| 质粒名称: | pAC149-pCR8-dCas9VP160 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 48221 |
| 存储实验室: | Rudolf Jaenisch |
|
载体骨架基本信息 |
|
| 载体名称: | pCR8/GW/TOPO |
| 载体出品公司: | Invitrogen |
| 载体抗性: | 大观霉素 |
| 空载体大小: | 2799 bp |
| 完整质粒大小: | 7442 bp |
| 载体类型: | CRISPR,网关的供体 |
| 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: | dCas9(D10A;H840A) fusion with VP160 activation domain |
| 插入大小: | 4643 bp |
| 基因突变: | D10A;H840A nuclease-deficient |
| 标签/融合蛋白: | HA Tag（N端） VP160（C端） |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | EcoRI(未破坏) |
| 3' 克隆位点: | EcoRI(未破坏) |
| 5' 测序引物 | M13F |
| 3' 测序引物 | M13R |
|
文献引用方法 |
|
| 材料和方法部分: | pAC149-pCR8-dCas9VP160 was a gift from Rudolf Jaenisch (Addgene plasmid # 48221) |
| 参考文献部分: | Multiplexed activation of endogenous genes by CRISPR-on, an RNA-guided transcriptional activator system. Cheng AW, Wang H, Yang H, Shi L, Katz Y, Theunissen TW, Rangarajan S, Shivalila CS, Dadon DB, Jaenisch R. Cell Res. 2013 Aug 27. doi: 10.1038/cr.2013.122. 10.1038/cr.2013.122 PubMed 23979020 |
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 pAC149-pCR8-dCas9VP160 7442 bp DNA SYN
DEFINITION pAC149-pCR8-dCas9VP160
ACCESSION
KEYWORDS
SOURCE
ORGANISM other sequences; artificial sequences; vectors.
FEATURES Location/Qualifiers
source 1..7442
/organism="pAC149-pCR8-dCas9VP160"
/mol_type="other DNA"
terminator complement(268..295)
/label="rrnB_T2_terminator"
terminator complement(427..470)
/label="rrnB_T1_terminator"
misc_feature 522..544
/label="M13_pUC_fwd_primer"
promoter 537..553
/label="M13_forward20_primer"
misc_feature complement(569..668)
/label="attL1"
CDS 628..1080
/label="ORF frame 1"
CDS 641..5305
/label="ORF frame 2"
misc_feature 692..718
/label="HA"
misc_feature 722..742
/label="SV40 NLS"
misc_feature 728..742
/label="NLS"
misc_feature 5321..5420
/label="attL2"
misc_feature 5333..5420
/label="attL1"
promoter complement(5438..5456)
/label="T7_promoter"
promoter complement(5457..5475)
/label="M13_reverse_primer"
gene 5689..6699
/label="spect"
/gene="spect"
CDS 5689..6699
/label="ORF frame 1"
rep_origin 6776..7395
/label="pBR322_origin"
ORIGIN
1 CTTTCCTGCG TTATCCCCTG ATTCTGTGGA TAACCGTATT ACCGCCTTTG AGTGAGCTGA
