| 质粒名称: | pAAV-rMERTK-HITI |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 87119 |
| 存储实验室: | Juan Belmonte |
|
载体骨架基本信息 |
|
| 载体名称: | PX551 |
| 载体出品公司: | Addgene 60957 |
| 完整质粒大小: | 7122bp |
| 载体类型: | AAV, CRISPR |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 30℃ |
| 宿主菌: | NEB Stable |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: |
U6-rMertksgRNA-Mertk(intron1-2)-nEF-GFPKASH-pA
gRNA/shRNA sequenceGAGGACCACTGCAACGGGGC
|
| 物种来源: | Synthetic |
| 克隆方法: | Ligation Independent Cloning |
| 5’测序引物 | 未知 |
|
文献引用方法 |
|
| 材料和方法部分: | pAAV-rMERTK-HITI was a gift from Juan Belmonte (Addgene plasmid # 87119 ; http://n2t.net/addgene:87119 ; RRID:Addgene_87119) |
| 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
pAAV-rMERTK-HITI载体质粒基本信息
pAAV-rMERTK-HITI质粒图谱载体图谱和pAAV-rMERTK-HITI载体序列质粒序列多克隆位点信息
pAAV-rMERTK-HITI质粒载体简介
pAAV-rMERTK-HITI质粒序列载体序列
LOCUS pAAV-rMERTK-HITI 7122 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Gene correction donor AAV for RCS rat. AAV backbone plasmid
including exon 2 of rat Mertk and rMertkgRNA for HITI.
ACCESSION .
VERSION .
KEYWORDS pAAV-rMERTK-HITI
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7122)
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 7122)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..7122
/organism="synthetic DNA construct"
/mol_type="other DNA"
repeat_region 1..130
/label=AAV2 ITR (alternate)
/note="Functional equivalent of wild-type AAV2 ITR"
promoter 160..400
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA"
primer_bind 160..180
/label=hU6-F
/note="Human U6 promoter, forward primer"
primer_bind 331..350
/label=LKO.1 5'
/note="Human U6 promoter, forward primer"
misc_RNA 429..504
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
protein_bind complement(516..540)
/gene="mutant version of attB"
/label=attB2
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
promoter 2802..3013
/label=EF-1-alpha core promoter
/note="core promoter for human elongation factor
EF-1-alpha"
LTR 3026..3294
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from human
T-cell leukemia virus (HTLV) type 1"
regulatory 3318..3327
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 3324..4040
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
CDS 3324..4040
/codon_start=1
/product="the original enhanced GFP (Yang et al., 1996)"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(3369..3390)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(3630..3649)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 3977..3998
/label=EGFP-C
/note="EGFP, forward primer"
polyA_signal 4314..4362
/note="synthetic polyadenylation signal"
repeat_region 4385..4525
/label=AAV2 ITR
/note="inverted terminal repeat of adeno-associated virus
serotype 2"
repeat_region 4385..4514
/label=AAV2 ITR
rep_origin 4600..5055
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(4687..4706)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 4897..4918
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(5072..5091)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 5191..5213
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(5251..5269)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 5337..5441
/gene="bla"
/label=AmpR promoter
CDS 5442..6302
/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 complement(5660..5679)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 6473..7061
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 6962..6981
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
121 aggggttcct gcggccgcac gcgtaaggtc gggcaggaag agggcctatt tcccatgatt
181 ccttcatatt tgcatatacg atacaaggct gttagagaga taattagaat taatttgact
241 gtaaacacaa agatattagt acaaaatacg tgacgtagaa agtaataatt tcttgggtag
301 tttgcagttt taaaattatg ttttaaaatg gactatcata tgcttaccgt aacttgaaag
361 tatttcgatt tcttggcttt atatatcttg tggaaaggac gaaacaccga ggaccactgc
421 aacggggcgt tttagagcta gaaatagcaa gttaaaataa ggctagtccg ttatcaactt
481 gaaaaagtgg caccgagtcg gtgctttttt tctagaccca gctttcttgt acaaagttgg
541 cgtttaaacc cagccccgtt gcagtggtcc tcctgccttg ctcaccgcct caaccattca
601 ctgccaacat atttgaaaga acagcaacca gcagagaatg tgtgttccca agtaccgctt
661 acacaagctc aagttcagtg tatatgagtc tctattttta gaaatacttg cagaccagat
721 aaaataaaaa catttcctgc ctcgttcatg tagacatgca tttgctagaa ggtccagatc
781 tggagcctaa gagaactttc cctgttggac acaaaacttc taaacccatg tctttgtgac
841 ccacagtggc cattctggtt gtctgactga acagatgggg agtaggaaag gtgaccctag
901 accctcatat agctgtcact ttcctgctag gttatgtata tagaacttgg agtgagttac
961 cacttgacaa aaattgtcag agaaacttcg ttgtttaaaa tttatatact tggtgtgtat
1021 ataggaggtg gtgcacatcc ttctgtacac atgcggaggt cagaggacac cttgcaggag
1081 tcagttctct cctcccactg tgtgggctct ggagatggag ctcagggtca tcaggctggg
1141 cagtaagcac ctcttaccca ctgagccatc tcaccagccc aggaaattaa cttccttcct
1201 tccttccttc cttccttcct tccttccttc cttccttcct tctttcttcg tttttttttt
1261 ttcccagaga gggtttttct atgtaataga gccctgtcta tcctgtactc gctttgtaga
1321 tcaggctggc cttgaactca cagagatcca actgcctctg tctcctgagt attgggattg
1381 aaggcaagca ccaccacacc aggccagaat tttcttaagt attctttaga acagtggttc
1441 gaaaccttct taatgctgtg accatttggt acagttcctc atattgtgat gactcccaac
1501 cataaaatgt tttcattgct acttcataac tgtggttatt ctgctactgt tatgaatcat
1561 aatgtaaata tttttggaaa tagagctgcc aacgtggagg agacctacag gttgagaagc
1621 acccgggtac acaaattgtt aactgaggat agtcgtcaga tgtgcgtatc tgtcaggaag
1681 ttctccccgc ctcttttctc tttgttaagg tgctaggaac tgaactgggg gactcgtgtg
1741 ggtgaggcag tgttctacca ctgagctaca tcccagctct tgattctttt gggacagatt
1801 ttcactgtgt aactcaggtt ggtcttgagc tctcggccct cctgcctcag cctcacagat
1861 gcttggatga caagctacgc ctttgcatcc aacttcgtgc ttatcttctc tgaggccacg
