| 质粒名称: | pDS-RED-C1-Beclin1 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 24405 |
| 存储实验室: | Qing Zhong |
|
载体骨架基本信息 |
|
| 载体名称: | pDsRed-C1 |
| 空载体大小: | 4700 bp |
| 完整质粒大小: | 6009 bp |
| 载体类型: | 哺乳细胞表达 |
| 载体抗性: | Kanamycin |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | Beclin1 |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 1350bp |
| 启动子: | CMV |
| 标签/融合蛋白: | DsRed (N terminal on insert) |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | EcoRI(未破坏) |
| 3’克隆位点: | BamHI(未破坏) |
| 5’测序引物 | DsRed1-C |
| 3’测序引物 | SV40pA-R |
|
文献引用方法 |
|
| 材料和方法部分: |
pDS-RED-C1-Beclin1 was a gift from Qing Zhong (Addgene plasmid # 24405 ; http://n2t.net/addgene:24405 ; RRID:Addgene_24405)
|
| 参考文献部分: | Identification of Barkor as a mammalian autophagy-specific factor for Beclin 1 and class III phosphatidylinositol 3-kinase. Sun Q, Fan W, Chen K, Ding X, Chen S, Zhong Q. Proc Natl Acad Sci U S A. 2008 Dec 9. 105(49):19211-6. 10.1073/pnas.0810452105 PubMed 19050071 |
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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 pDS-RED-C1-Beclin1 6009 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pDS-RED-C1-Beclin1
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6009)
AUTHORS Sun Q, Fan W, Chen K, Ding X, Chen S, Zhong Q
TITLE Identification of Barkor as a mammalian autophagy-specific factor
for Beclin 1 and class III phosphatidylinositol 3-kinase.
JOURNAL Proc Natl Acad Sci U S A. 2008 Dec 9. 105(49):19211-6.
PUBMED 19050071
REFERENCE 2 (bases 1 to 6009)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..6009
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 16..319
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 320..523
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 474..494
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
regulatory 562..571
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 568..1242
/codon_start=1
/product="improved tetrameric variant of DsRed fluorescent
protein"
/label=DsRed2
/note="mammalian codon-optimized"
/translation="MASSENVITEFMRFKVRMEGTVNGHEFEIEGEGEGRPYEGHNTVK
LKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
ATVTQDSSLQDGCFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGETHK
ALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDAKLDITSHNEDYTIVEQYERTEGRHH
LFL"
primer_bind complement(748..768)
/label=DsRed1-N
/note="DsRed1, reverse primer"
primer_bind 1160..1183
/label=DsRed1-C
/note="DsRed1, forward primer"
polyA_signal 2752..2873
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(2789..2808)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 2843..2862
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin complement(2880..3335)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(3017..3038)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 3229..3248
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 3362..3466
/gene="bla"
/label=AmpR promoter
promoter 3468..3825
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
primer_bind complement(3491..3511)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 3676..3811
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 3738..3757
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 3860..4654
/codon_start=1
/gene="aph(3')-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/label=NeoR/KanR
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(3914..3933)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 4524..4543
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind complement(4842..4861)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 4886..4933
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 5262..5850
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 5751..5770
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
61 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
121 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
181 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
241 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
301 agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg
361 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg
421 gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat
481 gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctggtttag tgaaccgtca
541 gatccgctag cgctaccggt cgccaccatg gcctcctccg agaacgtcat caccgagttc
601 atgcgcttca aggtgcgcat ggagggcacc gtgaacggcc acgagttcga gatcgagggc
661 gagggcgagg gccgccccta cgagggccac aacaccgtga agctgaaggt gaccaagggc
721 ggccccctgc ccttcgcctg ggacatcctg tccccccagt tccagtacgg ctccaaggtg
781 tacgtgaagc accccgccga catccccgac tacaagaagc tgtccttccc cgagggcttc
841 aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg cgaccgtgac ccaggactcc
901 tccctgcagg acggctgctt catctacaag gtgaagttca tcggcgtgaa cttcccctcc
961 gacggccccg tgatgcagaa gaagaccatg ggctgggagg cctccaccga gcgcctgtac
1021 ccccgcgacg gcgtgctgaa gggcgagacc cacaaggccc tgaagctgaa ggacggcggc
1081 cactacctgg tggagttcaa gtccatctac atggccaaga agcccgtgca gctgcccggc
1141 tactactacg tggacgccaa gctggacatc acctcccaca acgaggacta caccatcgtg
1201 gagcagtacg agcgcaccga gggccgccac cacctgttcc tgagatctcg agctcaagct
