| 质粒名称: | pUCmini-iCAP-AAV.CAP-B22 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 175005 |
| 存储实验室: | Viviana Gradinaru |
|
载体骨架基本信息 |
|
| 载体名称: | pUC57-mini |
| 载体出品公司: | -- |
| 空载体大小: | 6311 bp |
| 完整质粒大小: | 8543 bp |
| 载体修饰: | -- |
| 载体类型: | 哺乳细胞表达, AAV |
| 筛选标记: | -- |
| 载体抗性: | 氨苄霉素100 μg/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: | Synthetic construct isolate AAV.CAP-B22 VP1 gene,AAV.CAP-B22 Cap |
| 物种来源: | Synthetic |
| 插入大小: | 2232 bp |
| 基因突变: | 7 amino acid substitution between VP1 452 and VP1 460 from NGSGQNQ to DGQSSKS |
| GenBank编号 | -- |
| 启动子: | p41 |
| 标签/融合蛋白: | -- |
| 克隆方法: | Gibson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: | pUCmini-iCAP-AAV.CAP-B22 was a gift from Viviana Gradinaru (Addgene plasmid # 175005 ; http://n2t.net/addgene:175005 ; RRID:Addgene_175005) |
| 参考文献部分: | AAV capsid variants with brain-wide transgene expression and decreased liver targeting after intravenous delivery in mouse and marmoset. Goertsen D, Flytzanis NC, Goeden N, Chuapoco MR, Cummins A, Chen Y, Fan Y, Zhang Q, Sharma J, Duan Y, Wang L, Feng G, Chen Y, Ip NY, Pickel J, Gradinaru V. Nat Neurosci. 2022 Jan;25(1):106-115. doi: 10.1038/s41593-021-00969-4. Epub 2021 Dec 9. 10.1038/s41593-021-00969-4 PubMed 34887588 |
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 pUCmini-iCAP-AAV.CAP-B22 8543 bp ds-DNA circular SYN 05-NOV-2021
DEFINITION non-standard AAV2 rep-AAV.CAP-B22 cap plasmid with AAV cap
expression controlled by a tTA-TRE amplifcation system.
ACCESSION .
VERSION .
KEYWORDS pUCmini-iCAP-AAV.CAP-B22
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8543)
TITLE Brain-wide gene expression with decreased liver targeting after
intravenous delivery of engineered AAV capsid in mouse and marmoset
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 8543)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Nov 5, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..8543
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 99..121
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 113..130
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 114..130
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 140..159
/label=T7
/note="T7 promoter, forward primer"
promoter 140..158
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 2748..2858
/codon_start=1
/product="3 tandem repeats of the minimal activation domain
of herpes simplex virus VP16 (Cheng et al., 2013)"
/label=VP48
/translation="DALDDFDLDMLPADALDDFDLDMLPADALDDFDLDML"
promoter complement(2897..2915)
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind complement(2898..2915)
/label=SP6
/note="SP6 promoter, forward primer"
primer_bind complement(2935..2952)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 2941..3165
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
protein_bind 3227..3245
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 3264..3282
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 3301..3319
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 3338..3356
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 3375..3393
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 3412..3430
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 3449..3467
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 3486..3504
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
CDS 6221..6232
/codon_start=1
