| 质粒名称: | AAV-6P-SEW-YC3.6 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 31379 |
| 存储实验室: | Fritjof Helmchen |
|
载体骨架基本信息 |
|
| 载体名称: | AAV-6P-SEWB |
| 载体出品公司: | -- |
| 空载体大小: | 4800 bp |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | AAV |
| 筛选标记: | -- |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | NEB Stable |
| 拷贝数: | 低拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | Yellow Cameleon 3.60,YC3.60 |
| 物种来源: | synthetic construct |
| 插入大小: | 1962 bp |
| 基因突变: | -- |
| GenBank编号 | AB178712 |
| 启动子: | -- |
| 标签/融合蛋白: |
eCFP（插件上的N端子）
Venus（插件上的C端子）
|
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | NheI (未破坏) |
| 3’克隆位点: | HindIII (未破坏) |
| 5’测序引物 | -- |
| 3’测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | AAV-6P-SEW-YC3.6 was a gift from Fritjof Helmchen (Addgene plasmid # 31379 ; http://n2t.net/addgene:31379 ; RRID:Addgene_31379) |
| 参考文献部分: | Chronic imaging of cortical sensory map dynamics using a genetically encoded calcium indicator. Minderer M, Liu W, Sumanovski LT, Kugler S, Helmchen F, Margolis DJ. J Physiol. 2011 Nov 14. 10.1113/jphysiol.2011.219014 PubMed 22083602 |
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 AAV-6P-SEW-YC3.6 6714 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS AAV-6P-SEW-YC3.6
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6714)
AUTHORS Minderer M, Liu W, Sumanovski LT, Kugler S, Helmchen F, Margolis DJ
TITLE Chronic imaging of cortical sensory map dynamics using a genetically
encoded calcium indicator.
JOURNAL J Physiol. 2011 Nov 14.
PUBMED 22083602
REFERENCE 2 (bases 1 to 6714)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6714
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 27..474
/label=hSyn promoter
/note="human synapsin I promoter; confers neuron-specific
expression (Kugler et al., 2003)"
regulatory 500..509
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 1646..1723
/codon_start=1
/product="calmodulin-binding peptide"
/label=CBP
/note="derived from skeletal muscle myosin light chain
kinase; binds calmodulin with nanomolar affinity in the
presence of calcium"
/translation="KRRWKKNFIAVSAANRFKKISSSGAL"
CDS 1730..2464
/codon_start=1
/product="circularly permuted form of the Venus yellow
fluorescent protein (Nagai et al., 2004)"
/label=cp173Venus
/note="efficient FRET acceptor for CFP"
/translation="MDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRD
HMVLLEFVTAAGITLGMDELYKGGSGGMVSKGEELFTGVVPILVELDGDVNGHKFSVSG
EGEGDATYGKLTLKLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPE
GYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHN
VYITADKQKNGIKANFKIRHNIE"
misc_feature 2487..3075
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(2540..2560)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(2958..2969)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
primer_bind complement(3111..3128)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 3117..3324
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 3578..4033
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(3665..3684)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 3875..3896
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(4100..4119)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 4219..4241
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(4279..4297)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 4365..4469
/gene="bla"
/label=AmpR promoter
CDS 4470..5330
/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(4688..4707)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 5501..6089
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 5990..6009
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 6243..6260
/label=L4440
/note="L4440 vector, forward primer"
polyA_signal 6558..6606
/note="synthetic polyadenylation signal"
misc_feature 6620..6711
/label=pause site
/note="RNA polymerase II transcriptional pause signal from
the human alpha-2 globin gene"
primer_bind 6660..6679
/label=RVprimer3
/note="pGL3 vector, forward primer"
ORIGIN
1 ctagactgca gagggccctg cgtatgagtg caagtgggtt ttaggaccag gatgaggcgg
61 ggtgggggtg cctacctgac gaccgacccc gacccactgg acaagcaccc aacccccatt
121 ccccaaattg cgcatcccct atcagagagg gggaggggaa acaggatgcg gcgaggcgcg
181 tgcgcactgc cagcttcagc accgcggaca gtgccttcgc ccccgcctgg cggcgcgcgc
241 caccgccgcc tcagcactga aggcgcgctg acgtcactcg ccggtccccc gcaaactccc
301 cttcccggcc accttggtcg cgtccgcgcc gccgccggcc cagccggacc gcaccacgcg
361 aggcgcgaga taggggggca cgggcgcgac catctgcgct gcggcgccgg cgactcagcg
421 ctgcctcagt ctgcggtggg cagcggagga gtcgtgtcgt gcctgagagc gcagtcgaat
481 tcaagctgct agcgcaagtg ccaccatggt gagcaagggc gaggagctgt tcaccggggt
541 ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaggttca gcgtgtccgg
601 cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg
661 caagctgccc gtgccctggc ccaccctcgt gaccaccctg acctggggcg tgcagtgctt
721 cagccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg
781 ctacgtccag gagcgtacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga
841 ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa
901 ggaggacggc aacatcctgg ggcacaagct ggagtacaac tatatcagcc acaacgtcta
961 tatcaccgcc gacaagcaga agaacggcat caaggccaac ttcaagatcc gccacaacat
1021 cgaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg
1081 ccccgtgctg ctgcccgaca accactacct gagcacccag tccgccctga gcaaagaccc
1141 caaggagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccc gcatgcatga
1201 ccaactgaca gaagagcaga ttgcagagtt caaagaagcc ttctcattat tcgacaagga
1261 tggggacggc accatcacca caaaggaact tggcaccgtt atgaggtcgc ttggacaaaa
1321 cccaacggaa gcagaattgc aggatatgat caatgaagtc gatgctgatg gcaatggaac
