| 質粒名稱: | pLb-Cpf1-pGL3-U6-sgRNA |
|---|---|
| 出品公司: | Addgene |
| 目錄編號: | 107682 |
| 存儲實驗室: | Jia Chen |
|
載體骨架基本信息 |
|
| 載體名稱: | pGL3-Basic |
| 完整質粒大小: | 4897 bp |
| 載體類型: | 哺乳細胞表達 |
| 載體抗性: | 氨芐霉素 |
| 生長溫度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷貝數: | 高拷貝 |
|
插入基因信息 |
|
| 插入基因名稱: | 無 |
| 物種來源: | 光合細菌 Rhodopseudomonas palustris |
|
文獻引用方法 |
|
| 材料和方法部分: | pLb-Cpf1-pGL3-U6-sgRNA was a gift from Jia Chen (Addgene plasmid # 107682 ; //n2t.net/addgene:107682 ; RRID:Addgene_107682) |
| 參考文獻部分: | Base editing with a Cpf1-cytidine deaminase fusion. Li X, Wang Y, Liu Y, Yang B, Wang X, Wei J, Lu Z, Zhang Y, Wu J, Huang X, Yang L, Chen J. Nat Biotechnol. 2018 Mar 19. pii: nbt.4102. doi: 10.1038/nbt.4102. 10.1038/nbt.4102 PubMed 29553573 |
買家導航
pLb-Cpf1-pGL3-U6-sgRNA載體質粒基本信息
pLb-Cpf1-pGL3-U6-sgRNA質粒圖譜載體圖譜和pLb-Cpf1-pGL3-U6-sgRNA載體序列質粒序列多克隆位點信息
pLb-Cpf1-pGL3-U6-sgRNA質粒載體簡介
pLb-Cpf1-pGL3-U6-sgRNA質粒序列載體序列
LOCUS pLb-Cpf1-pGL3-U6-sgRNA 4897 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Expresses LbCpf1 crRNA in mammalian cells.
ACCESSION .
VERSION .
KEYWORDS pLb-Cpf1-pGL3-U6-sgRNA
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4897)
AUTHORS Li X, Wang Y, Liu Y, Yang B, Wang X, Wei J, Lu Z, Zhang Y, Wu J,
Huang X, Yang L, Chen J
TITLE Base editing with a Cpf1-cytidine deaminase fusion.
JOURNAL Nat Biotechnol. 2018 Mar 19. pii: nbt.4102. doi: 10.1038/nbt.4102.
PUBMED 29553573
REFERENCE 2 (bases 1 to 4897)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
//gt12365.cn
FEATURES Location/Qualifiers
source 1..4897
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 61..301
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA"
primer_bind 61..81
/label=hU6-F
/note="Human U6 promoter, forward primer"
primer_bind 232..251
/label=LKO.1 5'
/note="Human U6 promoter, forward primer"
misc_feature 405..522
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 571..1081
/label=hPGK promoter
/note="human phosphoglycerate kinase 1 promoter"
primer_bind 1000..1018
/label=hPGK-F
/note="Human PGK promoter, forward primer"
CDS 1103..1702
/codon_start=1
/gene="pac from Streptomyces alboniger"
/product="puromycin N-acetyltransferase"
/label=PuroR
/note="confers resistance to puromycin"
/translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
primer_bind complement(1103..1122)
/label=Puro-R
/note="Puromycin resistance gene, reverse primer. Also
called puro-variant-R"
primer_bind 1599..1619
/label=Puro-F
/note="Puromycin resistance gene, forward primer"
polyA_signal 1849..1970
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(1860..1879)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 1914..1933
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind complement(2218..2235)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(2389..2977)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(2469..2488)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(3148..4008)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind 3771..3790
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(4009..4113)
/gene="bla"
/label=AmpR promoter
rep_origin 4140..4595
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(4227..4246)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 4437..4458
/label=F1ori-F
/note="F1 origin, forward primer"
polyA_signal 4726..4774
/note="synthetic polyadenylation signal"
misc_feature 4788..4879
/label=pause site
/note="RNA polymerase II transcriptional pause signal from
the human alpha-2 globin gene"
primer_bind 4828..4847
/label=RVprimer3
/note="pGL3 vector, forward primer"
ORIGIN
1 gtgaacggat ctcgacggta tcgatcacga gactagcctc gagcggccgc ccccttcacc
61 gagggcctat ttcccatgat tccttcatat ttgcatatac gatacaaggc tgttagagag
121 ataattggaa ttaatttgac tgtaaacaca aagatattag tacaaaatac gtgacgtaga
181 aagtaataat ttcttgggta gtttgcagtt ttaaaattat gttttaaaat ggactatcat
241 atgcttaccg taacttgaaa gtatttcgat ttcttggctt tatatatctt gtggaaagga
301 cgaaacaccg aatttctact aagtgtagat cgagacccga gagagggtcg gtctcctttt
361 tttgaattct cgacctcgag acaaatggca gtattcatcc acaattttaa aagaaaaggg
421 gggattgggg ggtacagtgc aggggaaaga atagtagaca taatagcaac agacatacaa
481 actaaagaat tacaaaaaca aattacaaaa attcaaaatt ttcgggttta ttacagggac
541 agcagagatc cactttggcc gcggctcgag ggggttgggg ttgcgccttt tccaaggcag
601 ccctgggttt gcgcagggac gcggctgctc tgggcgtggt tccgggaaac gcagcggcgc
661 cgaccctggg actcgcacat tcttcacgtc cgttcgcagc gtcacccgga tcttcgccgc