1921 gaggatgttc tgcttcaccg actggtgtgg tgtcagcatc tgaagttttt ctccatcatc
1981 ttaaaggcta aaactctact attttgcttt gcaggcactg ctgaaaagga ggaagaaatc
2041 aagccagatc agccattttc agggccttta ccagggagcc taccagctga ccacaggcca
2101 ttcttcgccc ctcactccag tggggaccag ctgtcaccat cccagactgg aagatcacat
2161 ccagcacaca cagccactcc ccagatgacc tctgcagcat caaacctcct gcctcctgtt
2221 gcatttaaaa acacaattgg acgcatagta ctttcggaac ataaaagtgt caaatttaat
2281 tgctcgatca acattcctaa cgtgtatcaa gaaacagctg gcatttcgtg gtggaaagat
2341 ggaaaggaac tgcttggggc acatcattca atcacacagt tttatcctga tgaggaaggg
2401 gtatcaataa ttgcattgtt caggtatgtg tccttccttt ctaaatgcca tcgtgctaat
2461 accccctcca aagcagaatc cccaaggaaa gagcaccctg gttttgtatg tatgattgtt
2521 cgtcaagcaa tcttgatcct tttagagggt agatacatag ctgacctggc tgtgttgttg
2581 atttctttta aatgagcttg tattgtggta actgctggct aacacacgct ctgagggttt
2641 agagggctgc tcccagctca tctaaggtga gtattaagta caccttagat ccaatggcta
2701 ccagagagtg ggaaacagag tgtatgaagg gggtgtgttc atgtttgtgc ctgtgcgtgt
2761 gtgcgccagc cccgttgcag tggtcctcgc tagctgtcag tgggcagagc gcacatcgcc
2821 cacagtcccc gagaagttgg ggggaggggt cggcaattga acgggtgcct agagaaggtg
2881 gcgcggggta aactgggaaa gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg
2941 gggagaaccg tatataagtg cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc
3001 cgccagaaca cagctgaagc ttcgaggggc tcgcatctct ccttcacgcg cccgccgccc
3061 tacctgaggc cgccatccac gccggttgag tcgcgttctg ccgcctcccg cctgtggtgc
3121 ctcctgaact gcgtccgccg tctaggtaag tttaaagctc aggtcgagac cgggcctttg
3181 tccggcgctc ccttggagcc tacctagact cagccggctc tccacgcttt gcctgaccct
3241 gcttgctcaa ctctacgtct ttgtttcgtt ttctgttctg cgccgttaca gatccaagct
3301 gtgaccggcg cgtcgacgcc accatggtga gcaagggcga ggagctgttc accggggtgg
3361 tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg
3421 agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca
3481 agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg cagtgcttca
3541 gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct
3601 acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg
3661 tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg
3721 aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata
3781 tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg
3841 aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc
3901 ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc aaagacccca
3961 acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg
4021 gcatggacga gctgtacaag tccggactca gatctcgaga ggaggaggag gagacagaca
4081 gcaggatgcc ccacctcgac agccccggca gctcccagcc gagacgctcc ttcctctcaa
4141 gggtgatcag ggcagcgcta ccgttgcagc tgcttctgct gctgctgctg ctcctggcct
4201 gcctgctacc tgcctctgaa gatgactaca gctgcaccca ggccaacaac tttgcccgat
4261 ccttctaccc catgctgcgg tacaccaacg ggccacctcc cacctaggaa ttcaataaaa
4321 gatctttatt ttcattagat ctgtgtgttg gttttttgtg tgcacgtgcg gaccgagcgg
4381 ccgcaggaac ccctagtgat ggagttggcc actccctctc tgcgcgctcg ctcgctcact
4441 gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc
4501 gagcgagcgc gcagctgcct gcaggggcgc ctgatgcggt attttctcct tacgcatctg
4561 tgcggtattt cacaccgcat acgtcaaagc aaccatagta cgcgccctgt agcggcgcat
4621 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccttag
4681 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
4741 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
4801 ccaaaaaact tgatttgggt gatggttcac gtagtgggcc atcgccctga tagacggttt
4861 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
4921 caacactcaa ctctatctcg ggctattctt ttgatttata agggattttg ccgatttcgg
4981 tctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
5041 taacgtttac aattttatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
5101 gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg
5161 catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac
5221 cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta
5281 atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg
5341 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
5401 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
5461 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
5521 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
5581 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
5641 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
5701 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
5761 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
5821 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
5881 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
5941 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
6001 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
6061 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
6121 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
6181 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
6241 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
6301 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
6361 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
6421 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
6481 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
6541 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
6601 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
6661 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
6721 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
6781 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
6841 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
6901 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
6961 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
7021 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
7081 ttttacggtt cctggccttt tgctggcctt ttgctcacat gt
//
