| 质粒名称: | pCDNA3-Flag MKK6 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 13517 |
| 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: | 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: | MKK6 |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 1248 bp |
| 基因突变: | -- |
| GenBank编号 | U39656 |
| 启动子 | CMV |
| 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: | Hind III (未破坏) |
| 3' 克隆位点: | Xba I(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: | pCDNA3-Flag MKK6 was a gift from Roger Davis (Addgene plasmid # 13517 ; http://n2t.net/addgene:13517 ; RRID:Addgene_13517) |
| 参考文献部分: | MKK3- and MKK6-regulated gene expression is mediated by the p38 mitogen-activated protein kinase signal transduction pathway. Raingeaud J, Whitmarsh AJ, Barrett T, Derijard B, Davis RJ. Mol Cell Biol. 1996 Mar . 16(3):1247-55. PubMed 8622669 |
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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 MKK6 6589 bp ds-DNA circular SYN 19-SEP-2018
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pCDNA3-Flag MKK6
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6589)
AUTHORS Raingeaud J, Whitmarsh AJ, Barrett T, Derijard B, Davis RJ
TITLE MKK3- and MKK6-regulated gene expression is mediated by the p38
mitogen-activated protein kinase signal transduction pathway.
JOURNAL Mol Cell Biol. 1996 Mar . 16(3):1247-55.
PUBMED 8622669
REFERENCE 2 (bases 1 to 6589)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Sep 19, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6589
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(46..65)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 237..616
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 617..820
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 771..791
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 865..884
/label=T7
/note="T7 promoter, forward primer"
promoter 865..883
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 911..934
/codon_start=1
/product="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/label=FLAG
/translation="DYKDDDDK"
promoter complement(2144..2162)
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind complement(2145..2162)
/label=SP6
/note="SP6 promoter, forward primer"
primer_bind complement(2182..2199)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 2188..2412
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 2458..2886
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2545..2564)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 2755..2776
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(2895..2915)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 2900..3229
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 3080..3215
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 3142..3161
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 3296..4090
/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(3350..3369)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 3960..3979
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
polyA_signal 4264..4385
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(4301..4320)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 4355..4374
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(4434..4450)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4434..4450)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(4447..4469)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 4458..4474
/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(4482..4512)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4527..4548
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(4665..4682)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4836..5424)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4916..4935)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5595..6455)
/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 6218..6237
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(6456..6560)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga
61 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg
121 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct
181 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca
241 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata
301 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
361 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
421 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
481 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
541 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
601 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
661 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
721 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
781 cggtaggcgt gtacggtggg aggtctatat aagcagagct ctctggctaa ctagagaacc
841 cactgcttac tggcttatcg aaattaatac gactcactat agggagaccc aagcttcggc
901 agccaacatg gactataagg acgatgatga caaatctcag tcgaaaggca agaagcgaaa
961 ccctggcctt aaaattccaa aagaagcatt tgaacaacct cagaccagtt ccacaccacc
1021 tagagattta gactccaagg cttgcatttc tattggaaat cagaactttg aggtgaaggc
1081 agatgacctg gagcctataa tggaactggg acgaggtgcg tacggggtgg tggagaagat
1141 gcggcacgtg cccagcgggc agatcatggc agtgaagcgg atccgagcca cagtaaatag
1201 ccaggaacag aaacggctac tgatggattt ggatatttcc atgaggacgg tggactgtcc
1261 attcactgtc accttttatg gcgcactgtt tcgggagggt gatgtgtgga tctgcatgga
1321 gctcatggat acatcactag ataaattcta caaacaagtt attgataaag gccagacaat
1381 tccagaggac atcttaggga aaatagcagt ttctattgta aaagcattag aacatttaca
1441 tagtaagctg tctgtcattc acagagacgt caagccttct aatgtactca tcaatgctct
1501 cggtcaagtg aagatgtgcg attttggaat cagtggctac ttggtggact ctgttgctaa
1561 aacaattgat gcaggttgca aaccatacat ggcccctgaa agaataaacc cagagctcaa
1621 ccagaaggga tacagtgtga agtctgacat ttggagtctg ggcatcacga tgattgagtt
1681 ggccatcctt cgatttccct atgattcatg gggaactcca tttcagcagc tcaaacaggt
1741 ggtagaggag ccatcgccac aactcccagc agacaagttc tctgcagagt ttgttgactt
1801 tacctcacag tgcttaaaga agaattccaa agaacggcct acatacccag agctaatgca
1861 acatccattt ttcaccctac atgaatccaa aggaacagat gtggcatctt ttgtaaaact
1921 gattcttgga gactaaaaag cagtggactt aatcggttga ccctactgtg gattggtggg
1981 tttcggggtg aagcaagttc actacagcat caatagaaag tcatctttga gataatttaa
2041 ccctgcctct cagagggttt tctctcccaa ttttcttttt actccccctc ttaagggggc
2101 cttggaatct atagtataga atgaactgtc tagagggccc tattctatag tgtcacctaa
2161 atgctagagc tcgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt
2221 gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat
2281 aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg
2341 tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg
2401 tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg
2461 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
2521 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
2581 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
2641 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
2701 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
2761 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
2821 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
2881 cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
2941 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
3001 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
3061 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
3121 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct
3181 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg
3241 agcttgtata tccattttcg gatctgatca agagacagga tgaggatcgt ttcgcatgat
3301 tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta
3361 tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca
3421 ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga
3481 cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga
3541 cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct
3601 cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg
3661 gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga
3721 gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca
3781 tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga
3841 ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg
3901 cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc
3961 gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt
4021 gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga
4081 gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca
4141 tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc
4201 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac
4261 cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
4321 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
4381 tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag
4441 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
4501 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
4561 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
4621 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
4681 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
4741 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
4801 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
4861 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
4921 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
4981 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
5041 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
5101 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
5161 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
5221 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
5281 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
5341 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
5401 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
5461 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
5521 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
5581 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
5641 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
5701 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
5761 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
5821 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
5881 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
5941 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
6001 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
6061 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
6121 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
6181 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
6241 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
6301 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
6361 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
6421 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
6481 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
6541 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacg
//
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