721 tacccttgtg ggccccccgg cgacgcttcc tgctccgccc ctaagtcggg aaggttcctt
781 gcggttcgcg gcgtgccgga cgtgacaaac ggaagccgca cgtctcacta gtaccctcgc
841 agacggacag cgccagggag caatggcagc gcgccgaccg cgatgggctg tggccaatag
901 cggctgctca gcagggcgcg ccgagagcag cggccgggaa ggggcggtgc gggaggcggg
961 gtgtggggcg gtagtgtggg ccctgttcct gcccgcgcgg tgttccgcat tctgcaagcc
1021 tccggagcgc acgtcggcag tcggctccct cgttgaccga atcaccgacc tctctcccca
1081 gggggatcca ccggagctta ccatgaccga gtacaagccc acggtgcgcc tcgccacccg
1141 cgacgacgtc cccagggccg tacgcaccct cgccgccgcg ttcgccgact accccgccac
1201 gcgccacacc gtcgatccgg accgccacat cgagcgggtc accgagctgc aagaactctt
1261 cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt
1321 ggcggtctgg accacgccgg agagcgtcga agcgggggcg gtgttcgccg agatcggccc
1381 gcgcatggcc gagttgagcg gttcccggct ggccgcgcag caacagatgg aaggcctcct
1441 ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc accgtcggcg tctcgcccga
1501 ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg
1561 cgccggggtg cccgccttcc tggaaacctc cgcgccccgc aacctcccct tctacgagcg
1621 gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat
1681 gacccgcaag cccggtgcct gacgcccgcc ccacgacccg cagcgcccga ccgaaaggag
1741 cgcacgaccc catgcatcgg tacctttaag accaatgact tacaaggcag ctgtagatct
1801 tagccacttt ctagagtcgg ggcggccggc cgcttcgagc agacatgata agatacattg
1861 atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt
1921 gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca
1981 attgcattca ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt
2041 aaaacctcta caaatgtggt aaaatcgata aggatccgtc gaccgatgcc cttgagagcc
2101 ttcaacccag tcagctcctt ccggtgggcg cggggcatga ctatcgtcgc cgcacttatg
2161 actgtcttct ttatcatgca actcgtagga caggtgccgg cagcgctctt ccgcttcctc
2221 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
2281 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
2341 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
2401 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
2461 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
2521 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
2581 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
2641 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
2701 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
2761 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
2821 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
2881 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
2941 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
3001 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
3061 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
3121 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
3181 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
3241 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcggg acccacgctc
3301 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
3361 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
3421 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
3481 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
3541 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
3601 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
3661 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
3721 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
3781 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
3841 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
3901 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
3961 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
4021 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
4081 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
4141 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
4201 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
4261 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
4321 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
4381 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
4441 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
4501 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
4561 cgcgaatttt aacaaaatat taacgcttac aatttgccat tcgccattca ggctgcgcaa
4621 ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagccca agctaccatg
4681 ataagtaagt aatattaagg tacgggaggt acttggagcg gccgcaataa aatatcttta
4741 ttttcattac atctgtgtgt tggttttttg tgtgaatcga tagtactaac atacgctctc
4801 catcaaaaca aaacgaaaca aaacaaacta gcaaaatagg ctgtccccag tgcaagtgca
4861 ggtgccagaa catttctcta tcgataggta ccgatta
//