| 质粒名称: | pTubb3-MC |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 87112 |
| 存储实验室: | Juan Belmonte |
|
载体骨架基本信息 |
|
| 载体名称: | pMC.BESPX |
| 载体出品公司: | -- |
| 载体抗性: | 卡那霉素 |
| 空载体大小: | -- |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: |
CRISPR
|
| 筛选标记: | -- |
| 高拷贝/低拷贝: | 高拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | GFP |
| 物种来源: | Synthetic |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | -- |
| 克隆方法: | Ligation Independent 克隆 |
| 5' 克隆位点: | -- |
| 3' 克隆位点: | -- |
| 5' 测序引物 | -- |
| 3' 测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pTubb3-MC was a gift from Juan Belmonte (Addgene plasmid # 87112 ; http://n2t.net/addgene:87112 ; RRID:Addgene_87112) |
| 参考文献部分: | In vivo genome editing via CRISPR/Cas9 mediated homology-independent targeted integration. Suzuki K, Tsunekawa Y, Hernandez-Benitez R, Wu J, Zhu J, Kim EJ, Hatanaka F, Yamamoto M, Araoka T, Li Z, Kurita M, Hishida T, Li M, Aizawa E, Guo S, Chen S, Goebl A, Soligalla RD, Qu J, Jiang T, Fu X, Jafari M, Esteban CR, Berggren WT, Lajara J, Nunez-Delicado E, Guillen P, Campistol JM, Matsuzaki F, Liu GH, Magistretti P, Zhang K, Callaway EM, Zhang K, Belmonte JC. Nature. 2016 Dec 1;540(7631):144-149. doi: 10.1038/nature20565. Epub 2016 Nov 16. 10.1038/nature20565 PubMed 27851729 |
买家导航
pTubb3MC载体基本信息
pTubb3MC载体图谱质粒图谱和多克隆位点信息
pTubb3MC载体简介
pTubb3MC载体序列
LOCUS pTubb3-MC 4885 bp ds-DNA circular SYN 25-JUL-2019
DEFINITION minicircle parental backbone plasmid including GFP knock-in donor
targeting Tubb3 gene and one cutting site for HITI .
ACCESSION .
VERSION .
KEYWORDS pTubb3-MC
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4885)
AUTHORS Suzuki K, Tsunekawa Y, Hernandez-Benitez R, Wu J, Zhu J, Kim EJ,
Hatanaka F, Yamamoto M, Araoka T, Li Z, Kurita M, Hishida T, Li M,
Aizawa E, Guo S, Chen S, Goebl A, Soligalla RD, Qu J, Jiang T, Fu X,
Jafari M, Esteban CR, Berggren WT, Lajara J, Nunez-Delicado E,
Guillen P, Campistol JM, Matsuzaki F, Liu GH, Magistretti P, Zhang
K, Callaway EM, Zhang K, Belmonte JC
TITLE In vivo genome editing via CRISPR/Cas9 mediated homology-independent
targeted integration.
JOURNAL Nature. 2016 Dec 1;540(7631):144-149. doi: 10.1038/nature20565. Epub
2016 Nov 16.
PUBMED 27851729
REFERENCE 2 (bases 1 to 4885)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Jul 25, 2019 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..4885
/organism="synthetic DNA construct"
/mol_type="other DNA"
CDS complement(743..760)
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
protein_bind 781..814
/label=attB
/bound_moiety="phage phi-C31 integrase"
/note="minimal attB site for the phi-C31 integrase (Groth
et al., 2000)"
CDS 862..1575
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/note="mammalian codon-optimized"
/translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
FVTAAGITLGMDELYK"
primer_bind complement(904..925)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(1165..1184)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 1512..1533
/label=EGFP-C
/note="EGFP, forward primer"
CDS complement(1708..2502)
/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="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(1819..1838)
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind 2429..2448
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind complement(2866..2883)
/label=pBAD Reverse
/note="For vectors with E. coli araBAD promoter, reverse
primer"
terminator 3036..3122
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 3214..3241
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
rep_origin 3375..3963
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 3864..3883
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 4117..4134
/label=L4440
/note="L4440 vector, forward primer"
misc_feature 4149..4289
/label=bom
/note="basis of mobility region from pBR322"
primer_bind complement(4215..4234)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 4375..4397
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
ORIGIN
1 acattaccct gttatcccta gatacattac cctgttatcc cagatgacat accctgttat
61 ccctagatga cattaccctg ttatcccaga tgacattacc ctgttatccc tagatacatt
121 accctgttat cccagatgac ataccctgtt atccctagat gacattaccc tgttatccca
181 gatgacatta ccctgttatc cctagataca ttaccctgtt atcccagatg acataccctg
241 ttatccctag atgacattac cctgttatcc cagatgacat taccctgtta tccctagata
301 cattaccctg ttatcccaga tgacataccc tgttatccct agatgacatt accctgttat
361 cccagatgac attaccctgt tatccctaga tacattaccc tgttatccca gatgacatac
421 cctgttatcc ctagatgaca ttaccctgtt atcccagatg acattaccct gttatcccta
481 gatacattac cctgttatcc cagatgacat accctgttat ccctagatga cattaccctg
541 ttatcccaga tgacattacc ctgttatccc tagatacatt accctgttat cccagatgac
601 ataccctgtt atccctagat gacattaccc tgttatccca gatgacatta ccctgttatc