61 TACCGCTCGC CGCAGCCGAA CGACCGAGCG CAGCGAGTCA GTGAGCGAGG AAGCGGAAGA
121 GCGCCCAATA CGCAAACCGC CTCTCCCCGC GCGTTGGCCG ATTCATTAAT GCAGCTGGCA
181 CGACAGGTTT CCCGACTGGA AAGCGGGCAG TGAGCGCAAC GCAATTAATA CGCGTACCGC
241 TAGCCAGGAA GAGTTTGTAG AAACGCAAAA AGGCCATCCG TCAGGATGGC CTTCTGCTTA
301 GTTTGATGCC TGGCAGTTTA TGGCGGGCGT CCTGCCCGCC ACCCTCCGGG CCGTTGCTTC
361 ACAACGTTCA AATCCGCTCC CGGCGGATTT GTCCTACTCA GGAGAGCGTT CACCGACAAA
421 CAACAGATAA AACGAAAGGC CCAGTCTTCC GACTGAGCCT TTCGTTTTAT TTGATGCCTG
481 GCAGTTCCCT ACTCTCGCGT TAACGCTAGC ATGGATGTTT TCCCAGTCAC GACGTTGTAA
541 AACGACGGCC AGTCTTAAGC TCGGGCCCCA AATAATGATT TTATTTTGAC TGATAGTGAC
601 CTGTTCGTTG CAACAAATTG ATGAGCAATG CTTTTTTATA ATGCCAACTT TGTACAAAAA
661 AGCAGGCTCC GAATTcACCG GTgccaccAT GTACCCATAC GATGTTCCAG ATTACGCTTC
721 GCCGAAGAAA AAGCGCAAGG TCGAAGCGTC CGACAAGAAG TACAGCATCG GCCTGGCCAT
781 CGGCACCAAC TCTGTGGGCT GGGCCGTGAT CACCGACGAG TACAAGGTGC CCAGCAAGAA
841 ATTCAAGGTG CTGGGCAACA CCGACCGGCA CAGCATCAAG AAGAACCTGA TCGGAGCCCT
901 GCTGTTCGAC AGCGGCGAAA CAGCCGAGGC CACCCGGCTG AAGAGAACCG CCAGAAGAAG
961 ATACACCAGA CGGAAGAACC GGATCTGCTA TCTGCAAGAG ATCTTCAGCA ACGAGATGGC
1021 CAAGGTGGAC GACAGCTTCT TCCACAGACT GGAAGAGTCC TTCCTGGTGG AAGAGGATAA
1081 GAAGCACGAG CGGCACCCCA TCTTCGGCAA CATCGTGGAC GAGGTGGCCT ACCACGAGAA
1141 GTACCCCACC ATCTACCACC TGAGAAAGAA ACTGGTGGAC AGCACCGACA AGGCCGACCT
1201 GCGGCTGATC TATCTGGCCC TGGCCCACAT GATCAAGTTC CGGGGCCACT TCCTGATCGA
1261 GGGCGACCTG AACCCCGACA ACAGCGACGT GGACAAGCTG TTCATCCAGC TGGTGCAGAC
1321 CTACAACCAG CTGTTCGAGG AAAACCCCAT CAACGCCAGC GGCGTGGACG CCAAGGCCAT
1381 CCTGTCTGCC AGACTGAGCA AGAGCAGACG GCTGGAAAAT CTGATCGCCC AGCTGCCCGG
1441 CGAGAAGAAG AATGGCCTGT TCGGCAACCT GATTGCCCTG AGCCTGGGCC TGACCCCCAA
1501 CTTCAAGAGC AACTTCGACC TGGCCGAGGA TGCCAAACTG CAGCTGAGCA AGGACACCTA
1561 CGACGACGAC CTGGACAACC TGCTGGCCCA GATCGGCGAC CAGTACGCCG ACCTGTTTCT
1621 GGCCGCCAAG AACCTGTCCG ACGCCATCCT GCTGAGCGAC ATCCTGAGAG TGAACACCGA
1681 GATCACCAAG GCCCCCCTGA GCGCCTCTAT GATCAAGAGA TACGACGAGC ACCACCAGGA
1741 CCTGACCCTG CTGAAAGCTC TCGTGCGGCA GCAGCTGCCT GAGAAGTACA AAGAGATTTT
1801 CTTCGACCAG AGCAAGAACG GCTACGCCGG CTACATTGAC GGCGGAGCCA GCCAGGAAGA
1861 GTTCTACAAG TTCATCAAGC CCATCCTGGA AAAGATGGAC GGCACCGAGG AACTGCTCGT
