| 质粒名称: | pcDNA4-Vps34-Flag |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 24398 |
| 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: | 7822bp |
| 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: | Vps34 |
| 物种来源: | H. sapiens (human) |
| 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: | NotI(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-Vps34-Flag was a gift from Qing Zhong (Addgene plasmid # 24398 ; http://n2t.net/addgene:24398 ; RRID:Addgene_24398) |
| 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-Vps34-Flag 7822 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pcDNA4-Vps34-Flag
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7822)
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 7822)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..7822
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(44..63)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 235..614
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 615..818
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 769..789
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
protein_bind 820..838
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 841..859
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
primer_bind 869..893
/label=LNCX
/note="Human CMV promoter, forward primer"
regulatory 1048..1057
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 3724..3789
/codon_start=1
/product="three tandem FLAG(R) epitope tags, followed by an
enterokinase cleavage site"
/label=3xFLAG
/translation="DYKDHDGDYKDHDIDYKDDDDK"
primer_bind complement(3830..3847)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 3836..4060
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 4106..4534
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(4193..4212)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 4403..4424
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(4543..4563)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 4548..4877
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 4728..4863
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 4790..4809
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
promoter 4925..4972
/label=EM7 promoter
/note="synthetic bacterial promoter "
CDS 4991..5365
/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 5495..5616
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(5532..5551)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 5586..5605
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(5665..5681)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(5665..5681)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(5678..5700)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 5689..5705
/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(5713..5743)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 5758..5779
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(5896..5913)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(6067..6655)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(6147..6166)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(6826..7686)
/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 7449..7468
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(7687..7791)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ccctatcagt gatagagatc
841 tccctatcag tgatagagat cgtcgacgag ctcgtttagt gaaccgtcag atcgcctgga
901 gacgccatcc acgctgtttt gacctccata gaagacaccg ggaccgatcc agcctccgga
961 ctctagcgtt taaacttaag cttggtaccg agctcggatc cactagtcca gtgtggtgga
1021 attctgcaga tatccagcac agtggcggcc gccatggggg aagcagagaa gtttcactac
1081 atctatagtt gtgacctgga tatcaacgtc cagcttaaga taggaagctt ggaagggaag
1141 agagaacaaa agagttataa agctgtcctg gaagacccaa tgttgaagtt ctcaggacta
1201 tatcaagaga catgctctga tctttatgtt acttgtcaag tttttgcaga agggaagcct
1261 ttggccttgc cagtgagaac atcctacaaa gcatttagta caagatggaa ctggaatgaa
1321 tggctgaaac taccagtaaa ataccctgac ctgcccagga atgcccaagt ggccctcacc
1381 atatgggatg tgtatggtcc cggaaaagca gtgcctgtag gaggaacaac ggtttcgctc
1441 tttggaaaat acggcatgtt tcgccaaggg atgcatgact tgaaagtctg gcctaatgta
1501 gaagcagatg gatcagaacc cacaaaaact cctggcagaa caagtagcac tctctcagaa
1561 gatcagatga gccgtcttgc caagctcacc aaagctcatc gacaaggaca catggtgaaa
1621 gtagattggc tggatagatt gacatttaga gaaatagaaa tgataaatga gagtgaaaaa
1681 cgaagttcta atttcatgta cctgatggtt gaatttcgat gtgtcaagtg tgatgataag
1741 gaatatggta ttgtttatta tgaaaaggac ggtgatgaat catctccaat tttaacaagt
1801 tttgaattag tgaaagttcc tgacccccag atgtctatgg agaatttagt tgagagcaaa
1861 caccacaagc ttgcccggag tttaagaagt ggaccttctg accacgatct gaaacccaat
1921 gctgccacga gagatcagtt aaatattatt gtgagttatc caccaaccaa gcaacttaca
1981 tatgaagaac aagatcttgt ttggaagttt agatattatc ttacgaatca agaaaaagcc
2041 ttgacaaaat tcttgaaatg tgttaattgg gatctacctc aagaggccaa acaggccttg
2101 gaacttctgg gaaaatggaa gccgatggat gtagaggact ccttggagct gttatcctct
2161 cattacacca acccaactgt gaggcgttat gctgttgccc ggttgcgaca ggccgatgat
2221 gaggatttgt tgatgtacct attacaattg gtccaggctc tcaaatatga aaattttgat
2281 gatataaaga atggattgga acctaccaag aaggatagtc agagttcagt gtcagaaaat
2341 gtgtcaaatt ctggaataaa ttctgcagaa atagatagct cccaaattat aaccagcccc
2401 cttccttcag tctcttcacc tcctcctgca tcaaaaacaa aagaagttcc agatggcgaa
2461 aatctggaac aagatctctg taccttcttg atatcgagag cctgcaaaaa ctcaacactg
2521 gctaattatt tatactggta tgtgatagtg gaatgtgaag atcaagatac tcagcagaga
2581 gatccaaaga cccatgagat gtacttgaac gtaatgagaa gattcagcca agcattgttg