1261 tcgaattcta tggaagggtc taagacgtcc aacaacagca ccatgcaggt gagcttcgtg
1321 tgccagcgct gcagccagcc cctgaaactg gacacgagtt tcaagatcct ggaccgtgtc
1381 accatccagg aactcacagc tccattactt accacagccc aggcgaaacc aggagagacc
1441 caggaggaag agactaactc aggagaggag ccatttattg aaactcctcg ccaggatggt
1501 gtctctcgca gattcatccc cccagccagg atgatgtcca cagaaagtgc caacagcttc
1561 actctgattg gggaggcatc tgatggcggc accatggaga acctcagccg aagactgaag
1621 gtcactgggg acctttttga catcatgtcg ggccagacag atgtggatca cccactctgt
1681 gaggaatgca cagatactct tttagaccag ctggacactc agctcaacgt cactgaaaat
1741 gagtgtcaga actacaaacg ctgtttggag atcttagagc aaatgaatga ggatgacagt
1801 gaacagttac agatggagct aaaggagctg gcactagagg aggagaggct gatccaggag
1861 ctggaagacg tggaaaagaa ccgcaagata gtggcagaaa atctcgagaa ggtccaggct
1921 gaggctgaga gactggatca ggaggaagct cagtatcaga gagaatacag tgaatttaaa
1981 cgacagcagc tggagctgga tgatgagctg aagagtgttg aaaaccagat gcgttatgcc
2041 cagacgcagc tggataagct gaagaaaacc aacgtcttta atgcaacctt ccacatctgg
2101 cacagtggac agtttggcac aatcaataac ttcaggctgg gtcgcctgcc cagtgttccc
2161 gtggaatgga atgagattaa tgctgcttgg ggccagactg tgttgctgct ccatgctctg
2221 gccaataaga tgggtctgaa atttcagaga taccgacttg ttccttacgg aaaccattca
2281 tatctggagt ctctgacaga caaatctaag gagctgccgt tatactgttc tggggggttg
2341 cggtttttct gggacaacaa gtttgaccat gcaatggtgg ctttcctgga ctgtgtgcag
2401 cagttcaaag aagaggttga gaaaggcgag acacgttttt gtcttcccta caggatggat
2461 gtggagaaag gcaagattga agacacagga ggcagtggcg gctcctattc catcaaaacc
2521 cagtttaact ctgaggagca gtggacaaaa gctctcaagt tcatgctgac gaatcttaag
2581 tggggtcttg cttgggtgtc ctcacaattt tataacaaat aaggatccac cggatctaga
2641 taactgatca taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc
2701 ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt
2761 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
2821 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttaacgcgta
2881 aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt
2941 ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat
3001 agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa
3061 cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta
3121 atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc
3181 ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc
3241 gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac
3301 acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt
3361 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
3421 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtcctg aggcggaaag
3481 aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
3541 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
3601 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
3661 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
3721 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc
3781 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaagatcg atcaagagac
3841 aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc
3901 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc
3961 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc
4021 cggtgccctg aatgaactgc aagacgaggc agcgcggcta tcgtggctgg ccacgacggg
4081 cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt
4141 gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc
4201 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga
4261 ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga
4321 tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct
4381 caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc
4441 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt
4501 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg
4561 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat
4621 cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc
4681 gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa
4741 aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat
4801 ctcatgctgg agttcttcgc ccaccctagg gggaggctaa ctgaaacacg gaaggagaca
4861 ataccggaag gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacggtgtt
4921 gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca
4981 ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca
5041 agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc
5101 tcaggttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
5161 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
5221 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
5281 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
5341 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
5401 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
5461 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
5521 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
5581 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
5641 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
5701 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
5761 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
5821 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
5881 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
5941 gataaccgta ttaccgccat gcattagtta ttaatagtaa tcaattacgg ggtcattagt
6001 tcatagccc
//
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