| 质粒名称: | pRC_RR_VP1_r1c3 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 65724 |
| 存储实验室: | Junghae Suh |
|
载体骨架基本信息 |
|
| 载体名称: | AAV2_G453_delGTTTQSR |
| 载体出品公司: | -- |
| 空载体大小: | -- |
| 完整质粒大小: | 7975 bp |
| 载体修饰: | -- |
| 载体类型: | AAV |
| 筛选标记: | -- |
| 载体抗性: | 氨苄青霉素，100μg/mL |
| 生长温度: | 37 ℃ |
| 宿主菌: | NEB Stable |
| 拷贝数: | -- |
|
插入基因信息 |
|
| 插入基因名称: | mCherry |
| 物种来源: | -- |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子: | -- |
| 标签/融合蛋白: | -- |
| 克隆方法: | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pRC_RR_VP1_r1c3 was a gift from Junghae Suh (Addgene plasmid # 65724 ; http://n2t.net/addgene:65724 ; RRID:Addgene_65724) |
| 参考文献部分: | Random Insertion of mCherry Into VP3 Domain of Adeno-associated Virus Yields Fluorescent Capsids With no Loss of Infectivity. Judd J, Wei F, Nguyen PQ, Tartaglia LJ, Agbandje-McKenna M, Silberg JJ, Suh J. Mol Ther Nucleic Acids. 2012 Nov 13;1:e54. doi: 10.1038/mtna.2012.46. 10.1038/mtna.2012.46 PubMed 23629029 |
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 pRC_RR_VP1_r1c3 7975 bp ds-DNA circular SYN 10-JAN-2022
DEFINITION Rescued infectious variant from pAAV2_RaPID library; mCherry
insertion in place of the deleted amino acid sequence 453-GTTTQSR.
ACCESSION .
VERSION .
KEYWORDS pRC_RR_VP1_r1c3
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7975)
AUTHORS Judd J, Wei F, Nguyen PQ, Tartaglia LJ, Agbandje-McKenna M, Silberg
JJ, Suh J
TITLE Random Insertion of mCherry Into VP3 Domain of Adeno-associated
Virus Yields Fluorescent Capsids With no Loss of Infectivity.
JOURNAL Mol Ther Nucleic Acids. 2012 Nov 13;1:e54. doi:
10.1038/mtna.2012.46.
PUBMED 23629029
REFERENCE 2 (bases 1 to 7975)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Jan 10, 2022 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..7975
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 1..17
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 27..46
/label=T7
/note="T7 promoter, forward primer"
promoter 27..45
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind 70..86
/label=pBluescriptKS
/note="For pBluescript vector"
primer_bind 71..87
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(121..137)
/label=SK primer
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(121..137)
/label=pBluescriptSK
/note="For pBluescript vector"
CDS 3512..4219
/codon_start=1
/product="monomeric derivative of DsRed fluorescent protein
(Shaner et al., 2004)"
/label=mCherry
/note="mammalian codon-optimized"
/translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
EGRHSTGGMDELYK"
primer_bind complement(3657..3675)
/label=mCherry-R
/note="mCherry, reverse primer"
primer_bind 3925..3944
/label=mCherry-F
/note="mCherry, forward primer"
CDS 4898..4909
/codon_start=1
/product="WELQut protease recognition and cleavage site"
/label=WELQut site
/translation="WELQ"
primer_bind complement(5188..5208)
/label=T3
/note="T3 promoter, forward primer"
promoter complement(5188..5206)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(5227..5243)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(5227..5243)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(5240..5262)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 5251..5267
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
promoter complement(5275..5305)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 5320..5341
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(5458..5475)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(5629..6217)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(5709..5728)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(6388..7248)
/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 7011..7030
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(7249..7353)
/gene="bla"
/label=AmpR promoter
rep_origin complement(7379..7834)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(7516..7537)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 7728..7747
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 7961..8
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 7975..17
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
ORIGIN
1 gtaaaacgac ggccagtgag cgcgcgtaat acgactcact atagggcgaa ttgggtaccg
61 ggccccccct cgaggtcgac ggtatcgata agcttgatat cgaattcctg cagcccgggg
121 gatccactag ttctagagtc ctgtattaga ggtcacgtga gtgttttgcg acattttgcg