1381 gatttacttt cctgaatttc ttactatgat ggctagaaaa atgaaggaca cagacagcga
1441 agaggaaatc cgagaagcat tccgtgtttt tgacaaggat gggaacggct acatcagcgc
1501 tgctcagtta cgtcacgtca tgacaaacct cggggagaag ttaacagatg aagaagttga
1561 tgaaatgata agggaagcag atatcgatgg tgatggccaa gtaaactatg aagagtttgt
1621 acaaatgatg acagcaaagg gggggaagag gcgctggaag aaaaacttca ttgccgtcag
1681 cgctgccaac cggttcaaga agatctccag ctccggggca ctggagctca tggacggcgg
1741 cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct
1801 gcccgacaac cactacctga gctaccagtc cgccctgagc aaagacccca acgagaagcg
1861 cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga
1921 gctgtacaag ggtggcagcg gtggcatggt gagcaagggc gaggagctgt tcaccggggt
1981 ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg
2041 cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagctgatct gcaccaccgg
2101 caagctgccc gtgccctggc ccaccctcgt gaccaccctg ggctacggcc tgcagtgctt
2161 cgcccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg
2221 ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga
2281 ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa
2341 ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta
2401 tatcaccgcc gacaagcaga agaacggcat caaggccaac ttcaagatcc gccacaacat
2461 cgagtaagag gcgcaagctt atcgataatc aacctctgga ttacaaaatt tgtgaaagat
2521 tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc
2581 ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct
2641 ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca
2701 ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt
2761 ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg
2821 cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga
2881 agctgacgtc ctttccatgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt
2941 ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc
3001 cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt
3061 gggccgcctc cccgcctgat cgataccgtc gactagagct cgctgatcag cctcgactgt
3121 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
3181 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
3241 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga
3301 agacaatagc aggcatgctg gggagagatc taggaacccc tagtgatgga gttggccact
3361 ccctctctgc gcgctcgctc gctcactgag gccgcccggg caaagcccgg gcgtcgggcg
3421 acctttggtc gcccggcctc agtgagcgag cgagcgcgca gagagggagt ggccaacccc
3481 cccccccccc cccctgcagc cagctggcgt aatagcgaag aggcccgcac cgatcgccct
3541 tcccaacagt tgcgtagcct gaatggcgaa tggcgcgacg cgccctgtag cggcgcatta
3601 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
3661 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
3721 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
3781 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
3841 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
3901 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
3961 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
4021 acgtttacaa tttcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc
4081 gcatatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac
4141 acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca
4201 gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga
4261 aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa
4321 taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
4381 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
4441 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
4501 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
4561 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
4621 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
4681 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
4741 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
4801 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
4861 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
4921 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
4981 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
5041 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
5101 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
5161 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
5221 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
5281 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
5341 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
5401 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
5461 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
5521 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
5581 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
5641 atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
5701 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
5761 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
5821 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
5881 tacagcgtga gcattgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
5941 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
6001 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
6061 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
6121 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
6181 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
6241 gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg
6301 cgcgttggcc gattcattaa tgcagctggg ctgcaggggg gggggggggg gggttggcca
6361 ctccctctct gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacgcc
6421 cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg cagagaggga gtggccaact
6481 ccatcactag gggttcctag atctaggatc acgcgttcta gaaatattaa ggtacgggag
6541 gtacttggag cggccgcaat aaaatatctt tattttcatt acatctgtgt gttggttttt
6601 tgtgtgaatc gatagtacta acatacgctc tccatcaaaa caaaacgaaa caaaacaaac
6661 tagcaaaata ggctgtcccc agtgcaagtg caggtgccag aacatttctc tatt
//