/product="WELQut protease recognition and cleavage site"
/label=WELQut site
/translation="WELQ"
rep_origin complement(6788..7376)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(6868..6887)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(7547..8407)
/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 8170..8189
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(8408..8512)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 ccaatgatac gcgtcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg
61 tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac gttgtaaaac
121 gacggccagt gagcgcgcgt aatacgactc actatagggc gaattgggta catcgacggt
181 atcgggggag ctcgcagggt ctccattttg aagcgggagg tttgaacgcg cagccgccat
241 gccggggttt tacgagattg tgattaaggt ccccagcgac cttgacgagc atctgcccgg
301 catttctgac agctttgtga actgggtggc cgagaaggaa tgggagttgc cgccagattc
361 tgacatggat ctgaatctga ttgagcaggc acccctgacc gtggccgaga agctgcagcg
421 cgactttctg acggaatggc gccgtgtgag taaggccccg gaggctcttt tctttgtgca
481 atttgagaag ggagagagct acttccacat gcacgtgctc gtggaaacca ccggggtgaa
541 atccatggtt ttgggacgtt tcctgagtca gattcgcgaa aaactgattc agagaattta
601 ccgcgggatc gagccgactt tgccaaactg gttcgcggtc acaaagacca gaaatggcgc
661 cggaggcggg aacaaggtgg tggatgagtg ctacatcccc aattacttgc tccccaaaac
721 ccagcctgag ctccagtggg cgtggactaa tatggaacag tatttaagcg cctgtttgaa
781 tctcacggag cgtaaacggt tggtggcgca gcatctgacg cacgtgtcgc agacgcagga
841 gcagaacaaa gagaatcaga atcccaattc tgatgcgccg gtgatcagat caaaaacttc
901 agccaggtac atggagctgg tcgggtggct cgtggacaag gggattacct cggagaagca
961 gtggatccag gaggaccagg cctcatacat ctccttcaat gcggcctcca actcgcggtc
1021 ccaaatcaag gctgccttgg acaatgcggg aaagattatg agcctgacta aaaccgcccc
1081 cgactacctg gtgggccagc agcccgtgga ggacatttcc agcaatcgga tttataaaat
1141 tttggaacta aacgggtacg atccccaata tgcggcttcc gtctttctgg gatgggccac
1201 gaaaaagttc ggcaagagga acaccatctg gctgtttggg cctgcaacta ccgggaagac
1261 caacatcgcg gaggccatag cccacactgt gcccttctac gggtgcgtaa actggaccaa
1321 tgagaacttt cccttcaacg actgtgtgga caagatggtg atctggtggg aggaggggaa
1381 gatgaccgcc aaggtcgtgg agtcggccaa agccattctc ggaggaagca aggtgcgcgt
1441 ggaccagaaa tgcaagtcct cggcccagat agacccgact cccgtgatcg tcacctccaa
1501 caccaatatg tgcgccgtga ttgacgggaa ctcaacgacc ttcgaacacc agcagccgtt
1561 gcaagaccgg atgttcaaat ttgaactcac ccgccgtctg gatcatgact ttgggaaggt
1621 caccaagcag gaagtcaaag actttttccg gtgggcaaag gatcacgtgg ttgaggtgga
1681 gcatgaattc tacgtcaaaa agggtggagc caagaaaaga cccgccccca gtgacgcaga
1741 tataagtgag cccaaacggg tgcgcgagtc agttgcgcag ccatcgacgt cagacgcgga
1801 agcttcgatc aactacgcgg acaggtacca aaacaaatgt tctcgtcacg tgggcatgaa
1861 tctgatgctg tttccctgca gacaatgcga gagactgaat cagaattcaa atatctgctt
1921 cactcacggt gtcaaagact gtttagagtg ctttcccgtg tcagaatctc aacccgtttc
1981 tgtcgtcaaa aaggcgtatc agaaactgtg ctacattcat cacatcatgg gaaaggtgcc
2041 agacgcttgc actgcttgcg acctggtcaa tgtggacttg gatgactgtg tttctgaaca
2101 ataaatgact taaaccaggt atgagtcggc tggataaatc taaagtcata aacggcgctc
2161 tggaattact caatgaagtc ggtatcgaag gcctgacgac aaggaaactc gctcaaaagc
2221 tgggagttga gcagcctacc ctgtactggc acgtgaagaa caagcgggcc ctgctcgatg
2281 ccctggccat cgagatgctg gacaggcatc atacccactt ctgccccctg gaaggcgagt
2341 catggcaaga ctttctgcgg aacaacgcca agtcattccg ctgtgctctc ctctcacatc
2401 gcgacggggc taaagtgcat ctcggcaccc gcccaacaga gaaacagtac gaaaccctgg
2461 aaaatcagct cgcgttcctg tgtcagcaag gcttctccct ggagaacgca ctgtacgctc
