| 质粒名称: | pAs-Cpf1-pGL3-U6-sgRNA |
|---|---|
| 出品公司： | addgene |
| 目录编号: | 107683 |
| 存储实验室: | Jia Chen |
|
载体骨架基本信息 |
|
| 载体名称: | pGL3-Basic |
| 完整质粒大小: | 4986bp |
| 载体类型: | 哺乳细胞表达 |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | None |
|
文献引用方法 |
|
| 材料和方法部分: | pAs-Cpf1-pGL3-U6-sgRNA was a gift from Jia Chen (Addgene plasmid # 107683 ; http://n2t.net/addgene:107683 ; RRID:Addgene_107683) |
| 参考文献部分: | 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 |
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 pAs-Cpf1-pGL3-U6-sgRNA 4896 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Expresses AsCpf1 crRNA in mammalian cells.
ACCESSION .
VERSION .
KEYWORDS pAs-Cpf1-pGL3-U6-sgRNA
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4896)
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 4896)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..4896
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 69..309
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA"
primer_bind 69..89
/label=hU6-F
/note="Human U6 promoter, forward primer"
primer_bind 240..259
/label=LKO.1 5'
/note="Human U6 promoter, forward primer"
misc_feature 412..529
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 578..1088
/label=hPGK promoter
/note="human phosphoglycerate kinase 1 promoter"
primer_bind 1007..1025
/label=hPGK-F
/note="Human PGK promoter, forward primer"
CDS 1110..1709
/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(1110..1129)
/label=Puro-R
/note="Puromycin resistance gene, reverse primer. Also
called puro-variant-R"
primer_bind 1606..1626
/label=Puro-F
/note="Puromycin resistance gene, forward primer"
polyA_signal 1856..1977
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(1867..1886)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 1921..1940
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind complement(2225..2242)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(2396..2984)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(2476..2495)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(3155..4015)
/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 3778..3797
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(4016..4120)
/gene="bla"
/label=AmpR promoter
rep_origin 4147..4602
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(4234..4253)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 4444..4465
/label=F1ori-F
/note="F1 origin, forward primer"
polyA_signal 4733..4781
/note="synthetic polyadenylation signal"
misc_feature 4795..4886
/label=pause site
/note="RNA polymerase II transcriptional pause signal from
the human alpha-2 globin gene"
primer_bind 4835..4854
/label=RVprimer3
/note="pGL3 vector, forward primer"
ORIGIN
1 accgattagt gaacggatct cgacggtatc gatcacgaga ctagcctcga gcggccgccc
61 ccttcaccga gggcctattt cccatgattc cttcatattt gcatatacga tacaaggctg
121 ttagagagat aattggaatt aatttgactg taaacacaaa gatattagta caaaatacgt
181 gacgtagaaa gtaataattt cttgggtagt ttgcagtttt aaaattatgt tttaaaatgg
241 actatcatat gcttaccgta acttgaaagt atttcgattt cttggcttta tatatcttgt
301 ggaaaggacg aaacaccgaa tttctactct tgtagatcga gacccgagag agggtcggtc
361 tccttttttt gaattctcga cctcgagaca aatggcagta ttcatccaca attttaaaag
421 aaaagggggg attggggggt acagtgcagg ggaaagaata gtagacataa tagcaacaga
481 catacaaact aaagaattac aaaaacaaat tacaaaaatt caaaattttc gggtttatta
541 cagggacagc agagatccac tttggccgcg gctcgagggg gttggggttg cgccttttcc
601 aaggcagccc tgggtttgcg cagggacgcg gctgctctgg gcgtggttcc gggaaacgca
661 gcggcgccga ccctgggact cgcacattct tcacgtccgt tcgcagcgtc acccggatct
721 tcgccgctac ccttgtgggc cccccggcga cgcttcctgc tccgccccta agtcgggaag
781 gttccttgcg gttcgcggcg tgccggacgt gacaaacgga agccgcacgt ctcactagta
841 ccctcgcaga cggacagcgc cagggagcaa tggcagcgcg ccgaccgcga tgggctgtgg
901 ccaatagcgg ctgctcagca gggcgcgccg agagcagcgg ccgggaaggg gcggtgcggg
961 aggcggggtg tggggcggta gtgtgggccc tgttcctgcc cgcgcggtgt tccgcattct
1021 gcaagcctcc ggagcgcacg tcggcagtcg gctccctcgt tgaccgaatc accgacctct
1081 ctccccaggg ggatccaccg gagcttacca tgaccgagta caagcccacg gtgcgcctcg
1141 ccacccgcga cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc gccgactacc
1201 ccgccacgcg ccacaccgtc gatccggacc gccacatcga gcgggtcacc gagctgcaag
1261 aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg
1321 ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga
1381 tcggcccgcg catggccgag ttgagcggtt cccggctggc cgcgcagcaa cagatggaag
1441 gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct
1501 cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg
1561 ccgagcgcgc cggggtgccc gccttcctgg aaacctccgc gccccgcaac ctccccttct
1621 acgagcggct cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct
1681 ggtgcatgac ccgcaagccc ggtgcctgac gcccgcccca cgacccgcag cgcccgaccg
1741 aaaggagcgc acgaccccat gcatcggtac ctttaagacc aatgacttac aaggcagctg
1801 tagatcttag ccactttcta gagtcggggc ggccggccgc ttcgagcaga catgataaga
1861 tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt
1921 gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa acaagttaac
1981 aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga ggttttttaa
2041 agcaagtaaa acctctacaa atgtggtaaa atcgataagg atccgtcgac cgatgccctt
2101 gagagccttc aacccagtca gctccttccg gtgggcgcgg ggcatgacta tcgtcgccgc
2161 acttatgact gtcttcttta tcatgcaact cgtaggacag gtgccggcag cgctcttccg
2221 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
2281 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
2341 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
2401 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
2461 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
2521 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
2581 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
2641 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
2701 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
2761 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
2821 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
2881 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
2941 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
3001 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
3061 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
3121 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
3181 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
3241 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgggacc
3301 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
3361 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
3421 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
3481 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
3541 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
3601 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
3661 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
3721 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
3781 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
3841 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
3901 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
3961 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
4021 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
4081 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
4141 cacctgacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
4201 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
4261 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
4321 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
4381 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
4441 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
4501 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
4561 aatttaacgc gaattttaac aaaatattaa cgcttacaat ttgccattcg ccattcaggc
4621 tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagcccaagc
4681 taccatgata agtaagtaat attaaggtac gggaggtact tggagcggcc gcaataaaat
4741 atctttattt tcattacatc tgtgtgttgg ttttttgtgt gaatcgatag tactaacata
4801 cgctctccat caaaacaaaa cgaaacaaaa caaactagca aaataggctg tccccagtgc
4861 aagtgcaggt gccagaacat ttctctatcg ataggt
//
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