| 质粒名称: | prCO |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 153960 |
| 存储实验室: | Hiroyuki Konishi |
|
载体骨架基本信息 |
|
| 载体名称: | pBABEneo |
| 载体出品公司: | Clontech |
| 空载体大小: | 5330bp |
| 完整质粒大小: | 7308bp |
| 载体类型: | 哺乳细胞表达, Retroviral ; A reporter plasmid detecting DNA recombination |
| 筛选标记: | 新霉素（G418） ; NeoR/KanR |
| 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: | 5' and 3' EGFP fragments sharing a 386-bp sequence, separated by a NeoR gene cassette and arranged in the same direction |
| 物种来源: | Synthetic |
| 插入大小: | 3144bp |
| 基因突变: | No mutation |
| 启动子: | LTR |
| 标签/融合蛋白: | NLS-ZF (N terminal on insert) |
| 克隆方法: | 未知 |
|
文献引用方法 |
|
| 材料和方法部分: | prCO was a gift from Hiroyuki Konishi (Addgene plasmid # 153960 ; http://n2t.net/addgene:153960 ; RRID:Addgene_153960) |
| 参考文献部分: | Tandem Paired Nicking Promotes Precise Genome Editing with Scarce Interference by p53. Hyodo T, Rahman ML, Karnan S, Ito T, Toyoda A, Ota A, Wahiduzzaman M, Tsuzuki S, Okada Y, Hosokawa Y, Konishi H. Cell Rep. 2020 Jan 28;30(4):1195-1207.e7. doi: 10.1016/j.celrep.2019.12.064. 10.1016/j.celrep.2019.12.064 PubMed 31995758 |
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 prCO 7224 bp DNA circular SYN 26-AUG-2024
DEFINITION Used to detect crossover-type DNA recombination events elicited by
iso-positional nicking of two homologous DNA fragments
(recombination analogous to Holliday's model).
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7224)
AUTHORS Hyodo T, Rahman ML, Karnan S, Ito T, Toyoda A, Ota A, Wahiduzzaman
M, Tsuzuki S, Okada Y, Hosokawa Y, Konishi H
TITLE Tandem Paired Nicking Promotes Precise Genome Editing with Scarce
Interference by p53.
JOURNAL Cell Rep. 2020 Jan 28;30(4):1195-1207.e7. doi:
10.1016/j.celrep.2019.12.064.
PUBMED 31995758
REFERENCE 2 (bases 1 to 7224)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3
https://www.biofeng.com
COMMENT SGRef: number: 1; type: "Journal Article"; doi:
"10.1016/j.celrep.2019.12"; journalName: "Cell Rep"; date:
"2020-01-28- 28"; volume: "30"; issue: "4"; pages: "1195-1207.e7"
FEATURES Location/Qualifiers
source 1..7224
/mol_type="other DNA"
/organism="synthetic DNA construct"
LTR 8..477
/note="long terminal repeat from Moloney murine leukemia
virus"
misc_feature 542..741
/label=MMLV Psi
/note="packaging signal of Moloney murine leukemia virus
(MMLV)"
CDS 942..1358
/label=gag (truncated)
/note="truncated Moloney murine leukemia virus (MMLV) gag
gene lacking the start codon"
primer_bind 1288..1310
/label=pLXSN 5'
/note="Murine stem cell virus, forward primer. Also called
MSCV"
primer_bind 1326..1342
/label=pBABE 5'
/note="Psi packaging signal, forward primer for pBABE
vectors"
CDS 1423..1443
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 1465..1485
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
regulatory 1699..1708
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS 1705..2169
/codon_start=1
/product="improved N-terminal fragment of mVenus for use in
bimolecular fluorescence complementation (BiFC) (Kodama and
Hu, 2010)"
/label=VN155(I152L)
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMA"
primer_bind complement(1750..1771)
/label=EGFP-N
/note="EGFP, reverse primer"
intron 2278..2507
/label=chimeric intron
/note="chimera between introns from adenovirus and
immunoglobulin heavy chain genes"
primer_bind 2490..2509
/label=pMT2-F
/note="Synthetic intron, forward primer"
misc_feature 2636..3098
/label=IRES
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
primer_bind complement(2692..2709)
/label=IRES reverse
/note="IRES internal ribosome entry site, reverse primer.
