| 质粒名称: | RN3P-KDM5B-FLAG |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 86398 |
| 存储实验室: | Wei Xie |
|
载体骨架基本信息 |
|
| 载体名称: | RN3P |
| 载体出品公司: | Derivative of pBluescript RN3 |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | -- |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 哺乳细胞表达 |
| 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: | KDM5B |
| 物种来源: | M. musculus (mouse) |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | Lac |
| 标签/融合蛋白: | 3x-FLAG（主干上的N终端） |
| 克隆方法: | Gibson克隆 |
| 5' 克隆位点: | -- |
| 3' 克隆位点: | -- |
| 5' 测序引物 | M13 Reverse |
| 3' 测序引物 | XBG-R (GACTCCATTCGGGTGTTC) |
|
文献引用方法 |
|
| 材料和方法部分: | RN3P-KDM5B-FLAG was a gift from Wei Xie (Addgene plasmid # 86398 ; http://n2t.net/addgene:86398 ; RRID:Addgene_86398) |
| 参考文献部分: | Allelic reprogramming of the histone modification H3K4me3 in early mammalian development. Zhang B, Zheng H, Huang B, Li W, Xiang Y, Peng X, Ming J, Wu X, Zhang Y, Xu Q, Liu W, Kou X, Zhao Y, He W, Li C, Chen B, Li Y, Wang Q, Ma J, Yin Q, Kee K, Meng A, Gao S, Xu F, Na J, Xie W. Nature. 2016 Sep 14;537(7621):553-557. doi: 10.1038/nature19361. 10.1038/nature19361 PubMed 27626382 |
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 RN3P-KDM5B-FLAG 7915 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION FLAG-tagged mouse KDM5B cDNA clone for mammalian cell expression.
ACCESSION .
VERSION .
KEYWORDS RN3P-KDM5B-FLAG
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7915)
AUTHORS Zhang B, Zheng H, Huang B, Li W, Xiang Y, Peng X, Ming J, Wu X,
Zhang Y, Xu Q, Liu W, Kou X, Zhao Y, He W, Li C, Chen B, Li Y, Wang
Q, Ma J, Yin Q, Kee K, Meng A, Gao S, Xu F, Na J, Xie W
TITLE Allelic reprogramming of the histone modification H3K4me3 in early
mammalian development.
JOURNAL Nature. 2016 Sep 14;537(7621):553-557. doi: 10.1038/nature19361.
PUBMED 27626382
REFERENCE 2 (bases 1 to 7915)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..7915
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 432..451
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 685..702
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 819..840
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 855..885
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 893..909
/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)."
primer_bind 898..920
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 917..933
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 917..933
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
promoter 954..972
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
5'UTR 997..1039
/label=Xenopus globin 5'-UTR
/note="translational enhancer from the 5'-UTR of the major
beta-globin gene of Xenopus laevis"
CDS 1069..1134
/codon_start=1
/product="three tandem FLAG(R) epitope tags, followed by an
enterokinase cleavage site"
/label=3xFLAG
/translation="DYKDHDGDYKDHDIDYKDDDDK"
3'UTR 5841..5966
/label=Xenopus globin 3'-UTR
/note="3'-UTR of the major beta-globin gene of Xenopus
laevis"
primer_bind complement(6056..6075)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(6057..6075)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(6082..6099)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(6082..6098)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(6091..6113)
/label=M13/pUC Forward
/note="In lacZ gene"
rep_origin complement(6240..6695)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(6377..6398)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 6589..6608