2521 tgtccgccgt gggccacttt acactgggct gcgtattgga ggaacaggag catcaagtag
2581 caaaagagga aagagagaca cctaccaccg attctatgcc cccacttctg agacaagcaa
2641 ttgagctgtt cgaccggcag ggagccgaac ctgccttcct tttcggcctg gaactaatca
2701 tatgtggcct ggagaaacag ctaaagtgcg aaagcggcgg gccggccgac gcccttgacg
2761 attttgactt agacatgctc ccagccgatg cccttgacga ctttgacctt gatatgctgc
2821 ctgctgacgc tcttgacgat tttgaccttg acatgctccc cgggtaaatg catgaattcg
2881 atctagaggg ccctattcta tagtgtcacc taaatgctag agctcgctga tcagcctcga
2941 ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc
3001 tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc
3061 tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt
3121 gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa
3181 gaaccagctg gggctcgaat caagctatca agtgccacct gacgtctccc tatcagtgat
3241 agagaagtcg acacgtctcg agctccctat cagtgataga gaaggtacgt ctagaacgtc
3301 tccctatcag tgatagagaa gtcgacacgt ctcgagctcc ctatcagtga tagagaaggt
3361 acgtctagaa cgtctcccta tcagtgatag agaagtcgac acgtctcgag ctccctatca
3421 gtgatagaga aggtacgtct agaacgtctc cctatcagtg atagagaagt cgacacgtct
3481 cgagctccct atcagtgata gagaaggtac cccctatata agcagagaga tctgttcaaa
3541 tttgaactga ctaagcggct cccgccagat tttggcaaga ttactaagca ggaagtcaag
3601 gacttttttg cttgggcaaa ggtcaatcag gtgccggtga ctcacgagtt taaagttccc
3661 agggaattgg cgggaactaa aggggcggag aaatctctaa aacgcccact gggtgacgtc
3721 accaatacta gctataaaag tctggagaag cgggccaggc tctcatttgt tcccgagacg
3781 cctcgcagtt cagacgtgac tgttgatccc gctcctctgc gaccgctagc ttcgatcaac
3841 tacgcagaca ggtaccaaaa caagtgttct cgtcacgtgg gcattaatct gattctgttt
3901 ccctgcagac aatgcgagag aatgaatcag aactcaaata tctgcttcac tcacggacag
3961 aaagactgtt tagagtgctt tcccgtgtca gaatctcaac ccgtttctgt cgtcaaaaag
4021 gcgtatcaga aactgtgcta cattcatcat atcatgggaa aggtgccaga cgcttgcact
4081 gcctgcgatc tggtcaatgt ggatttggat gactgcatct ttgaacaata aatgacttaa
4141 gccaggtatg gctgccgatg gttatcttcc agattggctc gaggacaacc ttagtgaagg
4201 aattcgcgag tggtgggctt tgaaacctgg agcccctcaa cccaaggcaa atcaacaaca
4261 tcaagacaac gctagaggtc ttgtgcttcc gggttacaaa taccttggac ccggcaacgg
4321 actcgacaag ggggagccgg tcaacgcagc agacgcggcg gccctcgagc acgacaaagc
4381 ctacgaccag cagctcaagg ccggagacaa cccgtacctc aagtacaacc acgccgacgc
4441 cgagttccag gagcggctca aagaagatac gtcttttggg ggcaacctcg ggcgagcagt
4501 cttccaggcc aaaaagaggc ttcttgaacc tcttggtctg gttgaggaag cggctaagac
4561 ggctcctgga aagaagaggc ctgtagagca gtctcctcag gaaccggact cctccgcggg
4621 tattggcaaa tcgggtgcac agcccgctaa aaagagactc aatttcggtc agactggcga
4681 cacagagtca gtcccagacc ctcaaccaat cggagaacct cccgcagccc cctcaggtgt
4741 gggatctctt acaatggctt caggtggtgg cgcaccagtg gcagacaata acgaaggtgc
4801 cgatggagtg ggtagttcct cgggaaattg gcattgcgat tcccaatggc tgggggacag
4861 agtcatcacc accagcaccc gaacctgggc cctgcccacc tacaacaatc acctctacaa
4921 gcaaatctcc aacagcacat ctggaggatc ttcaaatgac aacgcctact tcggctacag
4981 caccccctgg gggtattttg acttcaacag attccactgc cacttctcac cacgtgactg
5041 gcagcgactc atcaacaaca actggggatt ccggcctaag cgactcaact tcaagctctt
5101 taacattcag gtcaaagagg ttacggacaa caatggagtc aagaccatcg ccaataacct
5161 taccagcacg gtccaggtct tcacggactc agactatcag ctcccgtacg tgctcgggtc
5221 ggctcacgag ggctgcctcc cgccgttccc agcggacgtt ttcatgattc ctcagtacgg
5281 gtatctgacg cttaatgatg gaagccaggc cgtgggtcgt tcgtcctttt actgcctgga
5341 atatttcccg tcgcaaatgc taagaacggg taacaacttc cagttcagct acgagtttga
