| 质粒名称: | AAV pEF1a-DIO-FLPo-WPRE-hGHpA |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 87306 |
| 存储实验室: | Li Zhang |
|
载体骨架基本信息 |
|
| 载体名称: | pAAV |
| 载体出品公司: | Stratagene |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 5601 bp |
| 完整质粒大小: | 6892 bp |
| 载体修饰: | 由Addgene质粒20297和51669修饰而成。 |
| 载体类型: | 哺乳动物表达,AAV |
| 高拷贝/低拷贝: | 高拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | Cre-dependent mouse codon-optimized Flp recombinase,DIO-FLPo |
| 物种来源: | M. musculus (mouse), S. cerevisiae (budding yeast), Synthetic |
| 插入大小: | 1299 bp |
| 启动子 | EF1a |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | NheI(未破坏) |
| 3' 克隆位点: | AscI(未破坏) |
| 5' 测序引物 | AGCTGACAGGTGGTGGCAAT |
| 3' 测序引物 | TCAAGCCTCAGACAGTGGTTC |
|
文献引用方法 |
|
| 材料和方法部分: | AAV pEF1a-DIO-FLPo-WPRE-hGHpA was a gift from Li Zhang (Addgene plasmid # 87306 ; http://n2t.net/addgene:87306 ; RRID:Addgene_87306) |
| 参考文献部分: | AAV-Mediated Anterograde Transsynaptic Tagging: Mapping Corticocollicular Input-Defined Neural Pathways for Defense Behaviors. Zingg B, Chou XL, Zhang ZG, Mesik L, Liang F, Tao HW, Zhang LI. Neuron. 2017 Jan 4;93(1):33-47. doi: 10.1016/j.neuron.2016.11.045. Epub 2016 Dec 15. 10.1016/j.neuron.2016.11.045 PubMed 27989459 |
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 pEF1a-DIO-FLPo-WPRE-hGHpA 6892 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION An AAV vector expresses FLPo in a Cre recombinase-dependent manner..
ACCESSION .
VERSION .
KEYWORDS AAV pEF1a-DIO-FLPo-WPRE-hGHpA
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6892)
AUTHORS Zingg B, Chou XL, Zhang ZG, Mesik L, Liang F, Tao HW, Zhang LI
TITLE AAV-Mediated Anterograde Transsynaptic Tagging: Mapping
Corticocollicular Input-Defined Neural Pathways for Defense
Behaviors.
JOURNAL Neuron. 2017 Jan 4;93(1):33-47. doi: 10.1016/j.neuron.2016.11.045.
Epub 2016 Dec 15.
PUBMED 27989459
REFERENCE 2 (bases 1 to 6892)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6892
/organism="synthetic DNA construct"
/mol_type="other DNA"
repeat_region 13..142
/label=AAV2 ITR (alternate)
/note="Functional equivalent of wild-type AAV2 ITR"
promoter 243..1421
/label=EF-1-alpha promoter
/note="strong constitutive promoter for human elongation
factor EF-1-alpha"
intron 474..1412
/label=EF-1-alpha intron A
/note="intron upstream of the start codon of human
EF-1-alpha"
primer_bind 1369..1389
/label=EF1a-F
/note="Human elongation factor-1a promoter, forward primer"
protein_bind 1452..1485
/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 1536..1569
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT)."
CDS complement(1581..2879)
/codon_start=1
/product="nuclear-targeted site-specific recombinase"
/label=FLPo
/note="FLPo is a mouse codon-optimized fusion between the
SV40 nuclear localization signal and a thermostable version
of the Saccharomyces cerevisiae site-specific recombinase
FLP. Recombination occurs at FRT sequences."
/translation="MAPKKKRKVMSQFDILCKTPPKVLVRQFVERFERPSGEKIASCAA
ELTYLCWMITHNGTAIKRATFMSYNTIISNSLSFDIVNKSLQFKYKTQKATILEASLKK
LIPAWEFTIIPYNGQKHQSDITDIVSSLQLQFESSEEADKGNSHSKKMLKALLSEGESI
WEITEKILNSFEYTSRFTKTKTLYQFLFLATFINCGRFSDIKNVDPKSFKLVQNKYLGV
IIQCLVTETKTSVSRHIYFFSARGRIDPLVYLDEFLRNSEPVLKRVNRTGNSSSNKQEY
QLLKDNLVRSYNKALKKNAPYPIFAIKNGPKSHIGRHLMTSFLSMKGLTELTNVVGNWS
DKRASAVARTTYTHQITAIPDHYFALVSRYYAYDPISKEMIALKDETNPIEEWQHIEQL
KGSAEGSIRYPAWNGIISQEVLDYLSSYINRRI"
protein_bind complement(2889..2922)
/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(2973..3006)
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT)."
