pmU6-gRNA Catalog No. PVT10988
Packing 2ug
pmU6-gRNA Information
Function Mammal Editing plasmids
Promoter: mU6
Prokaryotic resistance: kanamycin Kan
Screening markers: Mycophenolate G418
Cloned strain: Escherichia coli DH5 alpha
Culture conditions: 37 degrees centigrade
pmU6-gRNA Description
PmU6-gRNA is a gRNA expression plasmid of mammalian cells.
pmU6-gRNA Sequence
LOCUS Exported 3542 bp ds-DNA circular SYN 20-DEC-2017
DEFINITION synthetic circular DNA
FEATURES Location/Qualifiers
source 1..3542
/organism="synthetic DNA construct"
/mol_type="other DNA"
rep_origin complement(3..458)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 670..984
/label=mU6 promoter
/note="RNA polymerase III promoter for mouse U6 snRNA (Das
et al., 1988)"
misc_RNA 1001..1076
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
rep_origin complement(1562..2150)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(2227..3021)
/codon_start=1
/gene="aph(3')-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/label=NeoR/KanR
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
promoter complement(3056..3413)
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 3070..3205
/label=SV40 ori
/note="SV40 origin of replication"
promoter complement(3415..3519)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
601 taaaacgacg gccagtgagc gcgcgtaata cgactcacta tagggcgaat tgggtacccg
661 tctcagatag atccgacgcc gccatctcta ggcccgcgcc ggccccctcg cacagacttg
721 tgggagaagc tcggctactc ccctgccccg gttaatttgc atataatatt tcctagtaac
781 tatagaggct taatgtgcga taaaagacag ataatctgtt ctttttaata ctagctacat
841 tttacatgat aggcttggat ttctataaga gatacaaata ctaaattatt attttaaaaa
901 acagcacaaa aggaaactca ccctaactgt aaagtaattg tgtgttttga gactataaat
961 atcccttgga gaaaagcctt gtttgtcttc gagaagacct gttttagagc tagaaatagc
1021 aagttaaaat aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt cggtgctttt
1081 tttgtactga gacgccgcgg tggagctcca gcttttgttc cctttagtga gggttaattg
1141 cgcgcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
1201 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
1261 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
1321 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
1381 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
1441 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
1501 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
1561 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
1621 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
1681 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
1741 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
1801 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
1861 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
1921 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
1981 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta
2041 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
2101 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
2161 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
2221 tcatgatcag aagaactcgt caagaaggcg atagaaggcg atgcgctgcg aatcgggagc
2281 ggcgataccg taaagcacga ggaagcggtc agcccattcg ccgccaagct cttcagcaat
2341 atcacgggta gccaacgcta tgtcctgata gcggtccgcc acacccagcc ggccacagtc
2401 gatgaatcca gaaaagcggc cattttccac catgatattc ggcaagcagg catcgccatg
2461 ggtcacgacg agatcctcgc cgtcgggcat gctcgccttg agcctggcga acagttcggc
2521 tggcgcgagc ccctgatgct cttcgtccag atcatcctga tcgacaagac cggcttccat
2581 ccgagtacgt gctcgctcga tgcgatgttt cgcttggtgg tcgaatgggc aggtagccgg
2641 atcaagcgta tgcagccgcc gcattgcatc agccatgatg gatactttct cggcaggagc
2701 aaggtgagat gacaggagat cctgccccgg cacttcgccc aatagcagcc agtcccttcc
2761 cgcttcagtg acaacgtcga gcacagctgc gcaaggaacg cccgtcgtgg ccagccacga
2821 tagccgcgct gcctcgtctt gcagttcatt cagggcaccg gacaggtcgg tcttgacaaa
2881 aagaaccggg cgcccctgcg ctgacagccg gaacacggcg gcatcagagc agccgattgt
2941 ctgttgtgcc cagtcatagc cgaatagcct ctccacccaa gcggccggag aacctgcgtg
3001 caatccatct tgttcaatca tgcgaaacga tcctcatcct gtctcttgat cgatctttgc
3061 aaaagcctag gcctccaaaa aagcctcctc actacttctg gaatagctca gaggccgagg
3121 cggcctcggc ctctgcataa ataaaaaaaa ttagtcagcc atggggcgga gaatgggcgg
3181 aactgggcgg agttaggggc gggatgggcg gagttagggg cgggactatg gttgctgact
3241 aattgagatg catgctttgc atacttctgc ctgctgggga gcctggggac tttccacacc
3301 tggttgctga ctaattgaga tgcatgcttt gcatacttct gcctgctggg gagcctgggg
3361 actttccaca ccctaactga cacacattcc acagctggtt ctttccgcct caggactctt
3421 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
3481 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
3541 ac
//
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