181 acaccatgtg gtcacgctgg gtatttaagc ccgagtgagc acgcagggtc tccattttga
241 agcgggaggt ttgaacgcgc agccgccatg ccggggtttt acgagattgt gattaaggtc
301 cccagcgacc ttgacgagca tctgcccggc atttctgaca gctttgtgaa ctgggtggcc
361 gagaaggaat gggagttgcc gccagattct gacatggatc tgaatctgat tgagcaggca
421 cccctgaccg tggccgagaa gctgcagcgc gactttctga cggaatggcg tcgtgtgagt
481 aaggccccgg aggccctttt ctttgtgcaa tttgagaagg gagagagcta cttccacatg
541 cacgtgctcg tggaaaccac cggggtgaaa tctatggttt tgggacgttt cctgagtcag
601 attcgcgaaa aactgattca gagaatttac cgcgggatcg agccgacttt gccaaactgg
661 ttcgcggtca caaagaccag aaatggtgcc ggaggcggga acaaggtggt ggatgagtgc
721 tacatcccca attacttgct ccccaaaacc cagcctgagc tccagtgggc gtggactaat
781 atggaacagt atttaagcgc ctgtttgaat ctcacggagc gtaaacggtt ggtggcgcag
841 catctgacgc acgtgtcgca gacgcaggag cagaacaaag agaatcagaa tcccaattct
901 gatgcgccgg tgatcagatc aaaaacttca gccaggtaca tggagctggt cgggtggctc
961 gtggacaagg ggattacctc ggagaagcag tggatacagg aggaccaggc ctcatacatc
1021 tccttcaatg cggcctccaa ctcgcggtcc caaatcaagg ctgccttgga caatgcggga
1081 aagattatga gcctgactaa aaccgccccc gactacctgg tgggccagca gcccgtggag
1141 gacatttcca gcaatcggat ttataaaatt ttggaactaa acgggtacga tccccaatat
1201 gcggcttccg tctttctggg atgggccacg aaaaagttcg gcaagaggaa caccatctgg
1261 ctgtttgggc ctgcaactac cgggaagacc aacatcgcgg aggccatagc ccacactgtg
1321 cccttctacg ggtgcgtaaa ctggaccaat gagaactttc ccttcaacga ctgtgtcgac
1381 aagatggtga tctggtggga ggaggggaag atgaccgcca aggtcgtgga gtcggccaaa
1441 gccattctcg gaggaagcaa ggtgcgcgtg gaccagaaat gcaagtcctc ggcccagata
1501 gacccgactc ccgtgatcgt cacctccaac accaacatgt gcgccgtgat tgacgggaac
1561 tcaacgacct tcgaacacca gcagccgttg caagaccgga tgttcaaatt tgaactcacc
1621 cgccgtctgg atcatgactt tgggaaggtc accaagcagg aagtcaaaga ctttttccgg
1681 tgggcaaagg atcacgtggt tgaggtggag catgaattct acgtcaaaaa gggtggagcc
1741 aagaaaagac ccgcccccag tgacgcagat ataagtgagc ccaaacgggt gcgcgagtca
1801 gttgcgcagc catcgacgtc agacgcggaa gcttcgatca actacgcaga caggtaccaa
1861 aacaaatgtt ctcgtcacgt gggcatgaat ctgatgctgt ttccctgcag acaatgcgag
1921 agaatgaatc agaattcaaa tatctgcttc actcacggac agaaagactg tttagagtgc
1981 tttcccgtgt cagaatctca acccgtttct gtcgtcaaaa aggcgtatca gaaactgtgc
2041 tacattcatc atatcatggg aaaggtgcca gacgcttgca ctgcctgcga tctggtcaat
2101 gtggatttgg atgactgcat ctttgaacaa taaatgattt aaatcaggta tggctgccga
2161 tggttatcta cccgattggc tcgaggacac tctctctgaa ggaataagac agtggtggaa
2221 gctcaaacct ggcccaccac caccaaagcc cgcagagcgg cataaggacg acagcagggg
2281 tcttgtgctt cctgggtaca agtacctcgg acccttcaac ggactcgaca agggagagcc
2341 ggtcaacgag gcagacgccg cggccctcga gcacgacaaa gcctacgacc ggcagctcga
2401 cagcggagac aacccgtacc tcaagtacaa ccacgccgac gcggagtttc aggagcgcct
2461 taaagaagat acgtcttttg ggggcaacct cggacgagca gtcttccagg cgaaaaagag
2521 ggttcttgaa cctctgggcc tggttgagga acctgttaag acggctccgg gaaaaaagag
2581 gccggtagag cactctcctg tggagccaga ctcctcctcg ggaaccggaa aggcgggcca
2641 gcagcctgca agaaaaagat tgaattttgg tcagactgga gacgcagact cagtacctga
2701 cccccagcct ctcggacagc caccagcagc cccctctggt ctgggaacta ataccatggc
2761 tacaggcagt ggcgcaccaa tggcagacaa taacgagggt gccgacggag tgggtaattc
2821 ctcgggaaat tggcattgcg attccacatg gatgggcgac agagtcatca ccaccagcac
2881 ccgaacctgg gccctgccca cctacaacaa ccacctctac aaacaaattt ccagccaatc
2941 aggagcctcg aacgacaatc actactttgg ctacagcacc ccttgggggt attttgactt
3001 caacagattc cactgccact tttcaccacg tgactggcaa agactcatca acaacaactg
3061 gggattccga cccaagagac tcaacttcaa gctctttaac attcaagtca aagaggtcac
3121 gcagaatgac ggtacgacga cgattgccaa taaccttacc agcacggttc aggtgtttac
3181 tgactcggag taccagctcc cgtacgtcct cggctcggcg catcaaggat gcctcccgcc
3241 gttcccagca gacgtcttca tggtgccaca gtatggatac ctcaccctga acaacgggag
3301 tcaggcagta ggacgctctt cattttactg cctggagtac tttccttctc agatgctgcg
3361 taccggaaac aactttacct tcagctacac ttttgaggac gttcctttcc acagcagcta
3421 cgctcacagc cagagtctgg accgtctcat gaatcctctc atcgaccagt acctgtatta
3481 cttgagcaga acaaacactc caagtgccgg catggtgagc aagggcgagg aggataacat
3541 ggccatcatc aaggagttca tgcgcttcaa ggtgcacatg gagggctccg tgaacggcca
3601 cgagttcgag atcgagggcg agggcgaggg ccgcccctac gagggcaccc agaccgccaa
3661 gctgaaggtg accaagggtg gccccctgcc cttcgcctgg gacatcctgt cccctcagtt
3721 catgtacggc tccaaggcct acgtgaagca ccccgccgac atccccgact acttgaagct
3781 gtccttcccc gagggcttca agtgggagcg cgtgatgaac ttcgaggacg gcggcgtggt
3841 gaccgtgacc caggactcct ccctgcagga cggcgagttc atctacaagg tgaagctgcg