misc_feature 3031..3619
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(3084..3104)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(3502..3513)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
polyA_signal 3651..4127
/label=hGH poly(A) signal
/note="human growth hormone polyadenylation signal"
primer_bind complement(3850..3869)
/label=hGH-PA-R
/note="Human growth hormone terminator, reverse primer"
repeat_region 4167..4307
/label=AAV2 ITR
/note="inverted terminal repeat of adeno-associated virus
serotype 2"
repeat_region 4167..4296
/label=AAV2 ITR
rep_origin 4382..4837
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(4469..4488)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 4679..4700
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(4854..4873)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 4973..4995
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(5033..5051)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 5119..5223
/gene="bla"
/label=AmpR promoter
CDS 5224..6084
/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(5442..5461)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 6255..6843
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 6744..6763
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 ttgctcacat gtcctgcagg cagctgcgcg ctcgctcgct cactgaggcc gcccgggcgt
61 cgggcgacct ttggtcgccc ggcctcagtg agcgagcgag cgcgcagaga gggagtggcc
121 aactccatca ctaggggttc ctgcggccgc acgcgtaagc tttgcaaaga tggataaagt
181 tttaaacaga gaggaatctt tgcagctaat ggaccttcta ggtcttgaaa ggagtgggaa
241 ttggctccgg tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc gagaagttgg
301 ggggaggggt cggcaattga accggtgcct agagaaggtg gcgcggggta aactgggaaa
361 gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg gggagaaccg tatataagtg
421 cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca caggtaagtg
481 ccgtgtgtgg ttcccgcggg cctggcctct ttacgggtta tggcccttgc gtgccttgaa
541 ttacttccac ctggctgcag tacgtgattc ttgatcccga gcttcgggtt ggaagtgggt
601 gggagagttc gaggccttgc gcttaaggag ccccttcgcc tcgtgcttga gttgaggcct
661 ggcctgggcg ctggggccgc cgcgtgcgaa tctggtggca ccttcgcgcc tgtctcgctg
721 ctttcgataa gtctctagcc atttaaaatt tttgatgacc tgctgcgacg ctttttttct
781 ggcaagatag tcttgtaaat gcgggccaag atctgcacac tggtatttcg gtttttgggg
841 ccgcgggcgg cgacggggcc cgtgcgtccc agcgcacatg ttcggcgagg cggggcctgc
901 gagcgcggcc accgagaatc ggacgggggt agtctcaagc tggccggcct gctctggtgc
961 ctggcctcgc gccgccgtgt atcgccccgc cctgggcggc aaggctggcc cggtcggcac
1021 cagttgcgtg agcggaaaga tggccgcttc ccggccctgc tgcagggagc tcaaaatgga
1081 ggacgcggcg ctcgggagag cgggcgggtg agtcacccac acaaaggaaa agggcctttc
1141 cgtcctcagc cgtcgcttca tgtgactcca cggagtaccg ggcgccgtcc aggcacctcg
1201 attagttctc gagcttttgg agtacgtcgt ctttaggttg gggggagggg ttttatgcga
1261 tggagtttcc ccacactgag tgggtggaga ctgaagttag gccagcttgg cacttgatgt
1321 aattctcctt ggaatttgcc ctttttgagt ttggatcttg gttcattctc aagcctcaga
1381 cagtggttca aagttttttt cttccatttc aggtgtcgtg aggtaccgga tcctctagag
1441 tcgactccgg aataacttcg tataggatac tttatacgaa gttatgcaga atggtagctg
1501 gattgtagct gctattagca atatgaaacc tcttaataac ttcgtatagc atacattata
1561 cgaagttatg gcgcgcctta tcagatccgc ctgttgatgt agctgctcag gtagtccagc
1621 acctcctggc tgatgatgcc gttccaggcg gggtatctga tgctgccctc ggcgctgccc
1681 ttcagctgct cgatgtgctg ccactcctcg atggggttgg tctcgtcctt cagggcgatc
1741 atctccttgc tgatggggtc gtaggcgtag tacctggaca ccagggcgaa gtagtggtcg
1801 gggatggcgg tgatctggtg ggtgtaggtg gtcctggcca cggcggaggc cctcttgtcg
1861 ctccagttgc ccaccacgtt tgtcagctcg gtcaggccct tcatgctcag aaagctggtc
1921 atcaggtgcc tgccgatgtg gctcttaggg ccgttcttga tagcgaagat ggggtagggg
1981 gcgttcttct tcagggcctt gttgtagctg cgcaccaggt tgtccttcag cagctggtac
2041 tcctgcttgt tgctgctgct gttgccggtc ctgttcactc tcttcagcac gggctcgctg
2101 ttcctcagga actcgtccag gtacaccagg gggtcgatcc tgcctctggc gctgaaaaag
2161 tagatgtgcc tggacacgct tgtcttggtc tcggtcacca ggcactgaat gatcacgccc
2221 aggtacttgt tctgcaccag cttgaagctc ttggggtcca cgttcttgat gtcgctgaac
2281 ctgccgcagt tgatgaatgt ggccaggaac aggaactggt acagggtctt ggtcttggtg
2341 aacctgctgg tgtactcgaa gctgttcagg atcttctcgg tgatctccca gatgctctcg
2401 ccctcggaca gcagggcctt cagcatcttc ttgctgtggc tgttgccctt gtcggcctcc
2461 tcgctgctct cgaactgcag ctgcaggctg gacacgatgt cggtgatgtc gctctggtgc
2521 ttctggccgt tgtaagggat gatggtgaac tcccaggcgg ggatcagctt cttcaggctg
2581 gcctccagga tggtggcctt ctgggtcttg tacttgaact gcaggctctt gttcacgatg
2641 tcgaagctca ggctgttgct gatgatggtg ttgtagctca tgaaggtggc cctcttgatg
