| 质粒名称: | pAAV-FLEX-GFP |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 28304 |
| 存储实验室: | Edward Boyden |
|
载体骨架基本信息 |
|
| 载体名称: | AAV-FLEX |
| 载体出品公司: | Scott Sternson |
| 载体抗性: | 氨苄霉素 |
| 空载体大小: | 5047 bp |
| 完整质粒大小: | 5767 bp |
| 载体类型: | 哺乳细胞表达，AAV, Cre/Lox ; Adeno-associated virus |
| 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: | GFP |
| 物种来源: | A. victoria |
| 插入大小: | 720 bp |
| 基因又名: | Green fluorescent protein |
| 基因突变: | N/A |
| 启动子 | CAG |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | KpnI(未破坏) |
| 3' 克隆位点: | BsrGI(未破坏) |
| 5' 测序引物 | cttctggcgtgtgaccgg |
|
文献引用方法 |
|
| 材料和方法部分: | pAAV-FLEX-GFP was a gift from Edward Boyden (爱迪基因 plasmid # 28304) |
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-FLEX-GFP 5764 bp ds-DNA circular SYN 10-JAN-2018
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pAAV-FLEX-GFP
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5764)
TITLE Accessory plasmids for optogenetics
JOURNAL Unpublished
COMMENT This file is created by Vector NTI
http://www.biofeng.com/
COMMENT ORIGDB|GenBank
COMMENT VNTAUTHORNAME|biofeng.com|
COMMENT VNTNAME|pAAV-FLEX-GFP|
FEATURES Location/Qualifiers
source 1..5764
/organism="synthetic DNA construct"
/mol_type="other DNA"
repeat_region 56..185
/label=AAV2 ITR
polyA_signal 347..468
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(384..403)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 438..457
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
misc_feature 511..1099
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
CDS 617..628
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
primer_bind 1026..1046
/label=WPRE-R
/note="WPRE, reverse primer"
protein_bind complement(1141..1174)
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT)."
protein_bind complement(1245..1278)
/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."
regulatory 1295..1304
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 1301..2020
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(1346..1367)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(1607..1626)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 1954..1975
/label=EGFP-C
/note="EGFP, forward primer"
protein_bind 2043..2076
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT)."
protein_bind 2139..2172
/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."
primer_bind complement(2207..2226)
/label=pCAG-F
/note="Rabbit beta-globin intron, for pCAG plasmids,
forward primer"
primer_bind complement(2288..2310)
/label=pCAGGS-5
/note="Chimeric intron in CAG promoter, forward primer"
promoter complement(2589..2866)
/label=chicken beta-actin promoter
repeat_region 3177..3306
/label=AAV2 ITR
rep_origin 3378..3833
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(3465..3484)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 3675..3696
/label=F1ori-F
/note="F1 origin, forward primer"
promoter 3859..3963
/gene="bla"
/label=AmpR promoter
CDS 3964..4824
/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(4182..4201)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 4995..5583
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 5484..5503
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 5737..5754
/label=L4440
/note="L4440 vector, forward primer"
ORIGIN
1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctgcg
61 cgctcgctcg ctcactgagg ccgcccgggc aaagcccggg cgtcgggcga cctttggtcg
121 cccggcctca gtgagcgagc gagcgcgcag agagggagtg gccaactcca tcactagggg
181 ttccttgtag ttaatgatta acccgccatg ctacttatct acgtagccat gctctaggaa
241 gatccttatc gattttacca catttgtaga ggttttactt gctttaaaaa acctcccaca
301 tctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc
361 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
421 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgctc
481 gaagcggccg cccgggtcga cggtatcgat gcggggaggc ggcccaaagg gagatccgac
541 tcgtctgagg gcgaaggcga agacgcggaa gaggccgcag agccggcagc aggccgcggg
601 aaggaaggtc cgctggattg agggccgaag ggacgtagca gaaggacgtc ccgcgcagaa
661 tccaggtggc aacacaggcg agcagccaag gaaaggacga tgatttcccc gacaacacca
721 cggaattgtc agtgcccaac agccgagccc ctgtccagca gcgggcaagg caggcggcga
781 tgagttccgc cgtggcaata gggaggggga aagcgaaagt cccggaaagg agctgacagg
841 tggtggcaat gccccaacca gtgggggttg cgtcagcaaa cacagtgcac accacgccac
901 gttgcctgac aacgggccac aactcctcat aaagagacag caaccaggat ttatacaagg
961 aggagaaaat gaaagccata cgggaagcaa tagcatgata caaaggcatt aaagcagcgt
1021 atccacatag cgtaaaagga gcaacatagt taagaatacc agtcaatctt tcacaaattt
1081 tgtaatccag aggttgatta tcgataagct tgatatcgaa ttcgaacctc ttcgagggac
1141 ataacttcgt ataatgtatg ctatacgaag ttatgggtcg atggtgagat ctggactaga
1201 gggtcgatgg tgatgcttgg atcaaaatag gaagaatccc aatgataact tcgtataaag
