| 质粒名称: | pAAV-EF1a-double floxed-EYFP-WPRE-HGHpA |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 20296 |
| 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: | 6324 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: | EYFP |
| 物种来源: | A. victoria |
| 插入大小: | 720 bp |
| 基因又名: | enhanced yellow fluorescent protein；YFP |
| 标签/融合蛋白: | 人细胞色素c氧化酶亚基VIII（N端） |
| 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-EYFP-WPRE-HGHpA was a gift from Karl Deisseroth (爱迪基因 plasmid # 20296) |
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-EYFP-WPRE-HGHpA 6324 bp ds-DNA circular SYN 10-JAN-2018
DEFINITION Cre-activated AAV expression of EYFP.
ACCESSION .
VERSION .
KEYWORDS pAAV-EF1a-double floxed-EYFP-WPRE-HGHpA
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6324)
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-EYFP-WPRE-HGHpA|
FEATURES Location/Qualifiers
source 1..6324
/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(1579..2298)
/codon_start=1
/product="enhanced YFP"
/label=EYFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(1624..1645)
/label=EGFP-C
/note="EGFP, forward primer"
primer_bind 1973..1992
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 2232..2253
/label=EGFP-N
/note="EGFP, reverse primer"
regulatory 2295..2304
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
protein_bind complement(2309..2342)
/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(2393..2426)
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT)."
misc_feature 2451..3039
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(2504..2524)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(2922..2933)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
polyA_signal 3071..3547
/label=hGH poly(A) signal
/note="human growth hormone polyadenylation signal"
primer_bind complement(3270..3289)
/label=hGH-PA-R
/note="Human growth hormone terminator, reverse primer"
repeat_region 3587..3727
/label=AAV2 ITR
/note="inverted terminal repeat of adeno-associated virus
serotype 2"
repeat_region 3587..3716
/label=AAV2 ITR
rep_origin 3802..4257
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(3889..3908)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 4099..4120
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(4274..4293)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 4393..4415
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(4453..4471)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 4539..4643
/gene="bla"
/label=AmpR promoter
CDS 4644..5504
/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(4862..4881)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 5675..6263
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 6164..6183
/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 gcgccctatt acttgtacag ctcgtccatg ccgagagtga tcccggcggc
1621 ggtcacgaac tccagcagga ccatgtgatc gcgcttctcg ttggggtctt tgctcagggc
1681 ggactggtag ctcaggtagt ggttgtcggg cagcagcacg gggccgtcgc cgatgggggt
1741 gttctgctgg tagtggtcgg cgagctgcac gctgccgtcc tcgatgttgt ggcggatctt
1801 gaagttcacc ttgatgccgt tcttctgctt gtcggccatg atatagacgt tgtggctgtt
1861 gtagttgtac tccagcttgt gccccaggat gttgccgtcc tccttgaagt cgatgccctt
1921 cagctcgatg cggttcacca gggtgtcgcc ctcgaacttc acctcggcgc gggtcttgta
1981 gttgccgtcg tccttgaaga agatggtgcg ctcctggacg tagccttcgg gcatggcgga
2041 cttgaagaag tcgtgctgct tcatgtggtc ggggtagcgg gcgaagcact gcaggccgta
2101 gccgaaggtg gtcacgaggg tgggccaggg cacgggcagc ttgccggtgg tgcagatgaa
2161 cttcagggtc agcttgccgt aggtggcatc gccctcgccc tcgccggaca cgctgaactt
2221 gtggccgttt acgtcgccgt ccagctcgac caggatgggc accaccccgg tgaacagctc
2281 ctcgcccttg ctcaccatgg tggctagcat aacttcgtat aaagtatcct atacgaagtt
2341 atttgcctta acccagaaat tatcactgtt attctttaga atggtgcaaa gaataacttc
2401 gtataatgta tgctatacga agttatgaat tcgatatcaa gcttatcgat aatcaacctc
2461 tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc
2521 tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca
2581 ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg
2641 tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca
2701 ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg
2761 cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg
2821 acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc gcctatgttg
2881 ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg
2941 accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc
3001 ctcagacgag tcggatctcc ctttgggccg cctccccgca tcgataccga gcgctgctcg
3061 agagatctac gggtggcatc cctgtgaccc ctccccagtg cctctcctgg ccctggaagt
3121 tgccactcca gtgcccacca gccttgtcct aataaaatta agttgcatca ttttgtctga
3181 ctaggtgtcc ttctataata ttatggggtg gaggggggtg gtatggagca aggggcaagt
3241 tgggaagaca acctgtaggg cctgcggggt ctattgggaa ccaagctgga gtgcagtggc
3301 acaatcttgg ctcactgcaa tctccgcctc ctgggttcaa gcgattctcc tgcctcagcc
3361 tcccgagttg ttgggattcc aggcatgcat gaccaggctc agctaatttt tgtttttttg
3421 gtagagacgg ggtttcacca tattggccag gctggtctcc aactcctaat ctcaggtgat
3481 ctacccacct tggcctccca aattgctggg attacaggcg tgaaccactg ctcccttccc
3541 tgtccttctg attttgtagg taaccacgtg cggaccgagc ggccgcagga acccctagtg
3601 atggagttgg ccactccctc tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag
3661 gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc gcgcagctgc
3721 ctgcaggggc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc
3781 atacgtcaaa gcaaccatag tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
3841 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
3901 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
3961 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgatttgg
4021 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
4081 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
4141 cgggctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
4201 agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaattttat
4261 ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc
4321 caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag
4381 ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg
4441 cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg
4501 tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
4561 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
4621 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
4681 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
4741 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
4801 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
4861 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
4921 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
4981 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
5041 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
5101 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
5161 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
5221 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
5281 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
5341 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
5401 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
5461 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
5521 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
5581 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
5641 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
5701 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
5761 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
5821 tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
5881 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
5941 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
6001 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
6061 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
6121 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
6181 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
6241 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
6301 tttgctggcc ttttgctcac atgt
//
