| 质粒名称: | pLX-sgRNA |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 50662 |
| 存储实验室: | Eric Lander, David Sabatini |
|
载体骨架基本信息 |
|
| 载体名称: | pLX304 |
| 载体抗性: | 氨苄霉素 |
| 空载体大小: | 7000 bp |
| 完整质粒大小: | 7500 bp |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | 稻瘟 |
| 高拷贝/低拷贝: | 未知 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | sgAAVS1 |
| 插入大小: | 450 bp |
| 启动子 | U6 |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | XhoI(未破坏) |
| 3' 克隆位点: | NheI(未破坏) |
| 5' 测序引物 | cgggtttattacagggacagcag |
| 3' 测序引物 | taccagtcaatctttcacaaattttgt |
|
文献引用方法 |
|
| 材料和方法部分: | pLX-sgRNA was a gift from Eric Lander & David Sabatini (Addgene plasmid # 50662) |
| 参考文献部分: | Genetic screens in human cells using the CRISPR-Cas9 system. Wang T, Wei JJ, Sabatini DM, Lander ES. Science. 2014 Jan 3;343(6166):80-4. doi: 10.1126/science.1246981. Epub 2013 Dec 12. 10.1126/science.1246981 PubMed 24336569 |
买家导航
载体基本信息
载体图谱质粒图谱和多克隆位点信息
载体简介
载体序列
LOCUS pLX-sgRNA 7477 bp DNA SYN
DEFINITION pLX-sgRNA
ACCESSION
KEYWORDS
SOURCE
ORGANISM other sequences; artificial sequences; vectors.
FEATURES Location/Qualifiers
source 1..7477
/organism="pLX-sgRNA"
/mol_type="other DNA"
misc_feature 232..412
/label="HIV-1_5_LTR"
misc_feature 232..412
/label="truncHIV-1_3_LTR"
misc_feature 523..567
/label="HIV-1_psi_pack"
CDS 955..1938
/label="ORF frame 1"
misc_feature 1077..1310
/label="RRE"
CDS complement(1683..2528)
/label="ORF frame 3"
gene 2226..2621
/label="blast"
/gene="blast"
misc_feature 2685..2700
/label="cPPT"
promoter 2901..3157
/label="hU6_promoter"
misc_feature 3087..3106
/label="LKO1_5_primer"
misc_feature 3336..3911
/label="WPRE"
CDS 3424..3933
/label="ORF frame 1"
CDS 3437..3967
/label="ORF frame 2"
misc_feature 3977..3992
/label="cPPT"
misc_feature 3977..3998
/label="U3PPT"
misc_feature 3994..4046
/label="delta_U3"
misc_feature 4047..4227
/label="HIV-1_5_LTR"
misc_feature 4047..4227
/label="truncHIV-1_3_LTR"
terminator 4308..4427
/label="SV40_PA_terminator"
misc_feature 4396..4415
/label="EBV_rev_primer"
rep_origin 4445..4543
/label="SV40_origin"
misc_feature 4528..4547
/label="SV40pro_F_primer"
promoter complement(4622..4640)
/label="T7_promoter"
promoter complement(4650..4666)
/label="M13_forward20_primer"
misc_feature complement(4659..4681)
/label="M13_pUC_fwd_primer"
misc_feature 4659..4807
/label="lacZ_a"
rep_origin 4825..5131
/label="f1_origin"
promoter 5324..5352
/label="AmpR_promoter"
gene 5394..6254
/label="Ampicillin"
/gene="Ampicillin"
CDS 5394..6254
/label="ORF frame 3"
rep_origin 6409..7028
/label="pBR322_origin"
promoter 7337..7366
/label="lac_promoter"
misc_feature 7380..7402
/label="M13_pUC_rev_primer"
promoter 7401..7419
/label="M13_reverse_primer"
promoter 7436..7455
/label="T3_promoter"
ORIGIN
1 ttaatgtagt cttatgcaat actcttgtag tcttgcaaca tggtaacgat gagttagcaa
61 catgccttac aaggagagaa aaagcaccgt gcatgccgat tggtggaagt aaggtggtac
121 gatcgtgcct tattaggaag gcaacagacg ggtctgacat ggattggacg aaccactgaa
181 ttgccgcatt gcagagatat tgtatttaag tgcctagctc gatacataaa cgggtctctc
241 tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag
301 cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct
361 ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag cagtggcgcc
421 cgaacaggga cttgaaagcg aaagggaaac cagaggagct ctctcgacgc aggactcggc
481 ttgctgaagc gcgcacggca agaggcgagg ggcggcgact ggtgagtacg ccaaaaattt
541 tgactagcgg aggctagaag gagagagatg ggtgcgagag cgtcagtatt aagcggggga
601 gaattagatc gcgatgggaa aaaattcggt taaggccagg gggaaagaaa aaatataaat
661 taaaacatat agtatgggca agcagggagc tagaacgatt cgcagttaat cctggcctgt
721 tagaaacatc agaaggctgt agacaaatac tgggacagct acaaccatcc cttcagacag
