pTE4254
Catalog No. PVT10992
Packing 2ug
pTE4254 Information
Function Mammal Editing plasmids
Resistance Amp
Screen Neo/G418
Strain Stbl3
Culture temperature 37degrees centigrade
Replicon pUC
Copy high
Promoter U6
pTE4254 Description
PTE4254 is a plasmid expressing CAS9 and gRNA in mammalian cells.
pTE4254 Multiple cloning site
pTE4254 Sequence
LOCUS Exported 9204 bp ds-DNA circular SYN 20-DEC-2017
DEFINITION synthetic circular DNA
FEATURES Location/Qualifiers
source 1..9204
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 18..397
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 398..601
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
CDS 716..4078
/codon_start=1
/product="Cas9 (Csn1) endonuclease from the Streptococcus
thermophilus Type II CRISPR/Cas system"
/label=St1Cas9
/note="generates RNA-guided double strand breaks in DNA"
/translation="MSDLVLGLDIGIGSVGVGILNKVTGEIIHKNSRIFPAAQAENNLV
RRTNRQGRRLARRKKHRRVRLNRLFEESGLITDFTKISINLNPYQLRVKGLTDELSNEE
LFIALKNMVKHRGISYLDDASDDGNSSVGDYAQIVKENSKQLETKTPGQIQLERYQTYG
QLRGDFTVEKDGKKHRLINVFPTSAYRSEALRILQTQQEFNPQITDEFINRYLEILTGK
RKYYHGPGNEKSRTDYGRYRTSGETLDNIFGILIGKCTFYPDEFRAAKASYTAQEFNLL
NDLNNLTVPTETKKLSKEQKNQIINYVKNEKAMGPAKLFKYIAKLLSCDVADIKGYRID
KSGKAEIHTFEAYRKMKTLETLDIEQMDRETLDKLAYVLTLNTEREGIQEALEHEFADG
SFSQKQVDELVQFRKANSSIFGKGWHNFSVKLMMELIPELYETSEEQMTILTRLGKQKT
TSSSNKTKYIDEKLLTEEIYNPVVAKSVRQAIKIVNAAIKEYGDFDNIVIEMARETNED
DEKKAIQKIQKANKDEKDAAMLKAANQYNGKAELPHSVFHGHKQLATKIRLWHQQGERC
LYTGKTISIHDLINNSNQFEVDHILPLSITFDDSLANKVLVYATANQEKGQRTPYQALD
SMDDAWSFRELKAFVRESKTLSNKKKEYLLTEEDISKFDVRKKFIERNLVDTLYASRVV
LNALQEHFRAHKIDTKVSVVRGQFTSQLRRHWGIEKTRDTYHHHAVDALIIAASSQLNL
WKKQKNTLVSYSEDQLLDIETGELISDDEYKESVFKAPYQHFVDTLKSKEFEDSILFSY
QVDSKFNRKISDATIYATRQAKVGKDKADETYVLGKIKDIYTQDGYDAFMKIYKKDKSK
FLMYRHDPQTFEKVIEPILENYPNKQINDKGKEVPCNPFLKYKEEHGYIRKYSKKGNGP
EIKSLKYYDSKLGNHIDITPKDSNNKVVLQSVSPWRADVYFNKTTGKYEILGLKYADLQ
FDKGTGTYKISQEKYNDIKKKEGVDSDSEFKFTLYKNDLLLVKDTETKEQQLFRFLSRT
MPKQKHYVELKPYDKQKFEGGEALIKVLGNVANSGQCKKGLGKSNISIYKVRTDVLGNQ
HIIKNEGDKPKLDF"
CDS 4091..4111
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
polyA_signal 4211..4259
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 4461..4889
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 4903..5232
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 5083..5218
/label=SV40 ori
/note="SV40 origin of replication"
CDS 5299..6093
/codon_start=1
/gene="aph(3')-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/label=Neo
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
polyA_signal 6269..6390
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
rep_origin complement(6841..7429)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(7600..8460)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(8461..8565)
/gene="bla"
/label=AmpR promoter
promoter complement(8929..9169)
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA"
ORIGIN
1 atgctagttg gcgcgttgac attgattatt gactagttat taatagtaat caattacggg
61 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
121
