- 规格:20μl质粒
- 启动子:CaMV 35S promoter
- 终止子:NOS terminator
- 质粒大小:10829bp
- 质粒标签:N-FLAG
- 原核抗性:卡那霉素Kan
- 筛选标记:潮霉素Hyg
- 克隆菌株:大肠杆菌DB3.1
- 质粒简介: pCXSN-FLAG质粒是一个植物过表达质粒,35S启动子驱动FLAG标签和目的基因融合表达,在ccdb致死基因的作用下,克隆基因变得更加容易,阳性率高。
- 质粒图谱1: png/1-1G0301P51V01.png
质粒序列: 点击查看序列LOCUS Exported 10829 bp ds-DNA circular SYN 30-OCT-2017DEFINITION synthetic circular DNASOURCE synthetic DNA constructORGANISM synthetic DNA constructREFERENCE 1 (bases 1 to 10829)AUTHORS .TITLE Direct Submissionhttp://www.fenghbio.cnFEATURES Location/Qualifierssource 1..10829/organism="synthetic DNA construct"/mol_type="other DNA"terminator 7..270/label=NOS terminator/note="nopaline synthase terminator and poly(A) signal"CDS 627..932/codon_start=1/gene="ccdB"/product="CcdB, a bacterial toxin that poisons DNA gyrase"/label=ccdB/note="Plasmids containing the ccdB gene cannot bepropagated in standard E. coli strains."/translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDKVSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"CDS complement(988..1011)/codon_start=1/product="FLAG(R) epitope tag, followed by an enterokinasecleavage site"/label=FLAG/translation="DYKDDDDK"promoter complement(1029..1919)/label=CaMV 35S promoter/note="strong constitutive promoter from cauliflower mosaicvirus"misc_feature 2153..2177/label=RB T-DNA repeat/note="right border repeat from nopaline C58 T-DNA"CDS 3476..4105/codon_start=1/product="stability protein from the Pseudomonas plasmidpVS1 (Heeb et al., 2000)"/label=pVS1 StaA/translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSARDWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIPVQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPILESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"CDS 4539..5606/codon_start=1/product="replication protein from the Pseudomonas plasmidpVS1 (Heeb et al., 2000)"/label=pVS1 RepA/translation="GRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESWQAAADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLSKRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKPGRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGEALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYRLARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPILVMRYRNLIEGEASAGS"rep_origin 5672..5866/label=pVS1 oriV/note="origin of replication for the Pseudomonas plasmidpVS1 (Heeb et al., 2000)"misc_feature 6210..6350/label=bom/note="basis of mobility region from pBR322"rep_origin complement(6536..7124)/direction=LEFT/label=ori/note="high-copy-number ColE1/pMB1/pBR322/pUC origin ofreplication"CDS complement(7211..8005)/codon_start=1/gene="aphA-3"/product="aminoglycoside phosphotransferase"/label=KanR/note="confers resistance to kanamycin"/translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKMTDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYEDEQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDTPFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIAFCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"misc_feature 8430..8454/label=LB T-DNA repeat/note="left border repeat from nopaline C58 T-DNA"polyA_signal 8532..8706/label=CaMV poly(A) signal/note="cauliflower mosaic virus polyadenylation signal"CDS complement(8746..9771)/codon_start=1/gene="aph(4)-Ia"/product="aminoglycoside phosphotransferase from E. coli"/label=HygR/note="confers resistance to hygromycin"/translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKK"promoter