| 质粒名称: | pcDNA4-Beclin1 (FL) |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 24388 |
| 存储实验室: | Qing Zhong |
|
载体骨架基本信息 |
|
| 载体名称: | pCDNA4/TO-3*flag |
| 空载体大小: | 5100 bp |
| 完整质粒大小: | 6459 bp |
| 载体类型: | 哺乳细胞表达 |
| 载体抗性: | 氨苄霉素 |
| 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: | 1350 bp |
| 启动子: | CMV |
| 标签/融合蛋白: | 3xFLAG (C terminal on backbone) |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | BamHI (未破坏) |
| 3’克隆位点: | NotI (未破坏) |
| 5’测序引物 | CMV-F |
| 3’测序引物 | BGH-rev |
|
文献引用方法 |
|
| 材料和方法部分: | pcDNA4-Beclin1 (FL) was a gift from Qing Zhong (Addgene plasmid # 24388 ; http://n2t.net/addgene:24388 ; RRID:Addgene_24388) |
| 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 |
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
质粒图谱.png)
载体简介
载体序列
LOCUS pcDNA4-Beclin1 (FL) 6459 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pcDNA4-Beclin1 (FL)
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6459)
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 6459)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..6459
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 143..522
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 523..726
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 677..697
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
protein_bind 728..746
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 749..767
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
primer_bind 777..801
/label=LNCX
/note="Human CMV promoter, forward primer"
CDS 2269..2334
/codon_start=1
/product="three tandem FLAG(R) epitope tags, followed by an
enterokinase cleavage site"
/label=3xFLAG
/translation="DYKDHDGDYKDHDIDYKDDDDK"
primer_bind complement(2375..2392)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 2381..2605
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 2651..3079
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2738..2757)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 2948..2969
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(3088..3108)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 3093..3422
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 3273..3408
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 3335..3354
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
promoter 3470..3517
/label=EM7 promoter
/note="synthetic bacterial promoter "
CDS 3536..3910
/codon_start=1
/gene="Sh ble from Streptoalloteichus hindustanus"
/product="antibiotic-binding protein"
/label=BleoR
/note="confers resistance to bleomycin, phleomycin, and
Zeocin(TM)"
/translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
EFALRDPAGNCVHFVAEEQD"
polyA_signal 4040..4161
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(4077..4096)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 4131..4150
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(4210..4226)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4210..4226)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(4223..4245)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 4234..4250
/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)."
promoter complement(4258..4288)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4303..4324
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(4441..4458)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4612..5200)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4692..4711)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5371..6231)
/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 5994..6013
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(6232..6336)
/gene="bla"
/label=AmpR promoter
primer_bind complement(6411..6430)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
ORIGIN
1 tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc
61 ttgaccgaca attgcatgaa gaatctgctt agggttaggc gttttgcgct gcttcgcgat
121 gtacgggcca gatatacgcg ttgacattga ttattgacta gttattaata gtaatcaatt
181 acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat
241 ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt
301 cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa
361 actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc
421 aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct
481 acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag
541 tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt
601 gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac
661 aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc
721 agagctctcc ctatcagtga tagagatctc cctatcagtg atagagatcg tcgacgagct
781 cgtttagtga accgtcagat cgcctggaga cgccatccac gctgttttga cctccataga
841 agacaccggg accgatccag cctccggact ctagcgttta aacttaagct tggtaccgag
901 ctcggatcca tggaagggtc taagacgtcc aacaacagca ccatgcaggt gagcttcgtg
961 tgccagcgct gcagccagcc cctgaaactg gacacgagtt tcaagatcct ggaccgtgtc
1021 accatccagg aactcacagc tccattactt accacagccc aggcgaaacc aggagagacc
1081 caggaggaag agactaactc aggagaggag ccatttattg aaactcctcg ccaggatggt
1141 gtctctcgca gattcatccc cccagccagg atgatgtcca cagaaagtgc caacagcttc
1201 actctgattg gggaggcatc tgatggcggc accatggaga acctcagccg aagactgaag
1261 gtcactgggg acctttttga catcatgtcg ggccagacag atgtggatca cccactctgt
1321 gaggaatgca cagatactct tttagaccag ctggacactc agctcaacgt cactgaaaat
1381 gagtgtcaga actacaaacg ctgtttggag atcttagagc aaatgaatga ggatgacagt
