pUC57-sgRNA
PVT10920 2ug
pUC57-sgRNA Description
The pUC57-sgRNA plasmid is a vector for sgRNA transcription in vitro. The gRNA fragment can be digested by BsaI and transcribed by T7 promoter.
Promoter: T7 promoter
Replicon: pUC ori
Plasmid size: 2792bp
Prokaryotic resistance: kanamycin Kan
Clone strain: Escherichia coli DH5 alpha
Culture conditions: 37 C, aerobic, LB
pUC57-sgRNA Reference
Efficient genome modification by CRISPR-Cas9 nickase with minimal off-target effects.
pUC57-sgRNA Sequence
LOCUS Exported 2792 bp ds-DNA circular SYN 26-OCT-2017
DEFINITION synthetic circular DNA
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 2792)
AUTHORS .
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..2792
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 442..460
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
misc_RNA 482..557
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
rep_origin complement(1018..1606)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(1777..2592)
/codon_start=1
/gene="aph(3')-Ia"
/product="aminoglycoside phosphotransferase"
/label=KanR
/note="confers resistance to kanamycin in bacteria or G418
(Geneticin(R)) in eukaryotes"
/translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
promoter complement(2593..2697)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
361 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt cgagctcggt acctcgcgaa
421 tgcatctaga tatcggatcc ctaatacgac tcactatagg tgagaccgag agagggtctc
481 agttttagag ctagaaatag caagttaaaa taaggctagt ccgttatcaa cttgaaaaag
541 tggcaccgag tcggtgcttt ttttaaaggg cccgtcgact gcagaggcct gcatgcaagc
601 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
661 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
721 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
781 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgcggccgc
841 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
901 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
961 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
1021 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
1081 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
1141 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
1201 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
1261 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
1321 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
1381 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
1441 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
1501 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
1561 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
1621 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
1681 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
1741 aatctaaagt atatatgagt aaacttggtc tgacagttag aaaaactcat cgagcatcaa
1801 atgaaactgc aatttattca tatcaggatt atcaatacca tatttttgaa aaagccgttt
1861 ctgtaatgaa ggagaaaact caccgaggca gttccatagg atggcaagat cctggtatcg
1921 gtctgcgatt ccgactcgtc caacatcaat acaacctatt aatttcccct cgtcaaaaat
1981 aaggttatca agtgagaaat caccatgagt gacgactgaa tccggtgaga atggcaaaag
2041 tttatgcatt tctttccaga cttgttcaac aggccagcca ttacgctcgt catcaaaatc
2101 actcgcatca accaaaccgt tattcattcg tgattgcgcc tgagcgagac gaaatacgcg
2161 atcgctgtta aaaggacaat tacaaacagg aatcgaatgc aaccggcgca ggaacactgc
2221 cagcgcatca acaatatttt cacctgaatc aggatattct tctaatacct ggaatgctgt
2281 tttcccaggg atcgcagtgg tgagtaacca tgcatcatca ggagtacgga taaaatgctt
2341 gatggtcgga agaggcataa attccgtcag ccagtttagt ctgaccatct catctgtaac
2401 atcattggca acgctacctt tgccatgttt cagaaacaac tctggcgcat cgggcttccc
2461 atacaatcga tagattgtcg cacctgattg cccgacatta tcgcgagccc atttataccc
2521 atataaatca gcatccatgt tggaatttaa tcgcggccta gagcaagacg tttcccgttg
2581 aatatggctc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
2641 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
2701 atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta
2761 taaaaatagg cgtatcacga ggccctttcg tc
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文献和实验
技术资料