| 质粒名称: | rAAV-syn::FLEX-rev::PSAML141F,Y115F:GlyR-IRES-GFP |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 32481 |
| 存储实验室: | Scott Sternson |
|
载体骨架基本信息 |
|
| 载体名称: | AAV2 |
| 载体抗性: | 氨苄霉素 |
| 空载体大小: | 5100 bp |
| 完整质粒大小: | 7743 bp |
| 载体类型: | AAV, Cre/Lox ; Adeno-associated virus, FLEX switch |
| 高拷贝/低拷贝: | 低拷贝 |
| 生长条件: | 30 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | PSAML141F,Y115F-GlyR |
| 物种来源: | 人源 |
| 插入大小: | 2643 bp |
| 启动子 | Synapsin |
| 标签/融合蛋白: | IRES-EGFP (C terminal on insert) |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | speI blunt(未破坏) |
| 3' 克隆位点: | SpeI blunt(未破坏) |
| 5' 测序引物 | gtaagtatcaaggttacaag |
| 3' 测序引物 | ctgacaacgggccacaactc |
|
文献引用方法 |
|
| 材料和方法部分: | rAAV-syn::FLEX-rev::PSAML141F,Y115F:GlyR-IRES-GFP was a gift from Scott Sternson (爱迪基因 plasmid # 32481) |
| 参考文献部分: | Chemical and genetic engineering of selective ion channel-ligand interactions. Magnus CJ, Lee PH, Atasoy D, Su HH, Looger LL, Sternson SM. Science. 2011 Sep 2;333(6047):1292-6. 10.1126/science.1206606 PubMed 21885782 |
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rAAV-syn::FLEX-rev::PSAML141F,Y115F:GlyR-IRES-GFP载体质粒基本信息
rAAV-syn::FLEX-rev::PSAML141F,Y115F:GlyR-IRES-GFP质粒图谱载体图谱和rAAV-syn::FLEX-rev::PSAML141F,Y115F:GlyR-IRES-GFP载体序列质粒序列多克隆位点信息
rAAV-syn::FLEX-rev::PSAML141F,Y115F:GlyR-IRES-GFP质粒载体简介
rAAV-syn::FLEX-rev::PSAML141F,Y115F:GlyR-IRES-GFP质粒序列载体序列
LOCUS rAAV-syn::FLEX-rev::PSAML141F,Y115F:GlyR-IRES-GFP 7454 bp ds-DNA circular SYN 10-JAN-2018
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS rAAV-syn::FLEX-rev::PSAML141F,Y115F:GlyR-IRES-GFP
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7454)
AUTHORS Magnus CJ, Lee PH, Atasoy D, Su HH, Looger LL, Sternson SM
TITLE Chemical and genetic engineering of selective ion channel-ligand
interactions.
JOURNAL Science. 2011 Sep 2;333(6047):1292-6.
PUBMED 21885782
COMMENT This file is created by Vector NTI
http://www.biofeng.com/
COMMENT ORIGDB|GenBank
COMMENT VNTAUTHORNAME|biofeng.com|
COMMENT VNTNAME|rAAV-syn::FLEX-rev::PSAML141F,Y115F:GlyR-IRES-GFP|
FEATURES Location/Qualifiers
source 1..7454
/organism="synthetic DNA construct"
/mol_type="other DNA"
repeat_region 55..184
/label=AAV2 ITR
intron 777..909
/label=chimeric intron
/note="chimera between introns from human beta-globin and
immunoglobulin heavy chain genes"
primer_bind 892..911
/label=pMT2-F
/note="Synthetic intron, forward primer"
protein_bind complement(922..955)
/label=lox2272
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GGATACTT). lox2272 sites are compatible with each
other, but incompatible with loxP or loxN sites."
protein_bind complement(1018..1051)
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT)."
CDS complement(1062..1778)
/codon_start=1
/product="enhanced GFP with monomerizing A206K mutation
(Zacharias et al., 2002)"
/label=mEGFP
/note="mammalian codon-optimized"
/translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE
FVTAAGITLGMDELYK"
primer_bind complement(1107..1128)
/label=EGFP-C
/note="EGFP, forward primer"
primer_bind 1456..1475
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 1715..1736
/label=EGFP-N
/note="EGFP, reverse primer"
misc_feature 1802..2353
/label=IRES
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
primer_bind 2192..2209
/label=IRES reverse
/note="IRES internal ribosome entry site, reverse primer.