complement(9838..10515)/label=CaMV 35S promoter (enhanced)/note="cauliflower mosaic virus 35S promoter with aduplicated enhancer region"ORIGIN1 gaattccgat ctagtaacat agatgacacc gcgcgcgata atttatccta gtttgcgcgc61 tatattttgt tttctatcgc gtattaaatg tataattgcg ggactctaat cataaaaacc121 catctcataa ataacgtcat gcattacatg ttaattatta catgcttaac gtaattcaac181 agaaattata tgataatcat cgcaagaccg gcaacaggat tcaatcttaa gaaactttat241 tgccaaatgt ttgaacgatc ggggaaattc gctagtggat ccccaatact tgtatggggc301 ttactaaaag ccagataaca gtatgcgtat ttgcgcgctg atttttgcgg tataagaata361 tatactgata tgtatacccg aagtatgtca aaaagaggta tgctatgaag cagcgtatta421 cagtgacagt tgacagcgac agctatcagt tgctcaaggc atatatgatg tcaatatctc481 cggtctggta agcacaacca tgcagaatga agcccgtcgt ctgcgtgccg aacgctggaa541 agcggaaaat caggaaggga tggctgaggt cgcccggttt attgaaatga acggctcttt601 tgctgacgag aacaggggct ggtgaaatgc agtttaaggt ttacacctat aaaagagaga661 gccgttatcg tctgtttgtg gatgtacaga gtgatattat tgacacgccc gggcgacgga721 tggtgatccc cctggccagt gcacgtctgc tgtcagataa agtctcccgt gaactttacc781 cggtggtgca tatcggggat gaaagctggc gcatgatgac caccgatatg gccagtgtgc841 cggtctccgt tatcggggaa gaagtggctg atctcagcca ccgcgaaaat gacatcaaaa901 acgccattaa cctgatgttc tggggaatat aaatgtcagg ctcccttata cacagccagt961 ctgcaggtcg acccatacaa gtattggctt atcatcatca tccttgtaat ccatggatcc1021 ccctcgagta tcgttcgtaa atggtgaaaa ttttcagaaa attgcttttg ctttaaaaga1081 aatgatttaa attgctgcaa tagaagtaga atgcttgatt gcttgagatt cgtttgtttt1141 gtatatgttg tgttgaggtc gaggtcctct ccaaatgaaa tgaacttcct tatatagagg1201 aagggtcttg cgaaggatag tgggattgtg cgtcatccct tacgtcagtg gagatatcac1261 atcaatccac ttgctttgaa gacgtggttg gaacgtcttc tttttccacg atgctcctcg1321 tgggtggggg tccatctttg ggaccactgt cggcagaggc atcttcaacg atggcctttc1381 ctttatcgca atgatggcat ttgtaggagc caccttcctt ttccactatc ttcacaataa1441 agtgacagat agctgggcaa tggaatccga ggaggtttcc ggatattacc ctttgttgaa1501 aagtctcaat tgccctttgg tcttctgaga ctgtatcttt gatatttttg gagtagacaa1561 gtgtgtcgtg ctccaccatg ttatcacatc aatccacttg ctttgaagac gtggttggaa1621 cgtcttcttt ttccacgatg ctcctcgtgg gtgggggtcc atctttggga ccactgtcgg1681 cagaggcatc ttcaacgatg gcctttcctt tatcgcaatg atggcatttg taggagccac1741 cttccttttc cactatcttc acaataaagt gacagatagc tgggcaatgg aatccgagga1801 ggtttccgga tattaccctt tgttgaaaag tctcaattgc cctttggtct tctgagactg1861 tatctttgat atttttggag tagacaagtg tgtcgtgctc caccatgttg acctgcaggc1921 atgcaagctt ggcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta1981 cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg2041 cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgc tagagcagct2101 tgagcttgga tcagattgtc gtttcccgcc ttcagtttaa actatcagtg tttgacagga2161 tatattggcg ggtaaaccta agagaaaaga gcgtttatta gaataacgga tatttaaaag2221 ggcgtgaaaa ggtttatccg ttcgtccatt tgtatgtgca tgccaaccac agggttcccc2281 tcgggatcaa agtactttga tccaacccct ccgctgctat agtgcagtcg gcttctgacg2341 ttcagtgcag ccgtcttctg aaaacgacat gtcgcacaag tcctaagtta cgcgacaggc2401 tgccgccctg cccttttcct ggcgttttct tgtcgcgtgt tttagtcgca taaagtagaa2461 tacttgcgac tagaaccgga gacattacgc catgaacaag agcgccgccg ctggcctgct2521 gggctatgcc cgcgtcagca ccgacgacca ggacttgacc aaccaacggg ccgaactgca2581 cgcggccggc tgcaccaagc tgttttccga gaagatcacc ggcaccaggc gcgaccgccc2641 ggagctggcc aggatgcttg accacctagc cctggcgacg ttgtgacagt gaccaggcta2701 gaccgcctgg cccgcagcac ccgcgaccta ctggacattg ccgagcgcat ccaggaggcc2761 ggcgcgggcc tgcgtagcct ggcagagccg tgggccgaca ccaccacgcc ggccggccgc![]()