1921 GAAGCTGAAC AGAGAGGACC TGCTGCGGAA GCAGCGGACC TTCGACAACG GCAGCATCCC
1981 CCACCAGATC CACCTGGGAG AGCTGCACGC CATTCTGCGG CGGCAGGAAG ATTTTTACCC
2041 ATTCCTGAAG GACAACCGGG AAAAGATCGA GAAGATCCTG ACCTTCCGCA TCCCCTACTA
2101 CGTGGGCCCT CTGGCCAGGG GAAACAGCAG ATTCGCCTGG ATGACCAGAA AGAGCGAGGA
2161 AACCATCACC CCCTGGAACT TCGAGGAAGT GGTGGACAAG GGCGCTTCCG CCCAGAGCTT
2221 CATCGAGCGG ATGACCAACT TCGATAAGAA CCTGCCCAAC GAGAAGGTGC TGCCCAAGCA
2281 CAGCCTGCTG TACGAGTACT TCACCGTGTA TAACGAGCTG ACCAAAGTGA AATACGTGAC
2341 CGAGGGAATG AGAAAGCCCG CCTTCCTGAG CGGCGAGCAG AAAAAGGCCA TCGTGGACCT
2401 GCTGTTCAAG ACCAACCGGA AAGTGACCGT GAAGCAGCTG AAAGAGGACT ACTTCAAGAA
2461 AATCGAGTGC TTCGACTCCG TGGAAATCTC CGGCGTGGAA GATCGGTTCA ACGCCTCCCT
2521 GGGCACATAC CACGATCTGC TGAAAATTAT CAAGGACAAG GACTTCCTGG ACAATGAGGA
2581 AAACGAGGAC ATTCTGGAAG ATATCGTGCT GACCCTGACA CTGTTTGAGG ACAGAGAGAT
2641 GATCGAGGAA CGGCTGAAAA CCTATGCCCA CCTGTTCGAC GACAAAGTGA TGAAGCAGCT
2701 GAAGCGGCGG AGATACACCG GCTGGGGCAG GCTGAGCCGG AAGCTGATCA ACGGCATCCG
2761 GGACAAGCAG TCCGGCAAGA CAATCCTGGA TTTCCTGAAG TCCGACGGCT TCGCCAACAG
2821 AAACTTCATG CAGCTGATCC ACGACGACAG CCTGACCTTT AAAGAGGACA TCCAGAAAGC
2881 CCAGGTGTCC GGCCAGGGCG ATAGCCTGCA CGAGCACATT GCCAATCTGG CCGGCAGCCC
2941 CGCCATTAAG AAGGGCATCC TGCAGACAGT GAAGGTGGTG GACGAGCTCG TGAAAGTGAT
3001 GGGCCGGCAC AAGCCCGAGA ACATCGTGAT CGAAATGGCC AGAGAGAACC AGACCACCCA
3061 GAAGGGACAG AAGAACAGCC GCGAGAGAAT GAAGCGGATC GAAGAGGGCA TCAAAGAGCT
3121 GGGCAGCCAG ATCCTGAAAG AACACCCCGT GGAAAACACC CAGCTGCAGA ACGAGAAGCT
3181 GTACCTGTAC TACCTGCAGA ATGGGCGGGA TATGTACGTG GACCAGGAAC TGGACATCAA
3241 CCGGCTGTCC GACTACGATG TGGACGCCAT CGTGCCTCAG AGCTTTCTGA AGGACGACTC
3301 CATCGACAAC AAGGTGCTGA CCAGAAGCGA CAAGAACCGG GGCAAGAGCG ACAACGTGCC
3361 CTCCGAAGAG GTCGTGAAGA AGATGAAGAA CTACTGGCGG CAGCTGCTGA ACGCCAAGCT
3421 GATTACCCAG AGAAAGTTCG ACAATCTGAC CAAGGCCGAG AGAGGCGGCC TGAGCGAACT
3481 GGATAAGGCC GGCTTCATCA AGAGACAGCT GGTGGAAACC CGGCAGATCA CAAAGCACGT
3541 GGCACAGATC CTGGACTCCC GGATGAACAC TAAGTACGAC GAGAATGACA AGCTGATCCG
3601 GGAAGTGAAA GTGATCACCC TGAAGTCCAA GCTGGTGTCC GATTTCCGGA AGGATTTCCA
3661 GTTTTACAAA GTGCGCGAGA TCAACAACTA CCACCACGCC CACGACGCCT ACCTGAACGC
3721 CGTCGTGGGA ACCGCCCTGA TCAAAAAGTA CCCTAAGCTG GAAAGCGAGT TCGTGTACGG