3901 cggcaccaac ttcccctccg acggccccgt aatgcagaag aagactatgg gctgggaggc
3961 ctcctccgag cggatgtacc ccgaggacgg tgccctgaag ggcgagatca agcagaggct
4021 gaagctgaag gacggcggcc actacgacgc tgaggtcaag accacctaca aggccaagaa
4081 gcccgtgcag ctgcccggcg cttacaacgt caacatcaag ttggacatca cctcccacaa
4141 cgaggactac accatcgtgg aacagtacga acgcgccgag ggccgccact ccaccggcgg
4201 catggacgag ctgtacaagg gcgcccttca gttttctcag gccggagcga gtgacattcg
4261 ggaccagtct aggaactggc ttcctggacc ctgttaccgc cagcagcgag tatcaaagac
4321 atctgcggat aacaacaaca gtgaatactc gtggactgga gctaccaagt accacctcaa
4381 tggcagagac tctctggtga atccgggccc ggctatggca agccacaagg acgatgaaga
4441 aaagtttttt cctcagagcg gggttctcat ctttgggaag caaggctcag agaaaacaaa
4501 tgtggacatt gaaaaggtca tgattacaga cgaagaggaa atcaggacaa ccaatcccgt
4561 ggctacggag cagtatggtt ctgtatctac caacctccag agaggcaaca gacaagcagc
4621 taccgcagat gtcaacacac aaggcgttct tccaggcatg gtctggcagg acagagatgt
4681 gtaccttcag gggcccatct gggcaaagat tccacacacg gacggacatt ttcacccctc
4741 tcccctcatg ggtggattcg gacttaaaca ccctcctcca cagattctca tcaagaacac
4801 cccggtgcct gcgaatcctt cgaccacctt cagtgcggca aagtttgctt ccttcatcac
4861 acagtactcc acgggacagg tcagcgtgga gatcgagtgg gagctgcaga aggaaaacag
4921 caaacgctgg aatcccgaaa ttcagtacac ttccaactac aacaagtctg ttaatgtgga
4981 ctttactgtg gacactaatg gcgtgtattc agagcctcgc cccattggca ccagatacct
5041 gactcgtaat ctgtaattgg caggtcttgt taatcaataa accgtttaat tcgtttcagt
5101 tgaactttgg tctctgcgta tttctttctt atctagtttc catgctctag agcggccgcc
5161 accgcggtgg agctccagct tttgttccct ttagtgaggg ttaattgcgc gcttggcgta
5221 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
5281 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
5341 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
5401 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
5461 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
5521 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
5581 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
5641 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
5701 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
5761 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
5821 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
5881 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
5941 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
6001 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
6061 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
6121 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
6181 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
6241 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
6301 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
6361 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
6421 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
6481 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
6541 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
6601 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
6661 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
6721 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
6781 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
6841 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
6901 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
6961 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
7021 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
7081 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
7141 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
7201 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
7261 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
7321 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctaa
7381 attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt
7441 tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata
7501 gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac
7561 gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa
7621 tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc
7681 cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg
7741 aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca
7801 cccgccgcgc ttaatgcgcc gctacagggc gcgtcccatt cgccattcag gctgcgcaac
7861 tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga
7921 tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgtt
//
