| 质粒名称: | pAAV-EF1a-double floxed-mCherry-WPRE-HGHpA |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 20299 |
| 存储实验室: | Karl Deisseroth |
|
载体骨架基本信息 |
|
| 载体名称: | pAAV |
| 载体出品公司: | Stratagene |
| 载体抗性: | 氨苄霉素 |
| 空载体大小: | 5604 bp |
| 完整质粒大小: | 6315 bp |
| 载体类型: | AAV,哺乳细胞表达 |
| 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: | mCherry |
| 插入大小: | 711 bp |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | AscI(未破坏) |
| 3' 克隆位点: | NheI(未破坏) |
| 5' 测序引物 | n/a |
|
文献引用方法 |
|
| 材料和方法部分: | pAAV-EF1a-double floxed-mCherry-WPRE-HGHpA was a gift from Karl Deisseroth (爱迪基因 plasmid # 20299) |
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 pAAV-EF1a-double floxed-mCherry-WPRE-HGHpA 6312 bp ds-DNA circular SYN 10-JAN-2018
DEFINITION Cre-activated AAV expression of mCherry.
ACCESSION .
VERSION .
KEYWORDS pAAV-EF1a-double floxed-mCherry-WPRE-HGHpA
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6312)
TITLE Cre-activated AAV expression plasmids
COMMENT This file is created by Vector NTI
http://www.biofeng.com/
COMMENT ORIGDB|GenBank
COMMENT VNTAUTHORNAME|biofeng.com|
COMMENT VNTNAME|pAAV-EF1a-double floxed-mCherry-WPRE-HGHpA|
FEATURES Location/Qualifiers
source 1..6312
/organism="synthetic DNA construct"
/mol_type="other DNA"
repeat_region 1..141
/label=AAV2 ITR
/note="inverted terminal repeat of adeno-associated virus
serotype 2"
repeat_region 12..141
/label=AAV2 ITR
promoter 242..1419
/label=EF-1-alpha promoter
/note="strong constitutive promoter for human elongation
factor EF-1-alpha"
intron 473..1410
/label=EF-1-alpha intron A
/note="intron upstream of the start codon of human
EF-1-alpha"
primer_bind 1367..1387
/label=EF1a-F
/note="Human elongation factor-1a promoter, forward primer"
protein_bind 1450..1483
/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 1534..1567
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT)."
CDS complement(1576..2286)
/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(1854..1873)
/label=mCherry-F
/note="mCherry, forward primer"
primer_bind 2123..2141
/label=mCherry-R
/note="mCherry, reverse primer"
regulatory 2283..2292
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
protein_bind complement(2297..2330)
/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(2381..2414)
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT)."
misc_feature 2439..3027
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(2492..2512)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(2910..2921)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
polyA_signal 3059..3535
/label=hGH poly(A) signal
/note="human growth hormone polyadenylation signal"
primer_bind complement(3258..3277)
/label=hGH-PA-R
/note="Human growth hormone terminator, reverse primer"
repeat_region 3575..3715
/label=AAV2 ITR
/note="inverted terminal repeat of adeno-associated virus
serotype 2"
repeat_region 3575..3704
/label=AAV2 ITR
rep_origin 3790..4245
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(3877..3896)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 4087..4108
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(4262..4281)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 4381..4403
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(4441..4459)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 4527..4631
/gene="bla"
/label=AmpR promoter
CDS 4632..5492
/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(4850..4869)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 5663..6251
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 6152..6171
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
121 actccatcac taggggttcc tgcggccgca cgcgtaagct ttgcaaagat ggataaagtt
181 ttaaacagag aggaatcttt gcagctaatg gaccttctag gtcttgaaag gagtgggaat
241 tggctccggt gcccgtcagt gggcagagcg cacatcgccc acagtccccg agaagttggg
301 gggaggggtc ggcaattgaa ccggtgccta gagaaggtgg cgcggggtaa actgggaaag
361 tgatgtcgtg tactggctcc gcctttttcc cgagggtggg ggagaaccgt atataagtgc
421 agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac aggtaagtgc
481 cgtgtgtggt tcccgcgggc ctggcctctt tacgggttat ggcccttgcg tgccttgaat
541 tacttccact ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg
601 gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg
661 cctgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct
721 ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg
781 caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc
841 gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga
901 gcgcggccac cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct
961 ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca
1021 gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg
1081 acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg
1141 tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat
1201 tagttctcga gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg
1261 gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa
1321 ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca
1381 gtggttcaaa gtttttttct tccatttcag gtgtcgtgag gtaccggatc ctctagagtc
1441 gactccggaa taacttcgta taggatactt tatacgaagt tatgcagaat ggtagctgga
1501 ttgtagctgc tattagcaat atgaaacctc ttaataactt cgtatagcat acattatacg