3781 CGACTACAAG GTGTACGACG TGCGGAAGAT GATCGCCAAG AGCGAGCAGG AAATCGGCAA
3841 GGCTACCGCC AAGTACTTCT TCTACAGCAA CATCATGAAC TTTTTCAAGA CCGAGATTAC
3901 CCTGGCCAAC GGCGAGATCC GGAAGCGGCC TCTGATCGAG ACAAACGGCG AAACCGGGGA
3961 GATCGTGTGG GATAAGGGCC GGGATTTTGC CACCGTGCGG AAAGTGCTGA GCATGCCCCA
4021 AGTGAATATC GTGAAAAAGA CCGAGGTGCA GACAGGCGGC TTCAGCAAAG AGTCTATCCT
4081 GCCCAAGAGG AACAGCGATA AGCTGATCGC CAGAAAGAAG GACTGGGACC CTAAGAAGTA
4141 CGGCGGCTTC GACAGCCCCA CCGTGGCCTA TTCTGTGCTG GTGGTGGCCA AAGTGGAAAA
4201 GGGCAAGTCC AAGAAACTGA AGAGTGTGAA AGAGCTGCTG GGGATCACCA TCATGGAAAG
4261 AAGCAGCTTC GAGAAGAATC CCATCGACTT TCTGGAAGCC AAGGGCTACA AAGAAGTGAA
4321 AAAGGACCTG ATCATCAAGC TGCCTAAGTA CTCCCTGTTC GAGCTGGAAA ACGGCCGGAA
4381 GAGAATGCTG GCCTCTGCCG GCGAACTGCA GAAGGGAAAC GAACTGGCCC TGCCCTCCAA
4441 ATATGTGAAC TTCCTGTACC TGGCCAGCCA CTATGAGAAG CTGAAGGGCT CCCCCGAGGA
4501 TAATGAGCAG AAACAGCTGT TTGTGGAACA GCACAAGCAC TACCTGGACG AGATCATCGA
4561 GCAGATCAGC GAGTTCTCCA AGAGAGTGAT CCTGGCCGAC GCTAATCTGG ACAAAGTGCT
4621 GTCCGCCTAC AACAAGCACC GGGATAAGCC CATCAGAGAG CAGGCCGAGA ATATCATCCA
4681 CCTGTTTACC CTGACCAATC TGGGAGCCCC TGCCGCCTTC AAGTACTTTG ACACCACCAT
4741 CGACCGGAAG AGGTACACCA GCACCAAAGA GGTGCTGGAC GCCACCCTGA TCCACCAGAG
4801 CATCACCGGC CTGTACGAGA CACGGATCGA CCTGTCTCAG CTGGGAGGCG ACAGCCCCAA
4861 GAAGAAGAGA AAGGTGGAGG CCAGCgggcc ggccGGATCC GGGCGCGCCG ACGCGCTGGA
4921 CGATTTCGAT CTCGACATGC TGGGTTCTGA TGCCCTCGAT GACTTTGACC TGGATATGTT
4981 GGGAAGCGAC GCATTGGATG ACTTTGATCT GGACATGCTC GGCTCCGATG CTCTGGACGA
5041 TTTCGATCTC GATATGTTAg ggtcagacgc actggatgat ttcgaccttg atatgttgGG
5101 AAGCGATGCC CTTGATGATT TCGACCTGGA CATGCTCggc agcgacgccc tggacgattt
5161 cgatctggac atgctgGGGT CCGATGCCTT GGATGATTTT GACTTGGATA TGCTGgggag
5221 tgatgccctg gacgactttg acctggacat gctgGGCTCC GATGCGCTCG ATGACTTCGA
5281 TTTGGAtATG TTGTATATCG ATtgaTTAAT TAAGAATTCG ACCCAGCTTT CTTGTACAAA
5341 GTTGGCATTA TAAAAAATAA TTGCTCATCA ATTTGTTGCA ACGAACAGGT CACTATCAGT
5401 CAAAATAAAA TCATTATTTG CCATCCAGCT GATATCCCCT ATAGTGAGTC GTATTACATG
5461 GTCATAGCTG TTTCCTGGCA GCTCTGGCCC GTGTCTCAAA ATCTCTGATG TTACATTGCA
5521 CAAGATAAAA ATATATCATC ATGCCTCCTC TAGACCAGCC AGGACAGAAA TGCCTCGACT
