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基本信息
別名: | pgRNA-humanized;U6-gRNA |
體類型: | CRISPR/Cas系統 |
載體抗性: | 氨芐青霉素 |
載體標簽: | / |
篩選標記: | 新霉素/G418 |
啟動子: | sp6 |
表達水平: | 高拷貝 |
載體大小: | 3915 bp |
克隆方法: | TOPO Cloning |
5' 測序引物: | T7 |
3' 測序引物: | / |
組成型: | 組成型 |
備注 | An empty gRNA expression vector, used to create a gRNA to a specific sequence. |
質粒圖譜和多克隆位點信息
注意:
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載體描述
An empty gRNA expression vector, used to create a gRNA to a specific sequence.
載體序列
LOCUS Exported 3915 bp ds-DNA circular SYN 10-APR-2019
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pgRNA-humanized
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3915)
AUTHORS BT-Lab
TITLE Direct Submission
JOURNAL Exported Wednesday, Apr 10, 2019 from SnapGene Viewer 4.3.6
COMMENT Alias: gRNA_Cloning Vector,U6-gRNA
FEATURES Location/Qualifiers
source 1..3915
/organism="synthetic DNA construct"
/mol_type="other DNA"
protein_bind 107..128
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 143..173
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 181..197
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
primer_bind 205..221
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
promoter 239..257
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
protein_bind 342..366
/gene="mutant version of attB"
/label=attB2
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
promoter complement(803..821)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(828..844)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
CDS 982..1284
/codon_start=1
/gene="ccdB"
/product="CcdB, a bacterial toxin that poisons DNA gyrase"
/label=ccdB
/note="Plasmids containing the ccdB gene cannot be
propagated in standard E. coli strains."
/translation="QFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDKV
SRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
CDS 1633..2427
/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"
regulatory 2628..2637
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 2634..3008
/codon_start=1
/gene="Sh ble from Streptoalloteichus hindustanus"
/product="antibiotic-binding protein"
/label=BleoR
/note="confers resistance to bleomycin, phleomycin, and
Zeocin(TM)"
/translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
EFALRDPAGNCVHFVAEEQD"
rep_origin 3146..3734
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
ORIGIN
1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagctat
241 ttaggtgaca ctatagaata ctcaagctat gcatcaagct tggtaccgag ctcggatcca
301 ctagtaacgg ccgccagtgt gctggaattc gccctttaat gccaactttg tacaagaaag
361 ctgggtctag aaaaaaagca ccgactcggt gccacttttt caagttgata acggactagc
421 cttattttaa cttaagatat ataaagccaa gaaatcgaaa tactttcaag ttacggtaag
481 catatgatag tccattttaa aacataattt taaaactgca aactacccaa gaaattatta
541 ctttctacgt cacgtatttt gtactaatat ctttgtgttt acagtcaaat taattctaat
601 tatctctcta acagccttgt atcgtatatg caaatatgaa ggaatcatgg gaaataggcc
661 ctcttcctgc ccgaccttgg taccggatcc agtcgactga attggttcct ttaaagcctg
721 cttttttgta caaagggcga attctgcaga tatccatcac actggcggcc gctcgagcat
781 gcatctagag ggcccaattc gccctatagt gagtcgtatt acaattcact ggccgtcgtt
841 ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat
901 ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag
961 ttgcgcagcc tatacgtacg gcagtttaag gtttacacct ataaaagaga gagccgttat
1021 cgtctgtttg tggatgtaca gagtgatatt attgacacgc cggggcgacg gatggtgatc
1081 cccctggcca gtgcacgtct gctgtcagat aaagtctccc gtgaacttta cccggtggtg
1141 catatcgggg atgaaagctg gcgcatgatg accaccgata tggccagtgt gccggtctcc
1201 gttatcgggg aagaagtggc tgatctcagc caccgcgaaa atgacatcaa aaacgccatt
1261 aacctgatgt tctggggaat ataaatgtca ggcatgagat tatcaaaaag gatcttcacc
1321 tagatccttt tcacgtagaa agccagtccg cagaaacggt gctgaccccg gatgaatgtc
1381 agctactggg ctatctggac aagggaaaac gcaagcgcaa agagaaagca ggtagcttgc
1441 agtgggctta catggcgata gctagactgg gcggttttat ggacagcaag cgaaccggaa
1501 ttgccagctg gggcgccctc tggtaaggtt gggaagccct gcaaagtaaa ctggatggct
1561 ttctcgccgc caaggatctg atggcgcagg ggatcaagct ctgatcaaga gacaggatga
1621 ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
1681 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
1741 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
1801 ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
1861 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
1921 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
1981 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
2041 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
2101 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
2161 agcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
2221 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
2281 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
2341 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
2401 tatcgccttc ttgacgagtt cttctgaatt attaacgctt acaatttcct gatgcggtat
2461 tttctcctta cgcatctgtg cggtatttca caccgcatac aggtggcact tttcggggaa
2521 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
2581 tgagacaata accctgataa atgcttcaat aatagcacgt gaggagggcc accatggcca
2641 agttgaccag tgccgttccg gtgctcaccg cgcgcgacgt cgccggagcg gtcgagttct
2701 ggaccgaccg gctcgggttc tcccgggact tcgtggagga cgacttcgcc ggtgtggtcc
2761 gggacgacgt gaccctgttc atcagcgcgg tccaggacca ggtggtgccg gacaacaccc
2821 tggcctgggt gtgggtgcgc ggcctggacg agctgtacgc cgagtggtcg gaggtcgtgt
2881 ccacgaactt ccgggacgcc tccgggccgg ccatgaccga gatcggcgag cagccgtggg
2941 ggcgggagtt cgccctgcgc gacccggccg gcaactgcgt gcacttcgtg gccgaggagc
3001 aggactgaca cgtgctaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
3061 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
3121 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
3181 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
3241 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag
3301 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
3361 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
3421 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
3481 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
3541 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
3601 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
3661 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
3721 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctgggc ttttgctggc
3781 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
3841 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
3901 gcgaggaagc ggaag
//