2701 gcggtgccgt tgtgggtgat catccagcac aggtaggtca gctcggcggc acagctggcg
2761 atcttctcgc cgctgggcct ctcgaatctc tccacgaact gccgcaccag caccttgggg
2821 ggggtcttgc acaggatgtc gaactggctc atcaccttcc tcttcttctt aggagccatg
2881 gtgctagcat aacttcgtat aaagtatcct atacgaagtt atttgcctta acccagaaat
2941 tatcactgtt attctttaga atggtgcaaa gaataacttc gtataatgta tgctatacga
3001 agttatgaat tcgatatcaa gcttatcgat aatcaacctc tggattacaa aatttgtgaa
3061 agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta
3121 atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa
3181 tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg
3241 tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc
3301 ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc
3361 cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg
3421 gggaaatcat cgtcctttcc ttggctgctc gcctatgttg ccacctggat tctgcgcggg
3481 acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg
3541 ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc
3601 ctttgggccg cctccccgca tcgataccga gcgctgctcg agagatctac gggtggcatc
3661 cctgtgaccc ctccccagtg cctctcctgg ccctggaagt tgccactcca gtgcccacca
3721 gccttgtcct aataaaatta agttgcatca ttttgtctga ctaggtgtcc ttctataata
3781 ttatggggtg gaggggggtg gtatggagca aggggcaagt tgggaagaca acctgtaggg
3841 cctgcggggt ctattgggaa ccaagctgga gtgcagtggc acaatcttgg ctcactgcaa
3901 tctccgcctc ctgggttcaa gcgattctcc tgcctcagcc tcccgagttg ttgggattcc
3961 aggcatgcat gaccaggctc agctaatttt tgtttttttg gtagagacgg ggtttcacca
4021 tattggccag gctggtctcc aactcctaat ctcaggtgat ctacccacct tggcctccca
4081 aattgctggg attacaggcg tgaaccactg ctcccttccc tgtccttctg attttgtagg
4141 taaccacgtg cggaccgagc ggccgcagga acccctagtg atggagttgg ccactccctc
4201 tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt
4261 tgcccgggcg gcctcagtga gcgagcgagc gcgcagctgc ctgcaggggc gcctgatgcg
4321 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag
4381 tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
4441 gctacacttg ccagcgcctt agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
4501 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
4561 agtgctttac ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg
4621 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
4681 ggactcttgt tccaaactgg aacaacactc aactctatct cgggctattc ttttgattta
4741 taagggattt tgccgatttc ggtctattgg ttaaaaaatg agctgattta acaaaaattt
4801 aacgcgaatt ttaacaaaat attaacgttt acaattttat ggtgcactct cagtacaatc
4861 tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc
4921 tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc
4981 tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg
5041 atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc
5101 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
5161 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
5221 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
5281 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
5341 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
5401 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
5461 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
5521 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
5581 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
5641 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
5701 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
5761 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
5821 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
5881 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
5941 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
6001 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
6061 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
6121 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
6181 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
6241 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
6301 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
6361 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
6421 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
6481 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
6541 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
6601 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
6661 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
6721 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
6781 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
6841 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc tt
//
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