Also called pCDH-rev"
protein_bind 3708..3741
/label=lox2272
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GGATACTT). lox2272 sites are compatible with each
other, but incompatible with loxP or loxN sites."
protein_bind 3812..3845
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT)."
misc_feature 3917..4505
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(3970..3990)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(4388..4399)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
polyA_signal 4583..4704
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(4594..4613)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 4648..4667
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
repeat_region 4866..4995
/label=AAV2 ITR
primer_bind complement(5061..5078)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(5232..5820)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(5312..5331)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5991..6851)
/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 6614..6633
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(6852..6956)
/gene="bla"
/label=AmpR promoter
rep_origin complement(6982..7437)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(7119..7140)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 7331..7350
/label=F1ori-R
/note="F1 origin, reverse primer"
ORIGIN
1 ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctgcgc
61 gctcgctcgc tcactgaggc cgcccgggca aagcccgggc gtcgggcgac ctttggtcgc
121 ccggcctcag tgagcgagcg agcgcgcaga gagggagtgg ccaactccat cactaggggt
181 tccttgtagt taatgattaa cccgccatgc tacttatcta cgtagccatg ctctaggaag
241 atccgagctc cagtgtgctg gaattcgccc ttttcagtat ttaaattgac gaccgacccc
301 gacccactgg acaagcaccc aacccccatt ccccaaattg cgcatcccct atcagagagg
361 gggaggggaa acaggatgcg gcgaggcgcg tgcgcactgc cagcttcagc accgcggaca
421 gtgccttcgc ccccgcctgg cggcgcgcgc caccgccgcc tcagcactga aggcgcgctg
481 acgtcactcg ccggtccccc gcaaactccc cttcccggcc accttggtcg cgtccgcgcc
541 gccgccggcc cagccggacc gcaccacgcg aggcgcgaga taggggggca cgggcgcgac
601 catctgcgct gcggcgccgg cgactcagcg ctgcctcagt ctgcggtggg cagcggagga
661 gtcgtgtcgt gcctgagagc gcagctgtgc tcctgggcac cgcgcagtcc gcccccgcgg
721 ctcctggcca gaccacccct aggaccccct gccccaagtc gcagccgttg gatcaggtaa
781 gtatcaaggt tacaagacag gtttaaggag accaatagaa actgggcttg tcgagacaga
841 gaagactctt gcgtttctga taggcaccta ttggtcttac tgacatccac tttgcctttc
901 tctccacagg tgatgatccc cataacttcg tataaagtat cctatacgaa gttatatcaa
961 aataggaaga ccaatgcttc accatcgacc cgaattgcca agcatcacca tcgacacata
1021 acttcgtata atgtatgcta tacgaagtta tactagctag attacttgta cagctcgtcc
1081 atgccgagag tgatcccggc ggcggtcacg aactccagca ggaccatgtg atcgcgcttc
1141 tcgttggggt ctttgctcag cttggactgg gtgctcaggt agtggttgtc gggcagcagc
1201 acggggccgt cgccgatggg ggtgttctgc tggtagtggt cggcgagctg cacgctgccg
1261 tcctcgatgt tgtggcggat cttgaagttc accttgatgc cgttcttctg cttgtcggcc
1321 atgatataga cgttgtggct gttgtagttg tactccagct tgtgccccag gatgttgccg
1381 tcctccttga agtcgatgcc cttcagctcg atgcggttca ccagggtgtc gccctcgaac
1441 ttcacctcgg cgcgggtctt gtagttgccg tcgtccttga agaagatggt gcgctcctgg
1501 acgtagcctt cgggcatggc ggacttgaag aagtcgtgct gcttcatgtg gtcggggtag
1561 cggctgaagc actgcacgcc gtaggtcagg gtggtcacga gggtgggcca gggcacgggc
1621 agcttgccgg tggtgcagat gaacttcagg gtcagcttgc cgtaggtggc atcgccctcg
1681 ccctcgccgg acacgctgaa cttgtggccg tttacgtcgc cgtccagctc gaccaggatg
1741 ggcaccaccc cggtgaacag ctcctcgccc ttgctcaccg gatcccccat ggttgtggcc
1801 atattatcat cgtgtttttc aaaggaaaac cacgtccccg tggttcgggg ggcctagacg
1861 tttttttaac ctcgactaaa cacatgtaaa gcatgtgcac cgaggcccca gatcagatcc
1921 catacaatgg ggtaccttct gggcatcctt cagccccttg ttgaatacgc ttaggagagc
1981 catttgactc tttccacaac tatccaactc acaacgtggc actggggttg tgccgccttt
2041 gcaggtgtat cttatacacg tggcttttgg ccgcagaggc acctgtcgcc aggtgggggg
2101 ttccgctgcc tgcaaagggt cgctacagac gttgtttgtc ttcaagaagc ttccagagga
2161 actgcttcct tcacgacatt caacagacct tgcattcctt tggcgagagg ggaaagaccc
2221 ctaggaatgc tcgtcaagaa gacagggcca ggtttccggg ccctcacatt gccaaaagac
2281 ggcaatatgg tggaaaataa catatagaca aacgcacacc ggccttattc caagcggctt
2341 cggccagtaa cgttaggggg gggggcgggg ggaattgcgg ccgcttactg gttgtgtacg
2401 tcctcccttc gcacaatctt gtagatgatc caataaaaca tgttgaaaat caggaaagcc
2461 atggggaatc cgatccggga gattttgtcg atctttttcg ctctctgaat aaagagtttg
2521 cgcatttcct ccgggctctt agatggagca gggggtgggt tggttgtgtt gctattgttg
2581 gctcccttca cggaaattcc gtctttagcc tggaggcaag caggtcccat accataggcg
2641 ctaaagttga agcgtccttc tcctgcctca tcctctttgt ggtggcgtcg ttttcttctg
2701 aaccgcaaca gctctttatg ctgtcgagac acaaaattga cagcggcgta ttccagcagt
2761 gcgctgaaga caaacagaag gcagacagcc atccagatat cgatggcctt cacgtaagaa
2821 accttgggca agctggctcg gctgcctgag ctctgagtag tcattgtcag cactgtggtg
2881 atgcccagtc cgaccctagc tggagcggcg tccatattaa tccagaagga gatccatgac
2941 aaaatcacta taagcaagct tgggatatac atttggatca gataataacc catgcgtctc