Also called pCDH-rev"
primer_bind 2919..2938
/label=IRES-F
/note="IRES internal ribosome entry site, forward primer"
CDS 3112..3906
/codon_start=1
/gene="aph(3')-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/label=NeoR/KanR
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MAEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKARMPDGDDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(3166..3185)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 3776..3795
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind 4484..4505
/label=EGFP-C
/note="EGFP, forward primer"
LTR 4718..5187
/note="long terminal repeat from Moloney murine leukemia
virus"
primer_bind complement(5317..5334)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(5488..6076)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(5568..5587)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(6236..7096)
/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 6859..6878
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(7097..7201)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 ctgcagcctg aatatgggcc aaacaggata tctgtggtaa gcagttcctg ccccggctca
61 gggccaagaa cagatggaac agctgaatat gggccaaaca ggatatctgt ggtaagcagt
121 tcctgccccg gctcagggcc aagaacagat ggtccccaga tgcggtccag ccctcagcag
181 tttctagaga accatcagat gtttccaggg tgccccaagg acctgaaatg accctgtgcc
241 ttatttgaac taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga
301 gctcaataaa agagcccaca acccctcact cggggcgcca gtcctccgat tgactgagtc
361 gcccgggtac ccgtgtatcc aataaaccct cttgcagttg catccgactt gtggtctcgc
421 tgttccttgg gagggtctcc tctgagtgat tgactacccg tcagcggggg tctttcattt
481 gggggctcgt ccgggatcgg gagacccctg cccagggacc accgacccac caccgggagg
541 caagctggcc agcaacttat ctgtgtctgt ccgattgtct agtgtctatg actgatttta
601 tgcgcctgcg tcggtactag ttagctaact agctctgtat ctggcggacc cgtggtggaa
661 ctgacgagtt ctgaacaccc ggccgcaacc ctgggagacg tcccagggac tttgggggcc
721 gtttttgtgg cccgacctga ggaagggagt cgatgtggaa tccgaccccg tcaggatatg
781 tggttctggt aggagacgag aacctaaaac agttcccgcc tccgtctgaa tttttgcttt
841 cggtttggaa ccgaagccgc gcgtcttgtc tgctgcagcg ctgcagcatc gttctgtgtt
901 gtctctgtct gactgtgttt ctgtatttgt ctgaaaatta gggccagact gttaccactc
961 ccttaagttt gaccttaggt cactggaaag atgtcgagcg gatcgctcac aaccagtcgg
1021 tagatgtcaa gaagagacgt tgggttacct tctgctctgc agaatggcca acctttaacg
1081 tcggatggcc gcgagacggc acctttaacc gagacctcat cacccaggtt aagatcaagg
1141 tcttttcacc tggcccgcat ggacacccag accaggtccc ctacatcgtg acctgggaag
1201 ccttggcttt tgacccccct ccctgggtca agccctttgt acaccctaag cctccgcctc
1261 ctcttcctcc atccgccccg tctctccccc ttgaacctcc tcgttcgacc ccgcctcgat
1321 cctcccttta tccagccctc actccttctc taggcgccgg ccggatccca gtgtggtggt
1381 acgtaggaat tcagatccgc cgccactatg ggatcaagat cgccaaaaaa gaagagaaag
1441 gtgccgaaga agcatgcagc accaccaaaa aaaaaacgaa aagtagaaga cccacgaggc
1501 aacaccagcg gcgtgctgag cacccccaag gccaagaggg ccaagcaccc ccccggcacc
1561 gagaagccca ggagcaggag ccagagcgag cagcccgcca cctgccccat ctgctacgcc
1621 gtgatcaggc agagcaggaa cctgaggagg cacctggagc tgaggcactt cgccaagccc
1681 ggcgtggatc caccggtcgc caccatggtg agcaagggcg aggagctgtt caccggggtg
1741 gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc
1801 gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc
1861 aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc
1921 agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc
1981 tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag
2041 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag
2101 gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat
2161 atcatggccg acaagcagaa gaacggcatc aggtgaactt caataaaagc tggccgcata
2221 gataactgat ccagtgtgct ggaattaatt cgctgtctgc gagggccagc tgttggggtg
2281 agtactccct ctcaaaagcg ggcatgactt ctgcgctaag attgtcagtt tccaaaaacg
2341 aggaggattt gatattcacc tggcccgcgg tgatgccttt gagggtggcc gcgtccatct
2401 ggtcagaaaa gacaatcttt ttgttgtcaa gcttgaggtg tggcaggctt gagatctggc
2461 catacacttg agtgacaatg acatccactt tgcctttctc tccacaggtg tccactccca
2521 ggtccaactg caggtcgagc gtcgagaacg ttactggccg aagccgcttg gaataaggcc
2581 ggtgtgcgtt tgtctatatg ttattttcca ccatattgcc gtcttttggc aatgtgaggg
2641 cccggaaacc tggccctgtc ttcttgacga gcattcctag gggtctttcc cctctcgcca
2701 aaggaatgca aggtctgttg aatgtcgtga aggaagcagt tcctctggaa gcttcttgaa
2761 gacaaacaac gtctgtagcg accctttgca ggcagcggaa ccccccacct ggcgacaggt
2821 gcctctgcgg ccaaaagcca cgtgtataag atacacctgc aaaggcggca caaccccagt
2881 gccacgttgt gagttggata gttgtggaaa gagtcaaatg gctctcctca agcgtattca