5581 TCGCTGCTGC CCAAGGTTGC CGGGTGACGC ACACCGTGGA AACGGATGAA GGCACGAACC
5641 CAGTGGACAT AAGCCTGTTC GGTTCGTAAG CTGTAATGCA AGTAGCGTAT GCGCTCACGC
5701 AACTGGTCCA GAACCTTGAC CGAACGCAGC GGTGGTAACG GCGCAGTGGC GGTTTTCATG
5761 GCTTGTTATG ACTGTTTTTT TGGGGTACAG TCTATGCCTC GGGCATCCAA GCAGCAAGCG
5821 CGTTACGCCG TGGGTCGATG TTTGATGTTA TGGAGCAGCA ACGATGTTAC GCAGCAGGGC
5881 AGTCGCCCTA AAACAAAGTT AAACATCATG AGGGAAGCGG TGATCGCCGA AGTATCGACT
5941 CAACTATCAG AGGTAGTTGG CGTCATCGAG CGCCATCTCG AACCGACGTT GCTGGCCGTA
6001 CATTTGTACG GCTCCGCAGT GGATGGCGGC CTGAAGCCAC ACAGTGATAT TGATTTGCTG
6061 GTTACGGTGA CCGTAAGGCT TGATGAAACA ACGCGGCGAG CTTTGATCAA CGACCTTTTG
6121 GAAACTTCGG CTTCCCCTGG AGAGAGCGAG ATTCTCCGCG CTGTAGAAGT CACCATTGTT
6181 GTGCACGACG ACATCATTCC GTGGCGTTAT CCAGCTAAGC GCGAACTGCA ATTTGGAGAA
6241 TGGCAGCGCA ATGACATTCT TGCAGGTATC TTCGAGCCAG CCACGATCGA CATTGATCTG
6301 GCTATCTTGC TGACAAAAGC AAGAGAACAT AGCGTTGCCT TGGTAGGTCC AGCGGCGGAG
6361 GAACTCTTTG ATCCGGTTCC TGAACAGGAT CTATTTGAGG CGCTAAATGA AACCTTAACG
6421 CTATGGAACT CGCCGCCCGA CTGGGCTGGC GATGAGCGAA ATGTAGTGCT TACGTTGTCC
6481 CGCATTTGGT ACAGCGCAGT AACCGGCAAA ATCGCGCCGA AGGATGTCGC TGCCGACTGG
6541 GCAATGGAGC GCCTGCCGGC CCAGTATCAG CCCGTCATAC TTGAAGCTAG ACAGGCTTAT
6601 CTTGGACAAG AAGAAGATCG CTTGGCCTCG CGCGCAGATC AGTTGGAAGA ATTTGTCCAC
6661 TACGTGAAAG GCGAGATCAC CAAGGTAGTC GGCAAATAAC CCTCGAGCCA CCCATGACCA
6721 AAATCCCTTA ACGTGAGTTA CGCGTCGTTC CACTGAGCGT CAGACCCCGT AGAAAAGATC
6781 AAAGGATCTT CTTGAGATCC TTTTTTTCTG CGCGTAATCT GCTGCTTGCA AACAAAAAAA
6841 CCACCGCTAC CAGCGGTGGT TTGTTTGCCG GATCAAGAGC TACCAACTCT TTTTCCGAAG
6901 GTAACTGGCT TCAGCAGAGC GCAGATACCA AATACTGTCC TTCTAGTGTA GCCGTAGTTA
6961 GGCCACCACT TCAAGAACTC TGTAGCACCG CCTACATACC TCGCTCTGCT AATCCTGTTA
7021 CCAGTGGCTG CTGCCAGTGG CGATAAGTCG TGTCTTACCG GGTTGGACTC AAGACGATAG
7081 TTACCGGATA AGGCGCAGCG GTCGGGCTGA ACGGGGGGTT CGTGCACACA GCCCAGCTTG
7141 GAGCGAACGA CCTACACCGA ACTGAGATAC CTACAGCGTG AGCATTGAGA AAGCGCCACG
7201 CTTCCCGAAG GGAGAAAGGC GGACAGGTAT CCGGTAAGCG GCAGGGTCGG AACAGGAGAG
7261 CGCACGAGGG AGCTTCCAGG GGGAAACGCC TGGTATCTTT ATAGTCCTGT CGGGTTTCGC
7321 CACCTCTGAC TTGAGCGTCG ATTTTTGTGA TGCTCGTCAG GGGGGCGGAG CCTATGGAAA
7381 AACGCCAGCA ACGCGGCCTT TTTACGGTTC CTGGCCTTTT GCTGGCCTTT TGCTCACATG
7441 TT
//