781 gatcagaaga acttagatca ttatataata cagtagcaac cctctattgt gtgcatcaaa
841 ggatagagat aaaagacacc aaggaagctt tagacaagat agaggaagag caaaacaaaa
901 gtaagaccac cgcacagcaa gcggccgctg atcttcagac ctggaggagg agatatgagg
961 gacaattgga gaagtgaatt atataaatat aaagtagtaa aaattgaacc attaggagta
1021 gcacccacca aggcaaagag aagagtggtg cagagagaaa aaagagcagt gggaatagga
1081 gctttgttcc ttgggttctt gggagcagca ggaagcacta tgggcgcagc gtcaatgacg
1141 ctgacggtac aggccagaca attattgtct ggtatagtgc agcagcagaa caatttgctg
1201 agggctattg aggcgcaaca gcatctgttg caactcacag tctggggcat caagcagctc
1261 caggcaagaa tcctggctgt ggaaagatac ctaaaggatc aacagctcct ggggatttgg
1321 ggttgctctg gaaaactcat ttgcaccact gctgtgcctt ggaatgctag ttggagtaat
1381 aaatctctgg aacagatttg gaatcacacg acctggatgg agtgggacag agaaattaac
1441 aattacacaa gcttaataca ctccttaatt gaagaatcgc aaaaccagca agaaaagaat
1501 gaacaagaat tattggaatt agataaatgg gcaagtttgt ggaattggtt taacataaca
1561 aattggctgt ggtatataaa attattcata atgatagtag gaggcttggt aggtttaaga
1621 atagtttttg ctgtactttc tatagtgaat agagttaggc agggatattc accattatcg
1681 tttcagaccc acctcccaac cccgagggga cccttgcgcc ttttccaagg cagccctggg
1741 tttgcgcagg gacgcggctg ctctgggcgt ggttccggga aacgcagcgg cgccgaccct
1801 gggtctcgca cattcttcac gtccgttcgc agcgtcaccc ggatcttcgc cgctaccctt
1861 gtgggccccc cggcgacgct tcctgctccg cccctaagtc gggaaggttc cttgcggttc
1921 gcggcgtgcc ggacgtgaca aacggaagcc gcacgtctca ctagtaccct cgcagacgga
1981 cagcgccagg gagcaatggc agcgcgccga ccgcgatggg ctgtggccaa tagcggctgc
2041 tcagcagggc gcgccgagag cagcggccgg gaaggggcgg tgcgggaggc ggggtgtggg
2101 gcggtagtgt gggccctgtt cctgcccgcg cggtgttccg cattctgcaa gcctccggag
2161 cgcacgtcgg cagtcggctc cctcgttgac cgaatcaccg acctctctcc ccagggggta
2221 ccaccatggc caagcctttg tctcaagaag aatccaccct cattgaaaga gcaacggcta
2281 caatcaacag catccccatc tctgaagact acagcgtcgc cagcgcagct ctctctagcg
2341 acggccgcat cttcactggt gtcaatgtat atcattttac tgggggacct tgtgcagaac
2401 tcgtggtgct gggcactgct gctgctgcgg cagctggcaa cctgacttgt atcgtcgcga
2461 tcggaaatga gaacaggggc atcttgagcc cctgcggacg gtgccgacag gtgcttctcg
2521 atctgcatcc tgggatcaaa gccatagtga aggacagtga tggacagccg acggcagttg
2581 ggattcgtga attgctgccc tctggttatg tgtgggaggg cctgcagctg cagtagtaag
2641 aattctagat cttgagacaa atggcagtat tcatccacaa ttttaaaaga aaagggggga
2701 ttggggggta cagtgcaggg gaaagaatag tagacataat agcaacagac atacaaacta
2761 aagaattaca aaaacaaatt acaaaaattc aaaattttcg ggtttattac agggacagca
2821 gagatccact ttggcgccgg ctcgagTGTA CAAAAAAGCA GGCTTTAAAG GAACCAATTC
2881 AGTCGACTGG ATCCGGTACC AAGGTCGGGC AGGAAGAGGG CCTATTTCCC ATGATTCCTT
2941 CATATTTGCA TATACGATAC AAGGCTGTTA GAGAGATAAT TAGAATTAAT TTGACTGTAA
3001 ACACAAAGAT ATTAGTACAA AATACGTGAC GTAGAAAGTA ATAATTTCTT GGGTAGTTTG
3061 CAGTTTTAAA ATTATGTTTT AAAATGGACT ATCATATGCT TACCGTAACT TGAAAGTATT
3121 TCGATTTCTT GGCTTTATAT ATCTTGTGGA AAGGACGAAA CACCGGGGCC ACTAGGGACA
3181 GGATGTTTTA GAGCTAGAAA TAGCAAGTTA AAATAAGGCT AGTCCGTTAT CAACTTGAAA
3241 AAGTGGCACC GAGTCGGTGC TTTTTTTCTA GACCCAGCTT TCTTGTACAA AGTTGGCATT
3301 Agctagcgct aaccggtggc gcgttaagtc gacaatcaac ctctggatta caaaatttgt
3361 gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct
3421 ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat
3481 aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg
3541 gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag
3601 ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc
3661 tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg
3721 tcggggaaat catcgtcctt tccttggctg ctcgcctgtg ttgccacctg gattctgcgc
3781 gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc
3841 ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc
3901 tccctttggg ccgcctcccc gcgtcgactt taagaccaat gacttacaag gcagctgtag