1441 gaacagttac agatggagct aaaggagctg gcactagagg aggagaggct gatccaggag
1501 ctggaagacg tggaaaagaa ccgcaagata gtggcagaaa atctcgagaa ggtccaggct
1561 gaggctgaga gactggatca ggaggaagct cagtatcaga gagaatacag tgaatttaaa
1621 cgacagcagc tggagctgga tgatgagctg aagagtgttg aaaaccagat gcgttatgcc
1681 cagacgcagc tggataagct gaagaaaacc aacgtcttta atgcaacctt ccacatctgg
1741 cacagtggac agtttggcac aatcaataac ttcaggctgg gtcgcctgcc cagtgttccc
1801 gtggaatgga atgagattaa tgctgcttgg ggccagactg tgttgctgct ccatgctctg
1861 gccaataaga tgggtctgaa atttcagaga taccgacttg ttccttacgg aaaccattca
1921 tatctggagt ctctgacaga caaatctaag gagctgccgt tatactgttc tggggggttg
1981 cggtttttct gggacaacaa gtttgaccat gcaatggtgg ctttcctgga ctgtgtgcag
2041 cagttcaaag aagaggttga gaaaggcgag acacgttttt gtcttcccta caggatggat
2101 gtggagaaag gcaagattga agacacagga ggcagtggcg gctcctattc catcaaaacc
2161 cagtttaact ctgaggagca gtggacaaaa gctctcaagt tcatgctgac gaatcttaag
2221 tggggtcttg cttgggtgtc ctcacaattt tataacaaag gcggccgcga ctacaaagac
2281 catgacggtg attataaaga tcatgacatc gactacaagg atgacgatga caagtgactc
2341 gagtctagag ggcccgttta aacccgctga tcagcctcga ctgtgccttc tagttgccag
2401 ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact
2461 gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt
2521 ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat
2581 gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg
2641 gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
2701 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
2761 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
2821 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
2881 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
2941 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
3001 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
3061 caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc
3121 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
3181 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
3241 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
3301 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
3361 ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
3421 aaaagctccc gggagcttgt atatccattt tcggatctga tcagcacgtg ttgacaatta
3481 atcatcggca tagtatatcg gcatagtata atacgacaag gtgaggaact aaaccatggc
3541 caagttgacc agtgccgttc cggtgctcac cgcgcgcgac gtcgccggag cggtcgagtt
3601 ctggaccgac cggctcgggt tctcccggga cttcgtggag gacgacttcg ccggtgtggt
3661 ccgggacgac gtgaccctgt tcatcagcgc ggtccaggac caggtggtgc cggacaacac
3721 cctggcctgg gtgtgggtgc gcggcctgga cgagctgtac gccgagtggt cggaggtcgt
3781 gtccacgaac ttccgggacg cctccgggcc ggccatgacc gagatcggcg agcagccgtg
3841 ggggcgggag ttcgccctgc gcgacccggc cggcaactgc gtgcacttcg tggccgagga
3901 gcaggactga cacgtgctac gagatttcga ttccaccgcc gccttctatg aaaggttggg
3961 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
4021 ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa
4081 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
4141 caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc
4201 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
4261 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
4321 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
4381 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
4441 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
4501 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
4561 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
4621 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
4681 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
4741 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
4801 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
4861 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
4921 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
4981 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
5041 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
5101 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
5161 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
5221 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
5281 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
5341 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
5401 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
5461 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
5521 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
5581 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
5641 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
5701 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
5761 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
5821 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
5881 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
5941 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
6001 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
6061 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
6121 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
6181 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
6241 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
6301 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
6361 tgacgtcgac ggatcgggag atctcccgat cccctatggt gcactctcag tacaatctgc
6421 tctgatgccg catagttaag ccagtatctg ctccctgct
//
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