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 Vector Map
Plasmid Resuspension 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 ℃.
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"
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