lentiSAMv2
Catalog No. PVT10983
Packing 2ug
lentiSAMv2 Information
Function Mammal Editing plasmids
Prokaryotic resistance: ampicillin Amp
Screening marker: bleomycin Zeo, magnaporin Blast
Cloned strain: Escherichia coli Stbl3
Culture conditions: 37 degrees centigrade
lentiSAMv2 Description
LentiSAMv2 is a mammalian CAS9 and gRNA expression plasmid
lentiSAMv2 Sequence
LOCUS Exported 13500 bp ds-DNA circular SYN 04-DEC-2017
DEFINITION synthetic circular DNA
FEATURES Location/Qualifiers
source 1..13500
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 118..447
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 298..433
/label=SV40 ori
/note="SV40 origin of replication"
promoter 495..542
/label=EM7 promoter
/note="synthetic bacterial promoter "
CDS 561..935
/codon_start=1
/gene="Sh ble from Streptoalloteichus hindustanus"
/product="antibiotic-binding protein"
/label=Zeo
/note="confers resistance to bleomycin, phleomycin, and
Zeocin(TM)"
/translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
EFALRDPAGNCVHFVAEEQD"
polyA_signal 1065..1186
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
rep_origin complement(1637..2225)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(2396..3256)
/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(3257..3361)
/gene="bla"
/label=AmpR promoter
enhancer 3627..4006
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 4008..4206
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
LTR 4224..4404
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 4451..4576
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 5069..5302
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
misc_feature 5829..5946
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 5996..6236
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA"
misc_RNA 6282..6300
/label=MS2 stem loop
/note="stem loop that binds the bacteriophage MS2 coat
protein"
misc_RNA 6352..6370
/label=MS2 stem loop
/note="stem loop that binds the bacteriophage MS2 coat
protein"
promoter 6457..6668
/label=EF-1-alpha core promoter
/note="core promoter for human elongation factor
EF-1-alpha"
CDS 6685..6732
/codon_start=1
/product="bipartite nuclear localization signal from
nucleoplasmin"
/label=nucleoplasmin NLS
/translation="KRPAATKKAGQAKKKK"
CDS 6733..10833
/codon_start=1
/product="catalytically dead mutant of the Cas9
endonuclease from the Streptococcus pyogenes Type II
CRISPR/Cas system"
/label=dCas9
/note="RNA-guided DNA-binding protein that lacks
endonuclease activity due to the D10A mutation in the RuvC
catalytic domain and the N863A mutation in the HNH
catalytic domain"
/translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF