| 质粒名称: | pcg 2.0-BFP-circ-arRNA backbone |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 213965 |
| 存储实验室: | Wensheng Wei |
|
载体骨架基本信息 |
|
| 载体名称: | pcg2.0 |
| 载体出品公司: | Clontech |
| 完整质粒大小: | 7695bp |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | BFP |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 未知 |
|
文献引用方法 |
|
| 材料和方法部分: | pcg 2.0-BFP-circ-arRNA backbone was a gift from Wensheng Wei (Addgene plasmid # 213965 ; http://n2t.net/addgene:213965 ; RRID:Addgene_213965) |
| 参考文献部分: | Engineered circular ADAR-recruiting RNAs increase the efficiency and fidelity of RNA editing in vitro and in vivo. Yi Z, Qu L, Tang H, Liu Z, Liu Y, Tian F, Wang C, Zhang X, Feng Z, Yu Y, Yuan P, Yi Z, Zhao Y, Wei W. Nat Biotechnol. 2022 Jun;40(6):946-955. doi: 10.1038/s41587-021-01180-3. Epub 2022 Feb 10. 10.1038/s41587-021-01180-3 PubMed 35145313 |
买家导航
pcg 2.0-BFP-circ-arRNA backbone载体质粒基本信息
pcg 2.0-BFP-circ-arRNA backbone质粒图谱载体图谱和pcg 2.0-BFP-circ-arRNA backbone载体序列质粒序列多克隆位点信息
pcg 2.0-BFP-circ-arRNA backbone质粒载体简介
pcg 2.0-BFP-circ-arRNA backbone质粒序列载体序列
LOCUS pcg 2.0-BFP-circ-arRNA backbone 7695 bp DNA circular SYN 14-APR-2025
DEFINITION pcg 2.0-BFP-circ-arRNA backbone included a Twister P3 U2A, 5'
ligation sequence, a 3' ligation sequence and Twister P1 based on
pLenti-sgRNA-lib 2.0 backbone (Addgene no. 89638). Then, the
sequences o.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7695)
AUTHORS Yi Z, Qu L, Tang H, Liu Z, Liu Y, Tian F, Wang C, Zhang X, Feng Z,
Yu Y, Yuan P, Yi Z, Zhao Y, Wei W
TITLE Engineered circular ADAR-recruiting RNAs increase the efficiency and
fidelity of RNA editing in vitro and in vivo.
JOURNAL Nat Biotechnol. 2022 Jun;40(6):946-955. doi:
10.1038/s41587-021-01180-3. Epub 2022 Feb 10.
PUBMED 35145313
REFERENCE 2 (bases 1 to 7695)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Apr 14, 2025 from SnapGene Server 8.0.1
https://www.biofeng.com
COMMENT SGRef: number: 1; type: "Journal Article"; doi:
"10.1038/s41587-021-01180-3"; journalName: "Nat Biotechnol"; date:
"2022-06"; volume: "40"; issue: "6"; pages: "946-955"
COMMENT Sequence Label: pcg 2.0-BFP-circ-arRNA backbone
FEATURES Location/Qualifiers
source 1..7695
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind complement(47..66)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 238..617
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 619..817
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
LTR 835..1015
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 1062..1187
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1684..1917
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 2102..2146
/codon_start=1
/product="antigenic peptide corresponding to amino acids
655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik
et al., 2013)"
/label=gp41 peptide
/note="recognized by the 2H10 single-chain llama nanobody"
/translation="KNEQELLELDKWASL"
misc_feature 2444..2561
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 2641..2889
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA
(Domitrovich & Kunkel, 2003)"
primer_bind 2641..2661
/label=hU6-F
/note="Human U6 promoter, forward primer"
primer_bind 2812..2831
/label=LKO.1 5'
/note="Human U6 promoter, forward primer"
primer_bind 3078..3094
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(3136..3169)
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (ATGTATGC) (Shaw et al., 2021)."
enhancer 3285..3588
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 3589..3792
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
CDS 3827..4528
/codon_start=1
/product="monomeric blue fluorescent protein"
/label=TagBFP
/note="mammalian codon-optimized"
/translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
DLPSKLGHKLN"
misc_feature 4584..5172
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(4637..4657)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(5055..5066)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
primer_bind complement(5175..5191)
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
LTR 5701..5881
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
rep_origin complement(5943..6531)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(6023..6042)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(join(6702..7493,7494..7562))
/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 7325..7344
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(7563..7667)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
61 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
121 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
181 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac
241 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
301 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
361 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
421 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
481 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
541 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
601 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
661 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
721 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
781 gcggtaggcg tgtacggtgg gaggtctata taagcagcgc gttttgcctg tactgggtct
841 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
901 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
961 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc
1021 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc
1081 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa
1141 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg
1201 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata
1261 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc
1321 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga
1381 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc
1441 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca
1501 aaagtaagac caccgcacag caagcggccg gccgctgatc ttcagacctg gaggaggaga
1561 tatgagggac aattggagaa gtgaattata taaatataaa gtagtaaaaa ttgaaccatt
1621 aggagtagca cccaccaagg caaagagaag agtggtgcag agagaaaaaa gagcagtggg
1681 aataggagct ttgttccttg ggttcttggg agcagcagga agcactatgg gcgcagcgtc
1741 aatgacgctg acggtacagg ccagacaatt attgtctggt atagtgcagc agcagaacaa
1801 tttgctgagg gctattgagg cgcaacagca tctgttgcaa ctcacagtct ggggcatcaa
1861 gcagctccag gcaagaatcc tggctgtgga aagataccta aaggatcaac agctcctggg
1921 gatttggggt tgctctggaa aactcatttg caccactgct gtgccttgga atgctagttg
1981 gagtaataaa tctctggaac agatttggaa tcacacgacc tggatggagt gggacagaga
2041 aattaacaat tacacaagct taatacactc cttaattgaa gaatcgcaaa accagcaaga
2101 aaagaatgaa caagaattat tggaattaga taaatgggca agtttgtgga attggtttaa
2161 cataacaaat tggctgtggt atataaaatt attcataatg atagtaggag gcttggtagg
2221 tttaagaata gtttttgctg tactttctat agtgaataga gttaggcagg gatattcacc
2281 attatcgttt cagacccacc tcccaacccc gaggggaccc gacaggcccg aaggaataga
2341 agaagaaggt ggagagagag acagagacag atccattcga ttagtgaacg gatcggcact
2401 gcgtgcgcca attctgcaga caaatggcag tattcatcca caattttaaa agaaaagggg
2461 ggattggggg gtacagtgca ggggaaagaa tagtagacat aatagcaaca gacatacaaa
2521 ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt tcgggtttat tacagggaca
2581 gcagagatcc agtttggtta gtaccgggcc cgctctagag gtaccaaggt ctggcaggaa
2641 gagggcctat ttcccatgat tccttcatat ttgcatatac gatacaaggc tgttagagag
2701 ataattagaa ttaatttgac tgtaaacaca aagatattag tacaaaatac gtgacgtaga
2761 aagtaataat ttcttgggta gtttgcagtt ttaaaattat gttttaaaat ggactatcat
2821 atgcttaccg taacttgaaa gtatttcgat ttcttggctt tatatatctt gtggaaagga
2881 cgaaacaccg gccatcagtc gccggtccca agcccggata aaatgggagg gggcgggaaa
2941 ccgcctaacc atgccgactg atggcagaga gacgatatta cgtctcactg ccatcagtcg
3001 gcgtggactg tagaacactg ccaatgccgg tcccaagccc ggataaaagt ggagggtaca
3061 gtccacgctt tttttctcga ggtcgacggt atcgataagc tcgcttcacg agattccagc
3121 aggtcgaggg acctaataac ttcgtatagc atacattata cgaagttata ttaagggttc
3181 caagcttaag cggccgcgtg gataaccgta ttaccgccat gcattagtta ttaatagtaa
3241 tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg
3301 gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg
3361 tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta
3421 cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt
3481 gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac
3541 tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt
3601 tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac
3661 cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt
3721 cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat
3781 ataagcagag ctggtttagt gaaccgtcag atccgctagc gccaccatga gcgagctgat
3841 taaggagaac atgcacatga agctgtacat ggagggcacc gtggacaacc atcacttcaa
3901 gtgcacatcc gagggcgaag gcaagcccta cgagggcacc cagaccatga gaatcaaggt
3961 ggtcgagggc ggccctctcc ccttcgcctt cgacatcctg gctactagct tcctctacgg
4021 cagcaagacc ttcatcaacc acacccaggg catccccgac ttcttcaagc agtccttccc
4081 tgagggcttc acatgggaga gagtcaccac atacgaagac gggggcgtgc tgaccgctac
4141 ccaggacacc agcctccagg acggctgcct catctacaac gtcaagatca gaggggtgaa
4201 cttcacatcc aacggccctg tgatgcagaa gaaaacactc ggctgggagg ccttcaccga
4261 gactctgtac cccgctgacg gcggcctgga aggcagaaac gacatggccc tgaagctcgt
4321 gggcgggagc catctgatcg caaacatcaa gaccacatat agatccaaga aacccgctaa
4381 gaacctcaag atgcctggcg tctactatgt ggactacaga ctggaaagaa tcaaggaggc
4441 caacaacgag acctacgtcg agcagcacga ggtggcagtg gccagatact gcgacctccc
4501 tagcaaactg gggcacaaac tcaattaaac atgtttaagg gttccggttc cactaggtac
4561 aattcgatat caagcttatc gataatcaac ctctggatta caaaatttgt gaaagattga
4621 ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt
4681 tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt
4741 tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg
4801 tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg
4861 ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc
4921 gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaaat
4981 catcgtcctt tccttggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct
5041 tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg
5101 ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg
5161 ccgcctcccc gcatcgatac cgtcgacctc gatcgagacc tagaaaaaca tggagcaatc
5221 acaagtagca atacagcagc taccaatgct gattgtgcct ggctagaagc acaagaggag
5281 gaggaggtgg gttttccagt cacacctcag gtacctttaa gaccaatgac ttacaaggca
5341 gctgtagatc ttagccactt tttaaaagaa aaggggggac tggaagggct aattcactcc
5401 caacgaagac aagatatcct tgatctgtgg atctaccaca cacaaggcta cttccctgat
5461 tggcagaact acacaccagg gccagggatc agatatccac tgacctttgg atggtgctac
5521 aagctagtac cagttgagca agagaaggta gaagaagcca atgaaggaga gaacacccgc
5581 ttgttacacc ctgtgagcct gcatgggatg gatgacccgg agagagaagt attagagtgg
5641 aggtttgaca gccgcctagc atttcatcac atggcccgag agctgcatcc ggactgtact
5701 gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca
5761 ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg
5821 tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc
5881 agcatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
5941 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
6001 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
6061 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
6121 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
6181 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
6241 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
6301 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
6361 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
6421 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
6481 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
6541 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
6601 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
6661 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
6721 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
6781 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
6841 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
6901 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
6961 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
7021 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
7081 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
7141 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
7201 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
7261 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
7321 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
7381 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
7441 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
7501 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
7561 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
7621 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
7681 aaagtgccac ctgac
//
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