3001 cgcatggtca ctgtgaaggt gacatctggg tagggctctt tgcagcattc ataaaatcgc
3061 tcgcttcttt taccggggat tcccacaagg tcccactccc cattcgggat gtaaccagag
3121 atgtcagcct cctgcatttg cagatccagg ctccatccgc cgtagctcca gctcccgaat
3181 ttgagtttgc agtgctggac gtcgaaggga aaccagcgga cgtcgatata gcatgaggac
3241 ttgaagattc caggggggaa gtactggcag tggcctgaag aattgaccaa cacgttggtg
3301 tgaaatgttg catcaaacct ttcgtctgcg ctgttgaaca gcaggatatc tggtttccag
3361 atttggccgt cagggaatct tacagtcttt acacctgggt attcgctcac attccactgc
3421 aagtagtgat cggtccagga catctgcaac caaatattag tggtcagcac ctggttcttt
3481 tcatccacgt ccatgatctg cagcaaggag aggctgaagt acactgtcag tggctgggaa
3541 tcatttgcga ccggccgctc cagggggtta taattcttaa caagctcctt atacagtttg
3601 cgctggaact caccctgcag gctaacgtgc agaagggaag cagccagggc gagccacacg
3661 ccgcctggag aacagcgcat ggtgaattca agcttgctag ccctagtata acttcgtata
3721 ggatacttta tacgaagtta tcattgggat tcttcctatt ttgatccaag catcaccatc
3781 gaccctctag tccagatctc accatcgacc cataacttcg tatagcatac attatacgaa
3841 gttatgtccc tcgaagaggt tcgaaacact gaaaagggcg aattctgcag atgatccact
3901 cgagaagctt atcgataatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat
3961 tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca
4021 tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc
4081 tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc
4141 tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt
4201 cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg
4261 gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc
4321 ctttccttgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta
4381 cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg
4441 gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc
4501 cccgcatcga taccgtcgac ccgggcggcc gctcgagcat gcatctagag tcgacccggg
4561 cggccgcttc gagcagacat gataagatac attgatgagt ttggacaaac cacaactaga
4621 atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc
4681 attataagct gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt
4741 cagggggaga tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg tggtaaaatc
4801 gataaggatc ttcctagagc atggctacgt agataagtag catggcgggt taatcattaa
4861 ctacaaggaa cccctagtga tggagttggc cactccctct ctgcgcgctc gctcgctcac
4921 tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag
4981 cgagcgagcg cgcagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
5041 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
5101 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
5161 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
5221 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
5281 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
5341 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
5401 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
5461 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
5521 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
5581 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
5641 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
5701 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
5761 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
5821 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
5881 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
5941 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
6001 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
6061 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
6121 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
6181 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
6241 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
6301 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
6361 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
6421 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
6481 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
6541 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
6601 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
6661 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
6721 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
6781 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
6841 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
6901 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
6961 tttccccgaa aagtgccacc taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa
7021 atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata
7081 aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac
7141 tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc
7201 cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa
7261 atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg
7321 cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg
7381 tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtccc
7441 attcgccatt cagg
//
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