| 质粒名称: | pcDNA4-Beclin1 151-241 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 24390 |
| 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: | 5100bp |
| 完整质粒大小: | 5388 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: | 270bp |
| 基因突变: | Contains amino acids 151-241. |
| 标签/融合蛋白: |
FLAG (C terminal on insert)
FLAG (C terminal on insert)
FLAG (C terminal on insert)
|
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | BamHI (unknown if destroyed) |
| 3’克隆位点: | NotI (unknown if destroyed) |
| 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 151-241 was a gift from Qing Zhong (Addgene plasmid # 24390 ; http://n2t.net/addgene:24390 ; RRID:Addgene_24390) |
| 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 pcDNA4-Beclin1 151-241 5388 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pcDNA4-Beclin1 151-241
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5388)
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 5388)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..5388
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 1..380
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 381..584
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 535..555
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
protein_bind 586..604
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 607..625
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
primer_bind 635..659
/label=LNCX
/note="Human CMV promoter, forward primer"
CDS 1056..1121
/codon_start=1
/product="three tandem FLAG(R) epitope tags, followed by an
enterokinase cleavage site"
/label=3xFLAG
/translation="DYKDHDGDYKDHDIDYKDDDDK"
primer_bind complement(1162..1179)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 1168..1392
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 1438..1866
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(1525..1544)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 1735..1756
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(1875..1895)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 1880..2209
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 2060..2195
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 2122..2141
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
promoter 2257..2304
/label=EM7 promoter
/note="synthetic bacterial promoter "
CDS 2323..2697
/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 2827..2948
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(2864..2883)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 2918..2937
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(2997..3013)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(2997..3013)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(3010..3032)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 3021..3037
/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(3045..3075)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 3090..3111
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(3228..3245)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(3399..3987)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(3479..3498)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(4158..5018)
/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 4781..4800
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(5019..5123)
/gene="bla"
/label=AmpR promoter
primer_bind complement(5198..5217)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
ORIGIN
1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctccct atcagtgata
601 gagatctccc tatcagtgat agagatcgtc gacgagctcg tttagtgaac cgtcagatcg
661 cctggagacg ccatccacgc tgttttgacc tccatagaag acaccgggac cgatccagcc
721 tccggactct agcgtttaaa cttaagcttg gtaccgagct cggatccatg cagctcaacg
781 tcactgaaaa tgagtgtcag aactacaaac gctgtttgga gatcttagag caaatgaatg
841 aggatgacag tgaacagtta cagatggagc taaaggagct ggcactagag gaggagaggc
901 tgatccagga gctggaagac gtggaaaaga accgcaagat agtggcagaa aatctcgaga
961 aggtccaggc tgaggctgag agactggatc aggaggaagc tcagtatcag agagaataca
1021 gtgaatttaa acgacagcag ctggagggcg gccgcgacta caaagaccat gacggtgatt
1081 ataaagatca tgacatcgac tacaaggatg acgatgacaa gtgactcgag tctagagggc
1141 ccgtttaaac ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt
1201 gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat
1261 aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg
1321 tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg
1381 tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg
1441 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
1501 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
1561 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
1621 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
1681 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
1741 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
1801 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
1861 cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
1921 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
1981 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
2041 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
2101 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct
2161 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg
2221 agcttgtata tccattttcg gatctgatca gcacgtgttg acaattaatc atcggcatag
2281 tatatcggca tagtataata cgacaaggtg aggaactaaa ccatggccaa gttgaccagt
2341 gccgttccgg tgctcaccgc gcgcgacgtc gccggagcgg tcgagttctg gaccgaccgg
2401 ctcgggttct cccgggactt cgtggaggac gacttcgccg gtgtggtccg ggacgacgtg
2461 accctgttca tcagcgcggt ccaggaccag gtggtgccgg acaacaccct ggcctgggtg
2521 tgggtgcgcg gcctggacga gctgtacgcc gagtggtcgg aggtcgtgtc cacgaacttc
2581 cgggacgcct ccgggccggc catgaccgag atcggcgagc agccgtgggg gcgggagttc
2641 gccctgcgcg acccggccgg caactgcgtg cacttcgtgg ccgaggagca ggactgacac
2701 gtgctacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt
2761 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc
2821 caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
2881 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
2941 gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca
3001 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
3061 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
3121 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
3181 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
3241 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
3301 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
3361 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
3421 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
3481 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
3541 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
3601 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
3661 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
3721 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
3781 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
3841 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
3901 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
3961 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
4021 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
4081 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
4141 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
4201 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
4261 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
4321 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
4381 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
4441 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
4501 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
4561 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
4621 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
4681 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
4741 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
4801 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
4861 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
4921 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
4981 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
5041 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
5101 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga
5161 tcgggagatc tcccgatccc ctatggtgca ctctcagtac aatctgctct gatgccgcat
5221 agttaagcca gtatctgctc cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca
5281 aaatttaagc tacaacaagg caaggcttga ccgacaattg catgaagaat ctgcttaggg
5341 ttaggcgttt tgcgctgctt cgcgatgtac gggccagata tacgcgtt
//
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