| 质粒名称: | FLAG.PKCepsilon |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 10795 |
| 存储实验室: | Alex Toker |
|
载体骨架基本信息 |
|
| 载体名称: | pcDNA3 |
| 载体出品公司: | Invitrogen |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 5446 bp |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | 新霉素（G418） |
| 高拷贝/低拷贝: | 高拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | PKC epsilon |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 2100 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | FLAG（插入N端子） |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | BamHI(未破坏) |
| 3' 克隆位点: | XbaI (未破坏) |
| 5' 测序引物 | T7 |
| 3' 测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | FLAG.PKCepsilon was a gift from Alex Toker (Addgene plasmid # 10795 ; http://n2t.net/addgene:10795 ; RRID:Addgene_10795) |
| 参考文献部分: | Regulation of novel protein kinase C epsilon by phosphorylation. Cenni V, Doppler H, Sonnenburg ED, Maraldi N, Newton AC, Toker A. Biochem J. 2002 May 1. 363(Pt 3):537-45. 10.1042/0264-6021:3630537 PubMed 11964154 |
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 FLAG.PKCepsilon 7616 bp ds-DNA circular SYN 26-MAR-2019
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS FLAG.PKCepsilon
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7616)
AUTHORS Cenni V, Doppler H, Sonnenburg ED, Maraldi N, Newton AC, Toker A
TITLE Regulation of novel protein kinase C epsilon by phosphorylation.
JOURNAL Biochem J. 2002 May 1. 363(Pt 3):537-45.
PUBMED 11964154
REFERENCE 2 (bases 1 to 7616)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Mar 26, 2019 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..7616
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(42..61)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 233..612
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 613..816
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 767..787
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 861..880
/label=T7
/note="T7 promoter, forward primer"
promoter 861..879
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 917..940
/codon_start=1
/product="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/label=FLAG
/translation="DYKDDDDK"
CDS 1585..1596
/codon_start=1
/gene="cspA"
/product="translation enhancing element for E. coli (Qing
et al., 2004)"
/label=TEE
/translation="NHKV"
promoter complement(3167..3185)
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind complement(3168..3185)
/label=SP6
/note="SP6 promoter, forward primer"
primer_bind complement(3205..3222)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 3211..3435
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 3481..3909
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(3568..3587)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 3778..3799
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(3918..3938)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 3923..4252
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 4103..4238
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 4165..4184
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 4319..5113
/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
GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(4373..4392)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 4983..5002
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
polyA_signal 5287..5408
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(5324..5343)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 5378..5397
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(5457..5473)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(5457..5473)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(5470..5492)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 5481..5497
/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(5505..5535)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 5550..5571
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(5688..5705)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(5859..6447)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(5939..5958)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(6618..7478)
/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 7241..7260
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(7479..7583)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 cggatcggga gatctcccga tcccctatgg tgcactctca gtacaatctg ctctgatgcc
61 gcatagttaa gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg
121 agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt
181 agggttaggc gttttgcgct gcttcgcgat gtacgggcca gatatacgcg ttgacattga
241 ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg
301 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
361 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat
421 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
481 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
541 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc
601 gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac
661 tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa
721 aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt
781 aggcgtgtac ggtgggaggt ctatataagc agagctctct ggctaactag agaacccact
841 gcttactggc ttatcgaaat taatacgact cactataggg agacccaagc ttggtaccga
901 gctcggatcc accatggact ataaggacga tgatgacaaa gtagtgttca atggccttct
961 taagatcaaa atctgcgagg ccgtgagctt gaagcccaca gcctggtcgc tgcgccatgc
1021 ggtgggaccc cggccgcaca ctttccttct cgacccctac attgccctca atgtggacga
1081 ctcgcgcatc ggccaaacgg ccaccaagca gaagaccaac agcccggcct ggcacgacga
1141 gttcgtcacc gatgtgtgca acggacgcaa gatcgagctg gctgtctttc acgatgcccc
1201 cataggctac gacgacttcg tggccaactg caccatccag tttgaggagc tgctgcagaa
1261 cgggagccgc cacttcgagg actggattga tctggagcca gaaggaagag tgtatgtgat
1321 catcgatctc tcagggtcgt cgggtgaagc ccctaaagac aatgaagagc gtgtgttcag
1381 ggaacgcatg cggccgagga agcggcaggg ggccgtcagg cgcagggtcc atcaggtcaa
1441 cggccacaag ttcatggcca cctatcttcg gcagcccacc tactgctccc attgcagaga
1501 cttcatctgg ggtgtcatag gaaagcaggg ataccagtgt caagtctgca cctgcgtggt
1561 ccacaagcgg tgccacgagc tcataatcac aaagtgtgct gggttaaaga agcaggagac
1621 ccccgaccag gtgggctccc agcggttcag cgtcaacatg ccccacaagt tcggtatcca
