| 质粒名称: | pY016 (pcDNA3.1-hLbCpf1) |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 69988 |
| 存储实验室: | Feng Zhang |
|
载体骨架基本信息 |
|
| 载体名称: | pcDNA3.1 |
| 载体抗性: | 氨苄霉素 |
| 空载体大小: | 5354 bp |
| 完整质粒大小: | 9227 bp |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | 新霉素(G418) |
| 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: | hLbCpf1 |
| 物种来源: | Lachnospiraceae_bacterium_ND2006 |
| 插入大小: | 3684 bp |
| 启动子 | CMV |
| 标签/融合蛋白: | NLS-3xHA(C端) |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | KpnI(未破坏) |
| 3' 克隆位点: | EcoRI(未破坏) |
| 5' 测序引物 | CMV-F |
| 3' 测序引物 | BGH-rev |
|
文献引用方法 |
|
| 材料和方法部分: | pY016 (pcDNA3.1-hLbCpf1) was a gift from Feng Zhang (Addgene plasmid # 69988) |
| 参考文献部分: | Cpf1 Is a Single RNA-Guided Endonuclease of a Class 2 CRISPR-Cas System. Zetsche B, Gootenberg JS, Abudayyeh OO, Slaymaker IM, Makarova KS, Essletzbichler P, Volz SE, Joung J, van der Oost J, Regev A, Koonin EV, Zhang F. Cell. 2015 Sep 23. pii: S0092-8674(15)01200-3. doi: 10.1016/j.cell.2015.09.038. 10.1016/j.cell.2015.09.038 PubMed 26422227 |
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pY016 (pcDNA3.1-hLbCpf1)载体质粒基本信息
pY016 (pcDNA3.1-hLbCpf1)质粒图谱载体图谱和pY016 (pcDNA3.1-hLbCpf1)载体序列质粒序列多克隆位点信息
质粒图谱.png)
pY016 (pcDNA3.1-hLbCpf1)质粒载体简介
pY016 (pcDNA3.1-hLbCpf1)质粒序列载体序列
LOCUS pY016 (pcDNA3.1-hLbCpf1) 9227 bp DNA SYN
DEFINITION pY016 (pcDNA3.1-hLbCpf1)
ACCESSION
KEYWORDS
SOURCE
ORGANISM other sequences; artificial sequences; vectors.
FEATURES Location/Qualifiers
source 1..9227
/organism="pY016 (pcDNA3.1-hLbCpf1)"
/mol_type="other DNA"
promoter 236..852
/label="CMV_immearly_promoter"
misc_feature 315..602
/label="CAG_enhancer"
misc_feature 769..789
/label="CMV_fwd_primer"
promoter 863..881
/label="T7_promoter"
CDS 929..4750
/label="ORF frame 2"
misc_feature 4667..4693
/label="HA"
misc_feature complement(4821..4838)
/label="BGH_rev_primer"
terminator 4824..5051
/label="bGH_PA_terminator"
rep_origin 5114..5420
/label="f1_origin"
misc_feature complement(5534..5554)
/label="pBABE_3_primer"
misc_feature complement(5540..5755)
/label="SV40_enhancer"
promoter 5552..5820
/label="SV40_promoter"
rep_origin 5719..5796
/label="SV40_origin"
misc_feature 5781..5800
/label="SV40pro_F_primer"
CDS 5935..6729
/label="ORF frame 1"
gene 5938..6726
/label="NeoR/KanR"
/gene="NeoR/KanR"
CDS complement(6244..6780)
/label="ORF frame 3"
terminator 6906..7025
/label="SV40_PA_terminator"
misc_feature 6994..7013
/label="EBV_rev_primer"
promoter complement(7069..7087)
/label="M13_reverse_primer"
misc_feature complement(7086..7108)
/label="M13_pUC_rev_primer"
promoter complement(7122..7151)
/label="lac_promoter"
gene complement(8231..9091)
/label="Ampicillin"
/gene="Ampicillin"
CDS complement(8231..9091)
/label="ORF frame 2"
promoter complement(9133..9161)
/label="AmpR_promoter"
ORIGIN
1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
901 gtttaaactt aagcttggta ccgccaccAT GAGCAAGCTG GAGAAGTTTA CAAACTGCTA
961 CTCCCTGTCT AAGACCCTGA GGTTCAAGGC CATCCCTGTG GGCAAGACCC AGGAGAACAT
1021 CGACAATAAG CGGCTGCTGG TGGAGGACGA GAAGAGAGCC GAGGATTATA AGGGCGTGAA
1081 GAAGCTGCTG GATCGCTACT ATCTGTCTTT TATCAACGAC GTGCTGCACA GCATCAAGCT
1141 GAAGAATCTG AACAATTACA TCAGCCTGTT CCGGAAGAAA ACCAGAACCG AGAAGGAGAA
1201 TAAGGAGCTG GAGAACCTGG AGATCAATCT GCGGAAGGAG ATCGCCAAGG CCTTCAAGGG
1261 CAACGAGGGC TACAAGTCCC TGTTTAAGAA GGATATCATC GAGACAATCC TGCCAGAGTT
1321 CCTGGACGAT AAGGACGAGA TCGCCCTGGT GAACAGCTTC AATGGCTTTA CCACAGCCTT
1381 CACCGGCTTC TTTGATAACA GAGAGAATAT GTTTTCCGAG GAGGCCAAGA GCACATCCAT
1441 CGCCTTCAGG TGTATCAACG AGAATCTGAC CCGCTACATC TCTAATATGG ACATCTTCGA
1501 GAAGGTGGAC GCCATCTTTG ATAAGCACGA GGTGCAGGAG ATCAAGGAGA AGATCCTGAA
1561 CAGCGACTAT GATGTGGAGG ATTTCTTTGA GGGCGAGTTC TTTAACTTTG TGCTGACACA
1621 GGAGGGCATC GACGTGTATA ACGCCATCAT CGGCGGCTTC GTGACCGAGA GCGGCGAGAA
1681 GATCAAGGGC CTGAACGAGT ACATCAACCT GTATAATCAG AAAACCAAGC AGAAGCTGCC
1741 TAAGTTTAAG CCACTGTATA AGCAGGTGCT GAGCGATCGG GAGTCTCTGA GCTTCTACGG
1801 CGAGGGCTAT ACATCCGATG AGGAGGTGCT GGAGGTGTTT AGAAACACCC TGAACAAGAA
