| 质粒名称: | pscAAV-GFP |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 32396 |
| 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,AAV,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)(未破坏) |
| 3' 克隆位点: | Multiple (see map)(未破坏) |
| 5' 测序引物 | TGGCACCAAAATCAACGGGACTTTCCAAAATGT |
| 3' 测序引物 | AAAACCTCCCACACCTCCCCCTGAAC |
| 文献引用方法 | |
| 材料和方法部分: | pscAAV-GFP was a gift from John T Gray (Addgene plasmid # 32396 ; http://n2t.net/addgene:32396 ; RRID:Addgene_32396) |
| 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
pscAAVGFP载体基本信息
pscAAVGFP载体图谱质粒图谱和多克隆位点信息
pscAAVGFP载体简介
pscAAVGFP载体序列
LOCUS pscAAV-GFP 4369 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pscAAV-GFP
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4369)
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 4369)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..4369
/organism="synthetic DNA construct"
/mol_type="other DNA"
protein_bind 107..128
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 143..173
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 181..197
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
primer_bind 186..208
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 205..221
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 205..221
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
promoter 571..774
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 725..745
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 771..795
/label=LNCX
/note="Human CMV promoter, forward primer"
regulatory 936..945
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 942..1661
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(987..1008)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(1248..1267)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 1595..1616
/label=EGFP-C
/note="EGFP, forward primer"
primer_bind 1705..1724
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind complement(1974..1991)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(1974..1990)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(1983..2005)
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind complement(2199..2218)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 2318..2340
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(2378..2396)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 2464..2568
/gene="bla"
/label=AmpR promoter
CDS 2569..3429
/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(2787..2806)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 3600..4188
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 4089..4108
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 4342..4359
/label=L4440
/note="L4440 vector, forward primer"
ORIGIN
1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagctct
241 cgagatctag aaagcttccc ggggggatct gggccactcc ctctctgcgc gctcgctcgc
301 tcactgaggc cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag
361 tgagcgagcg agcgcgcaga gagggagtgg ccaactccat cactaggggt tcctggaggg
421 gtggagtcgt gacctaggca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa
481 tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac
541 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg
601 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg
661 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca
721 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta
781 gtgaaccgtc agatcgcctg gagacgccat ccggactcta aggtaaatat aaaattttta
841 agtgtataat gtgttaaact actgattcta attgtttctc tcttttagat tccaaccttt
901 ggaactgaat tccgcgggcc cgggatccac cggtcgccac catggtgagc aagggcgagg
961 agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca
1021 agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt
1081 tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct
1141 acggcgtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt
1201 ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact
1261 acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga
1321 agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca
1381 acagccacaa cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca
1441 agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca
1501 cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtccg
1561 ccctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg
1621 ccgccgggat cactctcggc atggacgagc tgtacaagta aagcggccgc taggcctcac
1681 ctgcgatctc gatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt
1741 ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag
1801 ggggaggtgt gggaggtttt ttaaactagt ccactccctc tctgcgcgct cgctcgctca
1861 ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga
1921 gcgagcgagc gcgcagagag ggacagatcc gggcccgcat gcgtcgacaa ttcactggcc
1981 gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca
2041 gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc
2101 caacagttgc gcagcctgaa tggcgaatgg cgcctgatgc ggtattttct ccttacgcat
2161 ctgtgcggta tttcacaccg catatggtgc actctcagta caatctgctc tgatgccgca
2221 tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg
2281 ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg
2341 ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg cctattttta
2401 taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat
2461 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
2521 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
2581 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
2641 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
2701 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
2761 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
2821 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
2881 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
2941 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
3001 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
3061 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
3121 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
3181 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
3241 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
3301 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
3361 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
3421 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
3481 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
3541 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
3601 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
3661 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
3721 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
3781 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
3841 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
3901 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
3961 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
4021 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
4081 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
4141 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
4201 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
4261 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
4321 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaag
//
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