| 质粒名称: | pCDNA3 Flag p38 alpha |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 20351 |
| 存储实验室: | Roger Davis |
|
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: | 5500 bp |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 哺乳动物的表达 |
| 筛选标记: | -- |
| 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: | pDCNA3 Flag p38 alpha,MapK14 |
| 物种来源: | M. musculus (mouse) |
| 插入大小: | 1127 bp |
| 基因突变: | -- |
| GenBank编号 | NM_011951 |
| 启动子 | -- |
| 标签/融合蛋白: | Flag（插入时N端子） |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | Hind III(未破坏) |
| 3' 克隆位点: | Xba I (未破坏) |
| 5' 测序引物 | T7 |
| 3' 测序引物 | SP6 |
|
文献引用方法 |
|
| 材料和方法部分: | pCDNA3 Flag p38 alpha was a gift from Roger Davis (Addgene plasmid # 20351 ; http://n2t.net/addgene:20351 ; RRID:Addgene_20351) |
| 参考文献部分: | Selective activation of p38 mitogen-activated protein (MAP) kinase isoforms by the MAP kinase kinases MKK3 and MKK6. Enslen H, Raingeaud J, Davis RJ. J Biol Chem. 1998 Jan 16. 273(3):1741-8. 10.1074/jbc.273.3.1741 PubMed 9430721 |
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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 pCDNA3 Flag p38 alpha 6486 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pCDNA3 Flag p38 alpha
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6486)
AUTHORS Enslen H, Raingeaud J, Davis RJ
TITLE Selective activation of p38 mitogen-activated protein (MAP) kinase
isoforms by the MAP kinase kinases MKK3 and MKK6.
JOURNAL J Biol Chem. 1998 Jan 16. 273(3):1741-8.
PUBMED 9430721
REFERENCE 2 (bases 1 to 6486)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6486
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(63..80)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(234..822)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(314..333)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(993..1853)
/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 1616..1635
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(1854..1958)
/gene="bla"
/label=AmpR promoter
primer_bind complement(2033..2052)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 2224..2603
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 2604..2807
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 2758..2778
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 2852..2871
/label=T7
/note="T7 promoter, forward primer"
promoter 2852..2870
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 2898..2921
/codon_start=1
/product="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/label=FLAG
/translation="DYKDDDDK"
promoter complement(4028..4046)
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind complement(4029..4046)
/label=SP6
/note="SP6 promoter, forward primer"
primer_bind complement(4066..4083)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 4072..4296
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 4342..4770
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(4429..4448)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 4639..4660
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(4779..4799)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 4784..5113
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 4964..5099
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 5026..5045
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 5180..5974
/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(5234..5253)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 5844..5863
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
polyA_signal 6148..6269
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(6185..6204)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 6239..6258
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(6318..6334)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(6318..6334)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(6331..6353)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 6342..6358
/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(6366..6396)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 6411..6432
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
ORIGIN
1 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
61 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
121 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
181 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
241 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
301 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
361 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
421 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
481 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
541 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
601 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
661 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
721 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
781 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
841 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
901 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
961 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
1021 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
1081 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
1141 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
1201 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
1261 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
1321 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
1381 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
1441 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
1501 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
1561 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
1621 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
1681 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
1741 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
1801 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
1861 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
1921 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
1981 cctgacgtcg acggatcggg agatctcccg atcccctatg gtgcactctc agtacaatct
2041 gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg
2101 agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga
2161 agaatctgct tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc
2221 gttgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
2281 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
2341 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
2401 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
2461 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
2521 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
2581 tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat
2641 agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt
2701 tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc
2761 aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctctc tggctaacta
2821 gagaacccac tgcttactgg cttatcgaaa ttaatacgac tcactatagg gagacccaag
2881 cttcggcagc caacatggac tataaggacg atgatgacaa aatgtcgcag gagaggccca
2941 cgttctaccg gcaggagctg aacaagacca tctgggaggt gcccgaacga taccagaacc
3001 tgtccccggt gggctcgggc gcctatggct cggtgtgtgc tgcttttgat acaaagacgg
3061 ggcatcgtgt ggcagttaag aagctgtcga gaccgtttca gtccatcatt cacgccaaaa
3121 ggacctaccg agagttgcgt ctgctgaagc acatgaaaca cgaaaatgtg attggtctgt
3181 tggatgtgtt cacacccgca aggtcactgg aggaattcaa tgacgtgtac ctggtgaccc
3241 atctcatggg ggcggacctg aacaacatcg tgaagtgcca gaagctgacc gacgaccacg
3301 ttcagtttct catctaccag atcctccgag ggctgaagta tatacattcg gctgacataa
3361 ttcacaggga cctaaagccc agcaacctag ctgtgaacga agactgtgag ctcaagattc
3421 tggattttgg gctggctcgg cacactgatg atgagatgac aggctacgtg gctaccaggt
3481 ggtaccgagc cccagagatc atgctgaatt ggatgcacta taaccagaca gtggatattt
3541 ggtccgtggg ctgcatcatg gctgagctgt tgaccggaag aacgttgttt cctggtacag
3601 accatattga tcagttgaag ctcattttaa gactcgttgg aaccccaggg gctgagcttc
3661 tgaagaaaat ctcctcagag tctgcaagaa actacattca gtctctggcc cagatgccga
3721 agatgaactt cgcaaatgta tttattggtg ccaatcccct ggctgtcgac ctactggaga
3781 agatgctcgt tttggactca gataagagga tcacagcagc ccaagctctt gcgcatgcct
3841 actttgctca gtaccacgac cctgatgatg agcctgttgc tgacccttat gaccagtcct
3901 ttgaaagcag ggaccttctc atagatgagt ggaagagcct gacctatgat gaagtcatca
3961 gctttgtgcc accacccctt gaccaagaag aaatggagtc ctgagcacct ggtctagagg
4021 gccctattct atagtgtcac ctaaatgcta gagctcgctg atcagcctcg actgtgcctt
4081 ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg
4141 ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt
4201 gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca
4261 atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct
4321 ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
4381 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
4441 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
4501 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
4561 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
4621 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
4681 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
4741 agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg
4801 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
4861 agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
4921 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
4981 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
5041 cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
5101 aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcaagagac
5161 aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc
5221 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc
5281 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc
5341 cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg
5401 cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt
5461 gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc
5521 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga
5581 ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga
5641 tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct
5701 caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc
5761 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt
5821 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg
5881 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat
5941 cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc
6001 gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa
6061 aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat
6121 ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa
6181 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
6241 ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag
6301 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
6361 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
6421 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
6481 cagctg
//
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