycerate kinase 1 promoter"
CDS 11229..11801
/codon_start=1
/label=DTA
/note="diphtheria toxin fragment A"
/translation="DDVVDSSKSFVMENFSSYHGTKPGYVDSIQKGIQKPKSGTQGNYD
DDWKGFYSTDNKYDAAGYSVDNENPLSGKAGGVVKVTYPGLTKVLALKVDNAETIKKEL
GLSLTEPLMEQVGTEEFIKRFGDGASRVVLSLPFAEGSSSVEYINNWEQAKALSVELEI
NFETRGKRGQDAMYEYMAQACAGNRVRR"
intron 11878..11943
/label=small t intron
/note="SV40 (simian virus 40) small t antigen intron"
polyA_signal 12017..12241
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
promoter complement(12303..12321)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
promoter complement(12390..12420)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(12435..12456)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(12744..13332)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(13506..14363)
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(14364..14468)
/label=AmpR promoter
rep_origin 14495..14950
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind 15091..15107
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
promoter 15117..15135
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
misc_feature 15932..16725
/label=Rosa26 left arm
/note="left homology arm from intron 1 of the mouse Rosa26
gene (Soriano, 1999)"
enhancer 16821..17124
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"