| 质粒名称: | pAAV-GFP |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 32395 |
| 存储实验室: | John T Gray |
|
载体骨架基本信息 |
|
| 载体名称: | pSub201 |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 8310 bp |
| 载体类型: | 哺乳动物表达；腺相关病毒 |
| 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 |
| 启动子 | CMV |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | Multiple (see map) (not destroyed) |
| 3' 克隆位点: | Multiple (see map) (not destroyed) |
| 5' 测序引物 | GGCCCTTTTGCTAATCATGTTCATACCTCTTATCT |
| 3' 测序引物 | GGCCAGGGCATTAGCCACACCAG |
|
文献引用方法 |
|
| 材料和方法部分: | pAAV-GFP was a gift from John T Gray (Addgene plasmid # 32395 ; http://n2t.net/addgene:32395 ; RRID:Addgene_32395) |
| 参考文献部分: | Design and Construction of Functional AAV Vectors. Gray JT, Zolotukhin S. Methods Mol Biol. 2011;807:25-46. 10.1007/978-1-61779-370-7_2 PubMed 22034025 |
买家导航
pAAVGFP载体基本信息
pAAVGFP载体图谱质粒图谱和多克隆位点信息
pAAVGFP载体简介
pAAVGFP载体序列
LOCUS pAAV-GFP 6891 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pAAV-GFP
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6891)
AUTHORS Gray JT, Zolotukhin S
TITLE Design and Construction of Functional AAV Vectors.
JOURNAL Methods Mol Biol. 2011;807:25-46.
PUBMED 22034025
REFERENCE 2 (bases 1 to 6891)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6891
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 85..102
/label=L4440
/note="L4440 vector, forward primer"
repeat_region 168..297
/label=AAV2 ITR
primer_bind complement(353..372)
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
enhancer 474..853
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 854..1057
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 1008..1028
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 1054..1078
/label=LNCX
/note="Human CMV promoter, forward primer"
intron 1171..1646
/label=-beta-globin intron
/note="internally truncated intron from human beta-globin"
regulatory 1724..1733
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 1730..2449
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(1775..1796)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(2036..2055)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 2383..2404
/label=EGFP-C
/note="EGFP, forward primer"
primer_bind complement(2479..2495)
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(2480..2496)
/label=pBluescriptKS
/note="For pBluescript vector"
polyA_site 2589..2983
/label=-beta-globin poly(A)
/note="human beta-globin polyadenylation signal"
repeat_region 3338..3467
/label=AAV2 ITR
rep_origin 3838..4351
/direction=RIGHT
/label=M13 ori
/note="M13 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(3925..3944)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 4135..4156
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(4833..4852)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 4952..4974
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(5012..5030)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 5098..5202
/gene="bla"
/label=AmpR promoter
CDS 5203..6063
/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(5421..5440)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 6234..6822
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 6723..6742
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
61 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca
121 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg cgcgctcgct
181 cgctcactga ggccgcccgg gcaaagcccg ggcgtcgggc gacctttggt cgcccggcct
241 cagtgagcga gcgagcgcgc agagagggag tggccaactc catcactagg ggttccttgt
301 agttaatgat taacccgcca tgctacttat ctacgtagcc atgctctagc ggcctcggcc
361 tctgcataaa taaaaaaaat tagtcagcca tgagcttggc ccattgcata cgttgtatcc
421 atatcataat atgtacattt atattggctc atgtccaaca ttaccgccat gttgacattg
481 attattgact agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat
541 ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc
601 ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca
661 ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta
721 tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta
781 tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat
841 cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga
901 ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca
961 aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg
1021 taggcgtgta cggtgggagg tctatataag cagagctcgt ttagtgaacc gtcagatcgc
1081 ctggagacgc catccacgct gttttgacct ccatagaaga caccgggacc gatccagcct
1141 cccctcgaag ctgatcctga gaacttcagg gtgagtctat gggacccttg atgttttctt
1201 tccccttctt ttctatggtt aagttcatgt cataggaagg ggagaagtaa cagggtacac
1261 atattgacca aatcagggta attttgcatt tgtaatttta aaaaatgctt tcttctttta
1321 atatactttt ttgtttatct tatttctaat actttcccta atctctttct ttcagggcaa
1381 taatgataca atgtatcatg cctctttgca ccattctaaa gaataacagt gataatttct
1441 gggttaaggc aatagcaata tttctgcata taaatatttc tgcatataaa ttgtaactga
1501 tgtaagaggt ttcatattgc taatagcagc tacaatccag ctaccattct gcttttattt
1561 tatggttggg ataaggctgg attattctga gtccaagcta ggcccttttg ctaatcatgt
1621 tcatacctct tatcttcctc ccacagctcc tgggcaacgt gctggtctgt gtgctggccc
1681 atcactttgg caaagaattc cgcgggcccg ggatccaccg gtcgccacca tggtgagcaa
1741 gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa
1801 cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac
1861 cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac
1921 cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc agcacgactt
1981 cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga
2041 cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat
2101 cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta
2161 caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggt
2221 gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca
2281 gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac
2341 ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt
2401 cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtaaa gcggccgctc
2461 tagaggatcc aagcttatcg ataccgtcga cctcgagggc ccagatctaa ttcaccccac
2521 cagtgcaggc tgcctatcag aaagtggtgg ctggtgtggc taatgccctg gcccacaagt
2581 atcactaagc tcgctttctt gctgtccaat ttctattaaa ggttcctttg ttccctaagt
2641 ccaactacta aactggggga tattatgaag ggccttgagc atctggattc tgcctaataa
2701 aaaacattta ttttcattgc aatgatgtat ttaaattatt tctgaatatt ttactaaaaa
2761 gggaatgtgg gaggtcagtg catttaaaac ataaagaaat gaagagctag ttcaaacctt
2821 gggaaaatac actatatctt aaactccatg aaagaaggtg aggctgcaaa cagctaatgc
2881 acattggcaa cagcccctga tgcctatgcc ttattcatcc ctcagaaaag gattcaagta
2941 gaggcttgat ttggaggtta aagttttgct atgctgtatt ttacattact tattgtttta
3001 gctgtcctca tgaatgtctt ttcactaccc atttgcttat cctgcatctc tcagccttga
3061 ctccactcag ttctcttgct tagagatacc acctttcccc tgaagtgttc cttccatgtt
3121 ttacggcgag atggtttctc ctcgcctggc cactcagcct tagttgtctc tgttgtctta
3181 tagaggtcta cttgaagaag gaaaaacagg gggcatggtt tgactgtcct gtgagccctt
3241 cttccctgcc tcccccactc acagtgaccc ggaatccctc gacatctaga gcatggctac
3301 gtagataagt agcatggcgg gttaatcatt aactacaagg aacccctagt gatggagttg
3361 gccactccct ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga
3421 cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg gcgtaatagc
3481 gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggcga
3541 ttccgttgca atggctggcg gtaatattgt tctggatatt accagcaagg ccgatagttt
3601 gagttcttct actcaggcaa gtgatgttat tactaatcaa agaagtattg cgacaacggt
3661 taatttgcgt gatggacaga ctcttttact cggtggcctc actgattata aaaacacttc
3721 tcaggattct ggcgtaccgt tcctgtctaa aatcccttta atcggcctcc tgtttagctc
3781 ccgctctgat tctaacgagg aaagcacgtt atacgtgctc gtcaaagcaa ccatagtacg
3841 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
3901 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
3961 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
4021 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
4081 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
4141 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
4201 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
4261 cgaattttaa caaaatatta acgcttacaa tttaaatatt tgcttataca atcttcctgt
4321 ttttggggct tttctgatta tcaaccgggg tacatatgat tgacatgcta gttttacgat
4381 taccgttcat cgattctctt gtttgctcca gactctcagg caatgacctg atagcctttg
4441 tagagacctc tcaaaaatag ctaccctctc cggcatgaat ttatcagcta gaacggttga
4501 atatcatatt gatggtgatt tgactgtctc cggcctttct cacccgtttg aatctttacc
4561 tacacattac tcaggcattg catttaaaat atatgagggt tctaaaaatt tttatccttg
4621 cgttgaaata aaggcttctc ccgcaaaagt attacagggt cataatgttt ttggtacaac
4681 cgatttagct ttatgctctg aggctttatt gcttaatttt gctaattctt tgccttgcct
4741 gtatgattta ttggatgttg gaatcgcctg atgcggtatt ttctccttac gcatctgtgc
4801 ggtatttcac accgcatatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
4861 agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg
4921 gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca
4981 ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt
5041 aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc
5101 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
5161 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
5221 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
5281 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
5341 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
5401 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
5461 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
5521 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
5581 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
5641 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
5701 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
5761 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
5821 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
5881 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
5941 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
6001 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
6061 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
6121 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
6181 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
6241 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
6301 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
6361 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
6421 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
6481 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
6541 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
6601 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
6661 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
6721 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
6781 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
6841 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc t
//