2941 acaaggggct gaaggatgcc cagaaggtac cccattgtat gggatctgat ctggggcctc
3001 ggtgcacatg ctttacatgt gtttagtcga ggttaaaaaa acgtctaggc cccccgaacc
3061 acggggacgt ggttttcctt tgaaaaacac gatgataagc ttgccacaac catggctgaa
3121 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
3181 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
3241 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag
3301 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
3361 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
3421 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
3481 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
3541 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
3601 gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgatgat
3661 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
3721 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
3781 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
3841 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
3901 ttctgagggg atcaattctc tagagctagg gataacaggg taatacctac ggcaagctga
3961 ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca
4021 ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact
4081 tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg
4141 acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca
4201 tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt
4261 acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg
4321 tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc
4381 agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca
4441 cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt
4501 tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtaa atcgataaaa
4561 taaaagattt tatttagtct ccagaaaaag gggggaatga aagaccccac ctgtaggttt
4621 ggcaagctag cttaagtaac gccattttgc aaggcatgga aaaatacata actgagaata
4681 gagaagttca gatcaaggtc aggaacagat ggaacagctg aatatgggcc aaacaggata
4741 tctgtggtaa gcagttcctg ccccggctca gggccaagaa cagatggaac agctgaatat
4801 gggccaaaca ggatatctgt ggtaagcagt tcctgccccg gctcagggcc aagaacagat
4861 ggtccccaga tgcggtccag ccctcagcag tttctagaga accatcagat gtttccaggg
4921 tgccccaagg acctgaaatg accctgtgcc ttatttgaac taaccaatca gttcgcttct
4981 cgcttctgtt cgcgcgcttc tgctccccga gctcaataaa agagcccaca acccctcact
5041 cggggcgcca gtcctccgat tgactgagtc gcccgggtac ccgtgtatcc aataaaccct
5101 cttgcagttg catccgactt gtggtctcgc tgttccttgg gagggtctcc tctgagtgat
5161 tgactacccg tcagcggggg tctttcacat gcagcatgta tcaaaattaa tttggttttt
5221 tttcttaagt atttacatta aatggccata gttgcattaa tgaatcggcc aacgcgcggg
5281 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
5341 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
5401 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
5461 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
5521 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
5581 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
5641 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
5701 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
5761 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
5821 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
5881 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
5941 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
6001 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
6061 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
6121 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
6181 ccttttgcgg ccgcaaatca atctaaagta tatatgagta aacttggtct gacagttacc
6241 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
6301 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
6361 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
6421 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
6481 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
6541 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
6601 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
6661 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
6721 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
6781 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
6841 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
6901 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
6961 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
7021 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
7081 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
7141 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
7201 gcacatttcc ccgaaaagtg ccac
//
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