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 6722..6826
/gene="bla"
/label=AmpR promoter
CDS 6827..7687
/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 complement(7045..7064)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 7858..531
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
ORIGIN
1 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
61 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
121 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
181 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
241 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
301 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
361 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
421 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
481 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
541 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
601 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
661 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca
721 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg
781 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat
841 taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc
901 ggataacaat ttcacacagg aaacagctat gaccatgatt acgccaagct cgaaattaac
961 cctcactaaa gggaacaaaa gctggagcta agcttgcttg ttctttttgc agaagctcag
1021 aataaacgct caactttggc agatctgaat tcctgcagcc cggggatgga ctacaaagac
1081 catgacggtg attataaaga tcatgatatc gattacaagg atgacgatga caagggatct
1141 gcaagaagag catctgtgga attcctgcag cccgggtcca tggagccggc caccacgctg
1201 cccccaggcc cgcgcccggc gctgcccctc gggggcccgg gcccgctggg agagttcctg
1261 cctccccccg agtgtccggt cttcgagccg agctgggaag agttcgcgga ccccttcgct
1321 ttcatccaca agatccggcc catagccgag cagactggga tctgtaaggt gcggccgccg
1381 ccggattggc agccaccatt cgcttgtgat gtcgataaac ttcatttcac cccgcggatc
1441 cagaggctga atgagctgga ggcccaaaca cgtgtaaaat tgaatttcct ggaccagatt
1501 gcaaagtatt gggagctaca aggaagtact ctgaagattc cccacgtgga gagaaagatc
1561 ttagacttgt tccagctaaa taagttagtc gcagaagaag gtgggtttgc agttgtttgc
1621 aaggatagaa aatggaccaa aattgccacc aagatgggtt ttgctcctgg taaagctgtg
1681 ggctcacata tcaggggaca ttatgagcga atactcaatc catacaattt gttcctgtct
1741 ggagacagtc tgaggtgtct tcagaagcct aacctgacct ccgacacaaa ggacaaggag
1801 tacaaacccc atgatattcc ccagaggcag tctgtgcagc ccgcagaaac ctgtcctccg
1861 gcccgccgag caaagcgcat gagagcagag gctatgaata ttaaaattga gccagaagag
1921 gcaacagaag ccaggactca taatctgaga cgccgaatgg gttgcacaac tccaaaatgg
1981 gaaaatgaga aagaaatgaa aagcaccatc aagcaagagc ccactgagaa gaaagactgc
2041 gagttagaaa gtgagaagga gaagcccaag agccgggcca agaaaacggc cactgcggtg
2101 gacctgtatg tgtgcctctt atgtggcagt ggcaatgacg aggaccggct cctactgtgt
2161 gacggctgtg atgacagtta ccacaccttc tgcctggttc cccctctcca tgacgttccc
2221 aagggagact ggaggtgtcc taagtgtttg gctcaagaat gtaataagcc acaagaagct
2281 tttggctttg aacaagcagc aagggactat acacttcgta cgtttggaga aatggcagat
2341 gcattcaaat ctgattactt caacatgcca gtccatatgg ttcccacaga gctggtggag
2401 aaggagtttt ggcggctcgt aagcactatt gaagaagacg tcacggtgga gtatggagct
2461 gatattgctt caaaagagtt tggcagtggc tttcctgttc gagatgggaa aatcaagatt
2521 tctcctgagg aagaggaata cctggatagt ggctggaatt tgaacaacat gccagtgatg
2581 gagcagtctg tccttgctca tattactgct gatatatgtg gcatgaaact cccctggttg
2641 tatgtgggca tgtgcttttc ttcattctgt tggcacattg aagaccactg gagctattca
2701 atcaactacc tacactgggg tgagccaaaa acctggtatg gagttccagg ctacgctgct