5401 gaacgtacct ttccatagca gctacgctca cagccaaagc ctggaccgac taatgaatcc
5461 actcatcgac caatacttgt actatctctc tagaactatt gacggccaaa gcagcaaaag
5521 ccaaacgcta aaattcagtg tggccggacc cagcaacatg gctgtccagg gaagaaacta
5581 catacctgga cccagctacc gacaacaacg tgtctcaacc actgtgactc aaaacaacaa
5641 cagcgaattt gcttggcctg gagcttcttc ttgggctctc aatggacgta atagcttgat
5701 gaatcctgga cctgctatgg cctctcacaa agaaggagag gaccgtttct ttcctttgtc
5761 tggatcttta atttttggca aacaaggtac tggcagagac aacgtggatg cggacaaagt
5821 catgataacc aacgaagaag aaattaaaac tactaacccg gtagcaacgg agtcctatgg
5881 acaagtggcc acaaaccacc agagtgatgg gactttggcg gtgcctttta aggcacaggc
5941 gcagaccggt tgggttcaaa accaaggaat acttccgggt atggtttggc aggacagaga
6001 tgtgtacctg caaggaccca tttgggccaa aattcctcac acggacggca actttcaccc
6061 ttctccgctg atgggagggt ttggaatgaa gcacccgcct cctcagatcc tcatcaaaaa
6121 cacacctgta cctgcggatc ctccaacggc cttcaacaag gacaagctga actctttcat
6181 cacccagtat tctactggtc aagtcagcgt ggagatcgag tgggagctgc agaaggaaaa
6241 cagcaagcgc tggaacccgg agatccagta cacttccaac tattacaagt ctaataatgt
6301 tgaatttgct gttaatactg aaggtgtata tagtgaaccc cgccccattg gcaccagata
6361 cctgactcgt aatctgtaag tcgacttgct tgttaatcaa taaaccgttt aattcgtttc
6421 agttgaactt tggtctctgc gaagggcaat tcgtttaaac ctgcaggact agaggtcctg
6481 tattagaggt cacgtgagtg ttttgcgaca ttttgcgaca ccatgtggtc acgctgggta
6541 tttaagcccg agtgagcacg cagggtctcc attttgaagc gggaggtttg aacgcgcagc
6601 cgccaagccg aattctgcag atatcacatg tcctaggaac tatcgatcca tcacactggc
6661 ggccgctcga ctagagcggc cgccaccgcg gtggagctcc agcttttgcg gaccgaatcg
6721 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
6781 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
6841 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
6901 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
6961 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
7021 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
7081 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
7141 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
7201 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
7261 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
7321 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
7381 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
7441 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
7501 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
7561 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
7621 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
7681 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
7741 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
7801 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
7861 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
7921 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
7981 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
8041 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
8101 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
8161 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
8221 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
8281 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
8341 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
8401 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
8461 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
8521 cccgaaaagt gccacctgac gtc
//
