| 质粒名称: | pcDNA4-Beclin1-HA |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 24399 |
| 存储实验室: | Qing Zhong |
|
载体骨架基本信息 |
|
| 载体名称: | pCDNA4/TO-HA |
| 空载体大小: | 5100 bp |
| 完整质粒大小: | 6420 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: | 1350bp |
| 标签/融合蛋白: | HA (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-HA was a gift from Qing Zhong (Addgene plasmid # 24399 ; http://n2t.net/addgene:24399 ; RRID:Addgene_24399) |
| 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
载体简介
载体序列
LOCUS pcDNA4-Beclin1-HA 6420 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pcDNA4-Beclin1-HA
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6420)
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 6420)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..6420
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 149..528
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 529..732
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 683..703
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
protein_bind 734..752
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 755..773
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
primer_bind 783..807
/label=LNCX
/note="Human CMV promoter, forward primer"
CDS 2275..2301
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
primer_bind complement(2342..2359)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 2348..2572
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 2618..3046
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2705..2724)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 2915..2936
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(3055..3075)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 3060..3389
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 3240..3375
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 3302..3321
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
promoter 3437..3484
/label=EM7 promoter
/note="synthetic bacterial promoter "
CDS 3503..3877
/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 4007..4128
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(4044..4063)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 4098..4117
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(4177..4193)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4177..4193)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(4190..4212)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 4201..4217
/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(4225..4255)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4270..4291
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(4408..4425)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4579..5167)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4659..4678)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5338..6198)
/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 5961..5980
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(6199..6303)
/gene="bla"
/label=AmpR promoter
primer_bind complement(6378..6397)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
ORIGIN
1 cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg
61 caaggcttga ccgacaattg catgaagaat ctgcttaggg ttaggcgttt tgcgctgctt
121 cgcgatgtac gggccagata tacgcgttga cattgattat tgactagtta ttaatagtaa
181 tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg
241 gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg
301 tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta
361 cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt
421 gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac
481 tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt
541 tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac
601 cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt
661 cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat
721 ataagcagag ctctccctat cagtgataga gatctcccta tcagtgatag agatcgtcga
781 cgagctcgtt tagtgaaccg tcagatcgcc tggagacgcc atccacgctg ttttgacctc
841 catagaagac accgggaccg atccagcctc cggactctag cgtttaaact taagcttggt
901 accgagctcg gatccatgga agggtctaag acgtccaaca acagcaccat gcaggtgagc
961 ttcgtgtgcc agcgctgcag ccagcccctg aaactggaca cgagtttcaa gatcctggac
1021 cgtgtcacca tccaggaact cacagctcca ttacttacca cagcccaggc gaaaccagga
1081 gagacccagg aggaagagac taactcagga gaggagccat ttattgaaac tcctcgccag
1141 gatggtgtct ctcgcagatt catcccccca gccaggatga tgtccacaga aagtgccaac
1201 agcttcactc tgattgggga ggcatctgat ggcggcacca tggagaacct cagccgaaga
1261 ctgaaggtca ctggggacct ttttgacatc atgtcgggcc agacagatgt ggatcaccca
1321 ctctgtgagg aatgcacaga tactctttta gaccagctgg acactcagct caacgtcact
1381 gaaaatgagt gtcagaacta caaacgctgt ttggagatct tagagcaaat gaatgaggat
1441 gacagtgaac agttacagat ggagctaaag gagctggcac tagaggagga gaggctgatc
1501 caggagctgg aagacgtgga aaagaaccgc aagatagtgg cagaaaatct cgagaaggtc
1561 caggctgagg ctgagagact ggatcaggag gaagctcagt atcagagaga atacagtgaa
1621 tttaaacgac agcagctgga gctggatgat gagctgaaga gtgttgaaaa ccagatgcgt
1681 tatgcccaga cgcagctgga taagctgaag aaaaccaacg tctttaatgc aaccttccac
1741 atctggcaca gtggacagtt tggcacaatc aataacttca ggctgggtcg cctgcccagt
1801 gttcccgtgg aatggaatga gattaatgct gcttggggcc agactgtgtt gctgctccat
1861 gctctggcca ataagatggg tctgaaattt cagagatacc gacttgttcc ttacggaaac
1921 cattcatatc tggagtctct gacagacaaa tctaaggagc tgccgttata ctgttctggg
1981 gggttgcggt ttttctggga caacaagttt gaccatgcaa tggtggcttt cctggactgt
2041 gtgcagcagt tcaaagaaga ggttgagaaa ggcgagacac gtttttgtct tccctacagg
2101 atggatgtgg agaaaggcaa gattgaagac acaggaggca gtggcggctc ctattccatc
2161 aaaacccagt ttaactctga ggagcagtgg acaaaagctc tcaagttcat gctgacgaat
2221 cttaagtggg gtcttgcttg ggtgtcctca caattttata acaaaggcgg ccgctaccca
2281 tacgatgttc cagattacgc ttgactcgag tctagagggc ccgtttaaac ccgctgatca
2341 gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc
2401 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg
2461 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg
2521 gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag
2581 gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta
2641 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
2701 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
2761 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
2821 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
2881 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
2941 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
3001 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg
3061 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
3121 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
3181 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
3241 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
3301 ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag
3361 gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg
3421 gatctgatca gcacgtgttg acaattaatc atcggcatag tatatcggca tagtataata
3481 cgacaaggtg aggaactaaa ccatggccaa gttgaccagt gccgttccgg tgctcaccgc
3541 gcgcgacgtc gccggagcgg tcgagttctg gaccgaccgg ctcgggttct cccgggactt
3601 cgtggaggac gacttcgccg gtgtggtccg ggacgacgtg accctgttca tcagcgcggt
3661 ccaggaccag gtggtgccgg acaacaccct ggcctgggtg tgggtgcgcg gcctggacga
3721 gctgtacgcc gagtggtcgg aggtcgtgtc cacgaacttc cgggacgcct ccgggccggc
3781 catgaccgag atcggcgagc agccgtgggg gcgggagttc gccctgcgcg acccggccgg
3841 caactgcgtg cacttcgtgg ccgaggagca ggactgacac gtgctacgag atttcgattc
3901 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
3961 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc
4021 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
4081 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat
4141 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
4201 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
4261 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
4321 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
4381 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
4441 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
4501 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
4561 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
4621 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
4681 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
4741 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
4801 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
4861 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
4921 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
4981 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
5041 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
5101 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
5161 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
5221 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
5281 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
5341 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
5401 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
5461 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
5521 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
5581 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
5641 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
5701 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
5761 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
5821 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
5881 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
5941 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
6001 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
6061 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
6121 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
6181 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
6241 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
6301 gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc tcccgatccc
6361 ctatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gtatctgctc
//
