| 质粒名称: | pAAV.CMV.PI.EGFP.WPRE.bGH |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 105530 |
| 存储实验室: | James M. Wilson |
|
载体骨架基本信息 |
|
| 载体名称: | pAAV |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | -- |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 哺乳动物表达,AAV |
| 筛选标记: | -- |
| 高拷贝/低拷贝: | -- |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | EGFP |
| 物种来源: | -- |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | CMV |
| 标签/融合蛋白: | -- |
| 克隆方法: | -- |
| 5' 克隆位点: | -- |
| 3' 克隆位点: | -- |
| 5' 测序引物 | CMV-F |
| 3' 测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pAAV.CMV.PI.EGFP.WPRE.bGH was a gift from James M. Wilson (Addgene plasmid # 105530 ; http://n2t.net/addgene:105530 ; RRID:Addgene_105530) |
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.CMV.PI.EGFP.WPRE.bGH 5514 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION AAV expression of EGFP from CMV promoter.
ACCESSION .
VERSION .
KEYWORDS pAAV.CMV.PI.EGFP.WPRE.bGH
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5514)
TITLE AAV plasmids
REFERENCE 2 (bases 1 to 5514)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..5514
/organism="synthetic DNA construct"
/mol_type="other DNA"
repeat_region 23..152
/label=AAV2 ITR
enhancer 293..596
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 597..800
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 751..771
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
intron 853..985
/label=chimeric intron
/note="chimera between introns from human beta-globin and
immunoglobulin heavy chain genes"
primer_bind 968..987
/label=pMT2-F
/note="Synthetic intron, forward primer"
regulatory 997..1006
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 1003..1722
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(1048..1069)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(1309..1328)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 1656..1677
/label=EGFP-C
/note="EGFP, forward primer"
primer_bind complement(1789..1809)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(2207..2218)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
primer_bind complement(2285..2302)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 2291..2498
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
repeat_region 2586..2715
/label=AAV2 ITR
primer_bind complement(2734..2751)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(2734..2750)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(2743..2765)
/label=M13/pUC Forward
/note="In lacZ gene"
rep_origin 2892..3347
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2979..2998)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 3189..3210
/label=F1ori-F
/note="F1 origin, forward primer"
promoter 3373..3477
/gene="bla"
/label=AmpR promoter
CDS 3478..4338
/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(3696..3715)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 4509..5097
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 4998..5017
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 5251..5268
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 5385..5406
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 5421..5451
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 5459..5475
/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 5464..5486
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 5483..5499
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 5483..5499
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
ORIGIN
1 tttaattaag gccttaatta ggctgcgcgc tcgctcgctc actgaggccg cccgggcaaa
61 gcccgggcgt cgggcgacct ttggtcgccc ggcctcagtg agcgagcgag cgcgcagaga
121 gggagtggcc aactccatca ctaggggttc cttgtagtta atgattaacc cgccatgcta
181 cttatctacg tagccatgct ctaggaagat cggaattcgc ccttaagcta gctagttatt
241 aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat
301 aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa
361 taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg
421 agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc
481 cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct
541 tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga
601 tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa
661 gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc
721 caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg
781 aggtctatat aagcagagct ggtttagtga accgtcagat cctgcagaag ttggtcgtga
841 ggcactgggc aggtaagtat caaggttaca agacaggttt aaggagacca atagaaactg
901 ggcttgtcga gacagagaag actcttgcgt ttctgatagg cacctattgg tcttactgac
961 atccactttg cctttctctc cacaggtgtc caggcggccg ccatggtgag caagggcgag
1021 gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac
1081 aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag
1141 ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc
1201 tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag
1261 tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac
1321 tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg
1381 aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac
1441 aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc
1501 aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac
1561 acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc
1621 gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc
1681 gccgccggga tcactctcgg catggacgag ctgtacaagt aataagcttg gatccaatca
1741 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt
1801 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc
1861 tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc
1921 cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg
1981 gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc
2041 cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg
2101 cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg
2161 tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc
2221 agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgaga
2281 tctgcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct
2341 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca
2401 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag
2461 ggggaggatt gggaagacaa tagcaggcat gctggggact cgagttaagg gcgaattccc
2521 gataaggatc ttcctagagc atggctacgt agataagtag catggcgggt taatcattaa
2581 ctacaaggaa cccctagtga tggagttggc cactccctct ctgcgcgctc gctcgctcac
2641 tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag
2701 cgagcgagcg cgcagcctta attaacctaa ttcactggcc gtcgttttac aacgtcgtga
2761 ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag
2821 ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa
2881 tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
2941 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
3001 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
3061 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
3121 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
3181 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
3241 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
3301 acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttagg tggcactttt
3361 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
3421 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
3481 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
3541 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
3601 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
3661 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
3721 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
3781 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
3841 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
3901 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
3961 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
4021 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
4081 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
4141 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
4201 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
4261 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
4321 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
4381 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
4441 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
4501 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
4561 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
4621 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
4681 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
4741 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
4801 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
4861 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
4921 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
4981 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
5041 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
5101 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
5161 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
5221 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
5281 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac
5341 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc
5401 actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt
5461 gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc caga
//