1861 CAGCGAGATC TTCAGCTCCA TCAAGAAGCT GGAGAAGCTG TTCAAGAATT TTGACGAGTA
1921 CTCTAGCGCC GGCATCTTTG TGAAGAACGG CCCCGCCATC AGCACAATCT CCAAGGATAT
1981 CTTCGGCGAG TGGAACGTGA TCCGGGACAA GTGGAATGCC GAGTATGACG ATATCCACCT
2041 GAAGAAGAAG GCCGTGGTGA CCGAGAAGTA CGAGGACGAT CGGAGAAAGT CCTTCAAGAA
2101 GATCGGCTCC TTTTCTCTGG AGCAGCTGCA GGAGTACGCC GACGCCGATC TGTCTGTGGT
2161 GGAGAAGCTG AAGGAGATCA TCATCCAGAA GGTGGATGAG ATCTACAAGG TGTATGGCTC
2221 CTCTGAGAAG CTGTTCGACG CCGATTTTGT GCTGGAGAAG AGCCTGAAGA AGAACGACGC
2281 CGTGGTGGCC ATCATGAAGG ACCTGCTGGA TTCTGTGAAG AGCTTCGAGA ATTACATCAA
2341 GGCCTTCTTT GGCGAGGGCA AGGAGACAAA CAGGGACGAG TCCTTCTATG GCGATTTTGT
2401 GCTGGCCTAC GACATCCTGC TGAAGGTGGA CCACATCTAC GATGCCATCC GCAATTATGT
2461 GACCCAGAAG CCCTACTCTA AGGATAAGTT CAAGCTGTAT TTTCAGAACC CTCAGTTCAT
2521 GGGCGGCTGG GACAAGGATA AGGAGACAGA CTATCGGGCC ACCATCCTGA GATACGGCTC
2581 CAAGTACTAT CTGGCCATCA TGGATAAGAA GTACGCCAAG TGCCTGCAGA AGATCGACAA
2641 GGACGATGTG AACGGCAATT ACGAGAAGAT CAACTATAAG CTGCTGCCCG GCCCTAATAA
2701 GATGCTGCCA AAGGTGTTCT TTTCTAAGAA GTGGATGGCC TACTATAACC CCAGCGAGGA
2761 CATCCAGAAG ATCTACAAGA ATGGCACATT CAAGAAGGGC GATATGTTTA ACCTGAATGA
2821 CTGTCACAAG CTGATCGACT TCTTTAAGGA TAGCATCTCC CGGTATCCAA AGTGGTCCAA
2881 TGCCTACGAT TTCAACTTTT CTGAGACAGA GAAGTATAAG GACATCGCCG GCTTTTACAG
2941 AGAGGTGGAG GAGCAGGGCT ATAAGGTGAG CTTCGAGTCT GCCAGCAAGA AGGAGGTGGA
3001 TAAGCTGGTG GAGGAGGGCA AGCTGTATAT GTTCCAGATC TATAACAAGG ACTTTTCCGA
3061 TAAGTCTCAC GGCACACCCA ATCTGCACAC CATGTACTTC AAGCTGCTGT TTGACGAGAA
3121 CAATCACGGA CAGATCAGGC TGAGCGGAGG AGCAGAGCTG TTCATGAGGC GCGCCTCCCT
3181 GAAGAAGGAG GAGCTGGTGG TGCACCCAGC CAACTCCCCT ATCGCCAACA AGAATCCAGA
3241 TAATCCCAAG AAAACCACAA CCCTGTCCTA CGACGTGTAT AAGGATAAGA GGTTTTCTGA
3301 GGACCAGTAC GAGCTGCACA TCCCAATCGC CATCAATAAG TGCCCCAAGA ACATCTTCAA
3361 GATCAATACA GAGGTGCGCG TGCTGCTGAA GCACGACGAT AACCCCTATG TGATCGGCAT
3421 CGATAGGGGC GAGCGCAATC TGCTGTATAT CGTGGTGGTG GACGGCAAGG GCAACATCGT
3481 GGAGCAGTAT TCCCTGAACG AGATCATCAA CAACTTCAAC GGCATCAGGA TCAAGACAGA
3541 TTACCACTCT CTGCTGGACA AGAAGGAGAA GGAGAGGTTC GAGGCCCGCC AGAACTGGAC
3601 CTCCATCGAG AATATCAAGG AGCTGAAGGC CGGCTATATC TCTCAGGTGG TGCACAAGAT
3661 CTGCGAGCTG GTGGAGAAGT ACGATGCCGT GATCGCCCTG GAGGACCTGA ACTCTGGCTT
3721 TAAGAATAGC CGCGTGAAGG TGGAGAAGCA GGTGTATCAG AAGTTCGAGA AGATGCTGAT
3781 CGATAAGCTG AACTACATGG TGGACAAGAA GTCTAATCCT TGTGCAACAG GCGGCGCCCT
3841 GAAGGGCTAT CAGATCACCA ATAAGTTCGA GAGCTTTAAG TCCATGTCTA CCCAGAACGG
3901 CTTCATCTTT TACATCCCTG CCTGGCTGAC ATCCAAGATC GATCCATCTA CCGGCTTTGT
3961 GAACCTGCTG AAAACCAAGT ATACCAGCAT CGCCGATTCC AAGAAGTTCA TCAGCTCCTT
4021 TGACAGGATC ATGTACGTGC CCGAGGAGGA TCTGTTCGAG TTTGCCCTGG ACTATAAGAA
4081 CTTCTCTCGC ACAGACGCCG ATTACATCAA GAAGTGGAAG CTGTACTCCT ACGGCAACCG
4141 GATCAGAATC TTCCGGAATC CTAAGAAGAA CAACGTGTTC GACTGGGAGG AGGTGTGCCT
4201 GACCAGCGCC TATAAGGAGC TGTTCAACAA GTACGGCATC AATTATCAGC AGGGCGATAT
4261 CAGAGCCCTG CTGTGCGAGC AGTCCGACAA GGCCTTCTAC TCTAGCTTTA TGGCCCTGAT
4321 GAGCCTGATG CTGCAGATGC GGAACAGCAT CACAGGCCGC ACCGACGTGG ATTTTCTGAT
4381 CAGCCCTGTG AAGAACTCCG ACGGCATCTT CTACGATAGC CGGAACTATG AGGCCCAGGA
4441 GAATGCCATC CTGCCAAAGA ACGCCGACGC CAATGGCGCC TATAACATCG CCAGAAAGGT
4501 GCTGTGGGCC ATCGGCCAGT TCAAGAAGGC CGAGGACGAG AAGCTGGATA AGGTGAAGAT
4561 CGCCATCTCT AACAAGGAGT GGCTGGAGTA CGCCCAGACC AGCGTGAAGC ACAAAAGGCC
4621 GGCGGCCACG AAAAAGGCCG GCCAGGCAAA AAAGAAAAAG GGATCCTACC CATACGATGT
4681 TCCAGATTAC GCTTATCCCT ACGACGTGCC TGATTATGCA TACCCATATG ATGTCCCCGA
4741 CTATGCCTAA Gaattctgca gatatccagc acagtggcgg ccgctcgagt ctagagggcc
4801 cgtttaaacc cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg
4861 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata
4921 aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt
4981 ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt
5041 gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc
5101 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
5161 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
5221 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
5281 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
5341 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
5401 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
5461 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
5521 gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
5581 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
5641 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
5701 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
5761 catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta
5821 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga
5881 gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt
5941 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
6001 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
6061 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac
6121 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
6181 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
6241 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
6301 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
6361 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
6421 caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag
6481 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
6541 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
6601 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
6661 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
6721 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat
6781 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
6841 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
6901 ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
6961 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
7021 cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc
7081 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
7141 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
7201 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
7261 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
7321 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
7381 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
7441 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
7501 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
7561 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
7621 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
7681 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
7741 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
7801 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
7861 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
7921 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
7981 gatccggcaa acaaaccacc gctggtagcg gtttttttgt ttgcaagcag cagattacgc
8041 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
8101 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
8161 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
8221 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
8281 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
8341 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
8401 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
8461 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
8521 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
8581 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
8641 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
8701 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
8761 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
8821 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
8881 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
8941 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
9001 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
9061 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
9121 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
9181 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtc
//
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