3961 atcttagcca ctttttaaaa gaaaaggggg gactggaagg gctaattcac tcccaacgaa
4021 gacaagatct gctttttgct tgtactgggt ctctctggtt agaccagatc tgagcctggg
4081 agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc
4141 ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct
4201 tttagtcagt gtggaaaatc tctagcagta cgtatagtag ttcatgtcat cttattattc
4261 agtatttata acttgcaaag aaatgaatat cagagagtga gaggaacttg tttattgcag
4321 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
4381 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggctct
4441 agctatcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
4501 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc
4561 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctagggacgt acccaattcg
4621 ccctatagtg agtcgtatta cgcgcgctca ctggccgtcg ttttacaacg tcgtgactgg
4681 gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg
4741 cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc
4801 gaatgggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
4861 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
4921 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
4981 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
5041 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
5101 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
5161 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
5221 aaatttaacg cgaattttaa caaaatatta acgcttacaa tttaggtggc acttttcggg
5281 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
5341 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
5401 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
5461 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
5521 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
5581 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
5641 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
5701 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
5761 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
5821 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
5881 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
5941 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
6001 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
6061 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
6121 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
6181 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
6241 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
6301 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
6361 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
6421 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
6481 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
6541 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
6601 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
6661 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
6721 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
6781 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
6841 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
6901 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
6961 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
7021 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
7081 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
7141 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
7201 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca
7261 ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc
7321 attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga
7381 gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag cgcgcaatta
7441 accctcacta aagggaacaa aagctggagc tgcaagc
//