1261 tatcctatac gaagttatac tagaattcgg taccgccacc atggtgagca agggcgagga
1321 gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa
1381 gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt
1441 catttgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta
1501 cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc
1561 cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta
1621 caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa
1681 gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa
1741 cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa
1801 gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac
1861 ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccgc
1921 cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc
1981 cgccgggatc actctcggca tggacgagct gtacaagtaa aactcgaggg gggatcccta
2041 gtataacttc gtatagcata cattatacga agttatgggt cgatggtgat gcttggcaat
2101 tcgggtcgat ggtgaagcat tggtcttcct attttgatat aacttcgtat aggatacttt
2161 atacgaagtt atggggatcc aattctttgc caaaatgatg agacagcaca ataaccagca
2221 cgttgcccag gagctgtagg aaaaagaaga aggcatgaac atggttagca gaggctctag
2281 agccgccggt cacacgccag aagccgaacc ccgccctgcc ccgtcccccc cgaaggcagc
2341 cgtccccccg cggacagccc cgagccgctc cccccgcaca aagggccctc ccggagcccc
2401 tcaaggcttt cacgcagcca cagaaaagaa acaagccgtc attaaaccaa gcgctaatta
2461 cagcccggag gagaagggcc gtcccgcccg ctcacctgtg ggagtaacgc ggtcagtcag
2521 agccggggcg ggcggcgcga ggcggcggcg gagcggggca cggggcgaag gcagcgcgca
2581 gcgactcccg cccgccgcgc gcttcgcttt ttatagggcc gccgccgccg ccgcctcgcc
2641 ataaaaggaa actttcggag cgcgccgctc tgattggctg ccgccgcacc tctccgcctc
2701 gccccgcccc gcccctcgcc ccgccccgcc ccgcctggcg cgcgcccccc cccccccccc
2761 gcccccatcg ctgcacaaaa taattaaaaa ataaataaat acaaaattgg gggtggggag
2821 gggggggaga tggggagagt gaagcagaac gtggggctca cctcgaccat ggtaatagcg
2881 atgactaata cgtagatgta ctgccaagta ggaaagtccc ataaggtcat gtactgggca
2941 taatgccagg cgggccattt accgtcattg acgtcaatag ggggcgtact tggcatatga
3001 tacacttgat gtactgccaa gtgggcagtt taccgtaaat actccaccca ttgacgtcaa
3061 tggaaagtcc ctattggcgt tactatggga acatacgtca ttattgacgt caatggtact
3121 cttcctagag catggctacg tagataagta gcatggcggg ttaatcatta actacaagga
3181 acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca ctgaggccgg
3241 gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc
3301 gcgcagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag
3361 cctgaatggc gaatgggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
3421 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
3481 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
3541 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
3601 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
3661 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
3721 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
3781 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa tttaggtggc
3841 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
3901 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
3961 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
4021 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
4081 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
4141 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
4201 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
4261 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
4321 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
4381 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
4441 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
4501 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
4561 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
4621 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
4681 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
4741 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
4801 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
4861 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
4921 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
4981 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
5041 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
5101 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
5161 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
5221 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
5281 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
5341 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
5401 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
5461 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
5521 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
5581 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
5641 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
5701 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
5761 gaag
//