1681 caactacaag gtccctacct tctgcgatca ctgtgggtcc ctgctctggg gactcttgcg
1741 gcagggtttg cagtgtaaag tctgcaaaat gaatgttcac cgtcgatgtg agaccaacgt
1801 ggctcccaac tgtggagtgg atgccagagg aatcgccaaa gtactggccg acctgggcgt
1861 taccccagac aaaatcacca acagcggcca gagaaggaaa aagctcattg ctggtgccga
1921 gtccccgcag cctgcttctg gaagctcacc atctgaggaa gatcgatcca agtcagcacc
1981 cacctcccct tgtgaccagg aaataaaaga acttgagaac aacattcgga aagccttgtc
2041 atttgacaac cgaggagagg agcaccgggc agcatcgtct cctgatggcc agctgatgag
2101 ccccggtgag aatggcgaag tccggcaagg ccaggccaag cgcctgggcc tggatgagtt
2161 caacttcatc aaggtgttgg gcaaaggcag ctttggcaag gtcatgttgg cagaactcaa
2221 gggcaaagat gaagtatatg ctgtgaaggt cttaaagaag gacgtcatcc ttcaggatga
2281 tgacgtggac tgcacaatga cagagaagag gattttggct ctggcacgga aacacccgta
2341 ccttacccaa ctctactgct gcttccagac caaggaccgc ctctttttcg tcatggaata
2401 tgtaaatggt ggagacctca tgtttcagat tcagcgctcc cgaaaattcg acgagcctcg
2461 ttcacggttc tatgctgcag aggtcacatc ggccctcatg ttcctccacc agcatggagt
2521 catctacagg gatttgaaac tggacaacat ccttctggat gcagaaggtc actgcaagct
2581 ggctgacttc gggatgtgca aggaagggat tctgaatggt gtgacgacca ccacgttctg
2641 tgggactcct gactacatag ctcctgagat cctgcaggag ttggagtatg gcccctccgt
2701 ggactggtgg gccctggggg tgctgatgta cgagatgatg gctggacagc ctccctttga
2761 ggccgacaat gaggacgacc tatttgagtc catcctccat gacgacgtgc tgtacccagt
2821 ctggctcagc aaggaggctg tcagcatctt gaaagctttc atgacgaaga atccccacaa
2881 gcgcctgggc tgtgtggcat cgcagaatgg cgaggacgcc atcaagcagc acccattctt
2941 caaagagatt gactgggtgc tcctggagca gaagaagatc aagccaccct tcaaaccacg
3001 cattaaaacc aaaagagacg tcaataattt tgaccaagac tttacccggg aagagccggt
3061 actcaccctt gtggacgaag caattgtaaa gcagatcaac caggaggaat tcaaaggttt
3121 ctcctacttt ggtgaagacc tgatgccctg atctagaggg ccctattcta tagtgtcacc
3181 taaatgctag agctcgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg
3241 tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct
3301 aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg
3361 gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg
3421 cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc
3481 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
3541 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
3601 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
3661 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
3721 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
3781 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
3841 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
3901 acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
3961 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
4021 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
4081 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
4141 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga
4201 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc
4261 gggagcttgt atatccattt tcggatctga tcaagagaca ggatgaggat cgtttcgcat
4321 gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg
4381 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc
4441 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca
4501 ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct
4561 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga
4621 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg
4681 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat
4741 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga
4801 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg
4861 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg
4921 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat
4981 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct
5041 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga
5101 cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg
5161 ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt
5221 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc
5281 caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
5341 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
5401 gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca
5461 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
5521 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
5581 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
5641 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
5701 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
5761 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
5821 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
5881 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
5941 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
6001 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
6061 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
6121 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
6181 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
6241 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
6301 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
6361 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
6421 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
6481 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
6541 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
6601 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
6661 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
6721 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
6781 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
6841 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
6901 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
6961 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
7021 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
7081 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
7141 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
7201 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
7261 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
7321 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
7381 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
7441 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
7501 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
7561 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcga
//
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