661 cctagataca ttaccctgtt atcccagatg acataccctg ttatccctag atgacattac
721 cctgttatcc cagataaact caatgatgat gatgatgatg gtcgagactc agcggccgcg
781 gtgccagggc gtgcccttgg gctccccggg cgcgactata agctgcgagc aacttcactt
841 gggtatgccg gcggtagcgc tgtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc
901 atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc
961 gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg
1021 cccgtgccct ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc
1081 taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc
1141 caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag
1201 ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac
1261 ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg
1321 gccgacaagc agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac
1381 ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg
1441 ctgctgcccg acaaccacta cctgagcacc cagtccgccc tgagcaaaga ccccaacgag
1501 aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg
1561 gacgagctgt acaaggggcc cgccccaact ggggtaacct ttgagttctc tcagttgggg
1621 gtaatcagca tcatgatgtg gtaccacatc atgatgctga ttataagaat gcggccgcca
1681 cactctagtg gatctcgagt taataattca gaagaactcg tcaagaaggc gatagaaggc
1741 gatgcgctgc gaatcgggag cggcgatacc gtaaagcacg aggaagcggt cagcccattc
1801 gccgccaagc tcttcagcaa tatcacgggt agccaacgct atgtcctgat agcggtccgc
1861 cacacccagc cggccacagt cgatgaatcc agaaaagcgg ccattttcca ccatgatatt
1921 cggcaagcag gcatcgccat gggtcacgac gagatcctcg ccgtcgggca tgctcgcctt
1981 gagcctggcg aacagttcgg ctggcgcgag cccctgatgc tcttcgtcca gatcatcctg
2041 atcgacaaga ccggcttcca tccgagtacg tgctcgctcg atgcgatgtt tcgcttggtg
2101 gtcgaatggg caggtagccg gatcaagcgt atgcagccgc cgcattgcat cagccatgat
2161 ggatactttc tcggcaggag caaggtgaga tgacaggaga tcctgccccg gcacttcgcc
2221 caatagcagc cagtcccttc ccgcttcagt gacaacgtcg agcacagctg cgcaaggaac
2281 gcccgtcgtg gccagccacg atagccgcgc tgcctcgtct tgcagttcat tcagggcacc
2341 ggacaggtcg gtcttgacaa aaagaaccgg gcgcccctgc gctgacagcc ggaacacggc
2401 ggcatcagag cagccgattg tctgttgtgc ccagtcatag ccgaatagcc tctccaccca
2461 agcggccgga gaacctgcgt gcaatccatc ttgttcaatc atgcgaaacg atcctcatcc
2521 tgtctcttga tcagagcttg atcccctgcg ccatcagatc cttggcggca agaaagccat
2581 ccagtttact ttgcagggct tcccaacctt accagagggc gccccagctg gcaattccgg
2641 ttcgcttgct gtccataaaa ccgcccagtc tagctatcgc catgtaagcc cactgcaagc
2701 tacctgcttt ctctttgcgc ttgcgttttc ccttgtccag atagcccagt agctgacatt
2761 catccggggt cagcaccgtt tctgcggact ggctttctac gcgctcgagg gggggccaaa
2821 cggtctccag cttggctgtt ttggcggatg agagaagatt ttcagcctga tacagattaa
2881 atcagaacgc agaagcggtc tgataaaaca gaatttgcct ggcggcagta gcgcggtggt
2941 cccacctgac cccatgccga actcagaagt gaaacgccgt agcgccgatg gtagtgtggg
3001 gtctccccat gcgagagtag ggaactgcca ggcatcaaat aaaacgaaag gctcagtcga
3061 aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctcctg agtaggacaa
3121 atccgccggg agcggatttg aacgttgcga agcaacggcc cggagggtgg cgggcaggac
3181 gcccgccata aactgccagg catcaaatta agcagaaggc catcctgacg gatggccttt
3241 ttgcgtttct acaaactctt ttgtttattt ttctaaatac attcaaatat gtatccgctc
3301 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
3361 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
3421 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
3481 aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag
3541 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
3601 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
3661 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
3721 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
3781 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
3841 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
3901 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
3961 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
4021 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
4081 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
4141 gaagagcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata
4201 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta tacactccgc
4261 tatcgctacg tgactgggtc atggctgcgc cccgacaccc gccaacaccc gctgacgcgc
4321 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga
4381 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag cagatcaatt
4441 cgcgcgcgaa ggcgaagcgg catgcataat gtgcctgtca aatggacgaa gcagggattc
4501 tgcaaaccct atgctactcc gtcaagccgt caattgtctg attcgttacc aattatgaca
4561 acttgacggc tacatcattc actttttctt cacaaccggc acggaactcg ctcgggctgg
4621 ccccggtgca ttttttaaat acccgcgaga aatagagttg atcgtcaaaa ccaacattgc
4681 gaccgacggt ggcgataggc atccgggtgg tgctcaaaag cagcttcgcc tggctgatac
4741 gttggtcctc gcgccagctt aagacgctaa tccctaactg ctggcggaaa agatgtgaca
4801 gacgcgacgg cgacaagcaa acatgctgtg cgacgctggc gattcttctc tcatccgcca
4861 aaacagccaa gctggagacc gtttg
//
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