1561 aagttatggc gcgccttact tgtacagctc gtccatgccg ccggtggagt ggcggccctc
1621 ggcgcgttcg tactgttcca cgatggtgta gtcctcgttg tgggaggtga tgtccaactt
1681 gatgttgacg ttgtaggcgc cgggcagctg cacgggcttc ttggccttgt aggtggtctt
1741 gacctcagcg tcgtagtggc cgccgtcctt cagcttcagc ctctgcttga tctcgccctt
1801 cagggcgccg tcctcggggt acatccgctc ggaggaggcc tcccagccca tggtcttctt
1861 ctgcattacg gggccgtcgg aggggaagtt ggtgccgcgc agcttcacct tgtagatgaa
1921 ctcgccgtcc tgcagggagg agtcctgggt cacggtcacc acgccgccgt cctcgaagtt
1981 catcacgcgc tcccacttga agccctcggg gaaggacagc ttcaagtagt cggggatgtc
2041 ggcggggtgc ttcacgtagg ccttggagcc gtacatgaac tgaggggaca ggatgtccca
2101 ggcgaagggc agggggccac ccttggtcac cttcagcttg gcggtctggg tgccctcgta
2161 ggggcggccc tcgccctcgc cctcgatctc gaactcgtgg ccgttcacgg agccctccat
2221 gtgcaccttg aagcgcatga actccttgat gatggccatg ttatcctcct cgcccttgct
2281 caccatggtg gctagcataa cttcgtataa agtatcctat acgaagttat ttgccttaac
2341 ccagaaatta tcactgttat tctttagaat ggtgcaaaga ataacttcgt ataatgtatg
2401 ctatacgaag ttatgaattc gatatcaagc ttatcgataa tcaacctctg gattacaaaa
2461 tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg
2521 ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct
2581 tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg
2641 gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct
2701 gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg
2761 ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg
2821 tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctatgttgcc acctggattc
2881 tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc
2941 gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc
3001 ggatctccct ttgggccgcc tccccgcatc gataccgagc gctgctcgag agatctacgg
3061 gtggcatccc tgtgacccct ccccagtgcc tctcctggcc ctggaagttg ccactccagt
3121 gcccaccagc cttgtcctaa taaaattaag ttgcatcatt ttgtctgact aggtgtcctt
3181 ctataatatt atggggtgga ggggggtggt atggagcaag gggcaagttg ggaagacaac
3241 ctgtagggcc tgcggggtct attgggaacc aagctggagt gcagtggcac aatcttggct
3301 cactgcaatc tccgcctcct gggttcaagc gattctcctg cctcagcctc ccgagttgtt
3361 gggattccag gcatgcatga ccaggctcag ctaatttttg tttttttggt agagacgggg
3421 tttcaccata ttggccaggc tggtctccaa ctcctaatct caggtgatct acccaccttg
3481 gcctcccaaa ttgctgggat tacaggcgtg aaccactgct cccttccctg tccttctgat
3541 tttgtaggta accacgtgcg gaccgagcgg ccgcaggaac ccctagtgat ggagttggcc
3601 actccctctc tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc
3661 ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc gcagctgcct gcaggggcgc
3721 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat acgtcaaagc
3781 aaccatagta cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
3841 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
3901 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
3961 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgatttgggt gatggttcac
4021 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
4081 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg ggctattctt
4141 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
4201 aaaaatttaa cgcgaatttt aacaaaatat taacgtttac aattttatgg tgcactctca
4261 gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg
4321 acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct
4381 ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg
4441 gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt
4501 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
4561 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
4621 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
4681 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
4741 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
4801 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
4861 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
4921 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
4981 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
5041 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
5101 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
5161 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
5221 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
5281 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
5341 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
5401 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
5461 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
5521 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
5581 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
5641 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
5701 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
5761 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt
5821 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
5881 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
5941 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
6001 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
6061 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
6121 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
6181 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
6241 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
6301 ttgctcacat gt
//