2641 aagggtgata agtctgtcag agttatgcgt tctttgctgg ctgcacaaca gacatttgta
2701 gatcggttgg tgcatctaat gaaggcagta caacgcgaaa gtggaaatcg taagaaaaag
2761 aatgagagac tacaggcatt gcttggagat aatgaaaaga tgaatttgtc agatgtggaa
2821 cttatcccgt tgcctttaga accccaagtg aaaattagag gaataattcc ggaaacagct
2881 acactgttta aaagtgccct tatgcctgca cagttgtttt ttaagacgga agatggaggc
2941 aaatatccag ttatatttaa gcatggagat gatttacgtc aagatcaact tattcttcaa
3001 atcatttcac tcatggacaa gctgttacgg aaagaaaatc tggacttgaa attgacacct
3061 tataaggtgt tagccaccag tacaaaacat ggcttcatgc agtttatcca gtcagttcct
3121 gtggctgaag ttcttgatac agagggaagc attcagaact tttttagaaa atatgcacca
3181 agtgagaatg ggccaaatgg gattagtgct gaggtcatgg acacttacgt taaaagctgt
3241 gctggatatt gcgtgatcac ctatatactt ggagttggag acaggcacct ggataacctt
3301 ttgctaacaa aaacaggcaa actcttccac atagactttg gatatatttt gggtcgggat
3361 ccaaagcctc ttcctccacc aatgaagctg aataaagaaa tggtagaagg aatggggggc
3421 acacagagtg agcagtacca agagttccgt aaacagtgtt acacggcttt cctccacctg
3481 cgaaggtatt ctaatctgat tttgaacttg ttttccttga tggttgatgc aaacattcca
3541 gatattgcac ttgaaccaga taaaactgtg aaaaaggttc aggataaatt ccgcttagac
3601 ctgtcggatg aagaggctgt gcattacatg cagagtctga ttgatgagag tgtccatgct
3661 ctttttgctg cagtggtgga acagattcac aagtttgccc agtactggag aaaaggcggc
3721 cgcgactaca aagaccatga cggtgattat aaagatcatg acatcgacta caaggatgac
3781 gatgacaagt gactcgagtc tagagggccc gtttaaaccc gctgatcagc ctcgactgtg
3841 ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa
3901 ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt
3961 aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa
4021 gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc
4081 agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt
4141 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
4201 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
4261 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
4321 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
4381 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
4441 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
4501 aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag
4561 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
4621 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
4681 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
4741 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
4801 aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag
4861 gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcagc
4921 acgtgttgac aattaatcat cggcatagta tatcggcata gtataatacg acaaggtgag
4981 gaactaaacc atggccaagt tgaccagtgc cgttccggtg ctcaccgcgc gcgacgtcgc
5041 cggagcggtc gagttctgga ccgaccggct cgggttctcc cgggacttcg tggaggacga
5101 cttcgccggt gtggtccggg acgacgtgac cctgttcatc agcgcggtcc aggaccaggt
5161 ggtgccggac aacaccctgg cctgggtgtg ggtgcgcggc ctggacgagc tgtacgccga
5221 gtggtcggag gtcgtgtcca cgaacttccg ggacgcctcc gggccggcca tgaccgagat
5281 cggcgagcag ccgtgggggc gggagttcgc cctgcgcgac ccggccggca actgcgtgca
5341 cttcgtggcc gaggagcagg actgacacgt gctacgagat ttcgattcca ccgccgcctt
5401 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
5461 cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg
5521 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
5581 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc
5641 tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
5701 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
5761 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
5821 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
5881 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
5941 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
6001 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
6061 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
6121 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
6181 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
6241 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
6301 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
6361 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
6421 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
6481 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
6541 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
6601 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
6661 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
6721 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
6781 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
6841 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
6901 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
6961 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
7021 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
7081 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
7141 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
7201 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
7261 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
7321 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
7381 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
7441 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
7501 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
7561 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
7621 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
7681 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
7741 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
7801 ccgaaaagtg ccacctgacg tc
//