2761 gagcagctag aaaacgtgat gaagaagctc gctcctgagc tctttgtgtc ccagccggac
2821 ctgctccacc agctggtgac catcatgaac cccaacactc tgatgactca tgaagtgcct
2881 gtttatcgaa cgaatcagtg tgctggggag tttgtaatta cgtttccaag agcctaccac
2941 agtggcttca accaaggttt taattttgcc gaggctgtta acttctgcac tgttgattgg
3001 ctgccattgg gccgccagtg tgtggagcat tatcgcttgc tgcaccgtta ttgtgtcttt
3061 tcccacgatg aaatgatatg caaaatggct tctaaggctg atgtattaga cgttgtggta
3121 gcgtcaactg ttcagaagga catggccatc atgatcgagg acgagaaagc tttaagggaa
3181 actgttcgga aattgggagt aattgattct gaaagaatgg attttgagct gttgccagat
3241 gatgagcggc agtgtataaa atgcaaaact acatgcttca tgtctgccat ctcctgctct
3301 tgtaaacctg gcctacttgt ttgcctacat catgtgaaag aattgtgttc ttgccccccg
3361 tataagtata acctgcggta tagatacact ctggatgatc tctatccaat gatgaatgca
3421 ttaaagcttc gagcagagtc ttacaatgaa tgggccttaa atgtgaatga agccttggaa
3481 gcaaagatca ataagaagaa aagtcttgtc agctttaaag ctttgattga agaatcagaa
3541 atgaagaaat tcccagacaa tgatcttttg cgtcaccttc gactagtcac gcaggatgca
3601 gaaaagtgtg cctctgttgc ccagcagttg cttaatggca aaaggcaaac aaggtatcga
3661 tctggtggag gaaagtctca gaatcagctg acggtgaatg aacttagaca gtttgtgact
3721 cagctgtatg ctctgccgtg cgtccttagt cagacaccgt tgctaaagga tctcttgaat
3781 cgtgtggaag atttccaaca gcaaagccaa aaactactgt ctgaggaaat gcctagcgct
3841 gccgagctac aggagttgct ggatgtcagc tttgaatttg atgttgagct tccacagctc
3901 actgagatgc gcattcgctt agagcaggcc cgctggctgg aagaggttca gcaggcttgc
3961 ctggactcca gctccctttc tttggatgat atgagacgac tcatagactt aggggtgggg
4021 ctggctccat attctgctgt ggagaaagct atggcccgcc tgcaggagct gctaacagtg
4081 tcagaacact gggacgacaa agccaagagt ctcctcagag caagaccacg gcattctttg
4141 agtagccttg ctacagcagt caaagagatg gaggagatcc ccgcctacct gcccaatggt
4201 acagtcctga aggattctgt gcaaagagcc agagactggg ttcaggatgt ggatgcattg
4261 caggcaggag gacgtgtgcc agtgttagag acactcattg aacttgttgc aagaggccga
4321 tctatcccag tacatctcaa ttctttacca agattggaaa tgttagtagc tgaagttcat
4381 gcatggaaag aatgtgctgc taaaacattc ttgcctgaga attctacata ttctctgtta
4441 gaggttcttt gccctcggtg tgacattggc cttttggggc tgaaaaggaa acagagaaag
4501 ttgaaggagc ccttgccgag tggaaagaag agaagcacca agttagagag cctgagcgat
4561 ctggagaggg ctctgatgga gagcaaagag actgcggcag ctatggcaac tcttggggaa
4621 gctcgcctac gagagatgga agctttgcag tctctcagat ttgccaatga agagaaattg
4681 ctgtcacctg tccaagactt agagatgaaa gtctgcttgt gtcagaagac tccagccact
4741 cccatgattc agtgtgaact ctgcagggat gctttccata ctagttgtgt ggcagcaccc
4801 agtatttcac aaagctcccg aatctggctt tgtcctcact gtcggaggtc agagaagcct
4861 ccattagaga aaattctgcc tctgctggcc tccctacagc gtatacgagt tcgtcttcct
4921 gaaggagatg cactccgata catgattgaa agaaccgtga actggcaaca tagagcccag
4981 cagctgcttt cttcagggaa tctaaaactt gtgcaagacc aagtgggttc tggactgttg
5041 tctagcagat ggccagcctc agcaggacag gcatcagcaa cagacaaggt gtctcagcct
5101 cctggcacaa catccttttc attgcccgat gactgggaca acagaacctc atatttacac
5161 tctcccttct ctactggaca gagttgtctc cccctccatg gtttgagtcc agaagtgaat
5221 gaattgctga tggaagccca gctgctccag gtatcccttc ctgaaattca ggagctttat
5281 cagactctac ttactaagcc aagctctgtt cagcaagctg accgaagttc accagttaga
5341 tccagcagtg agaagaacga ttgccttcga gggaaacgag atgcaatcaa cagtcctgag
5401 aggaaactga aaaggcgccc agaaagagaa ggccttccta gcgagcggtg ggatcgagtt
5461 aaacacatgc ggacccccca aaagaagaaa atcaaactga gtcaccccaa ggacatggac
5521 agtttcaagt tagaaagaga gcgtagctat gacttagtgc gtaatgctga aactcattcc
5581 ctgccctcag acacatccta ctccgaacag gaggactctg aggatgaaga tgccatctgt
5641 ccagctgtga gctgcctgca gccagaagga gatgaggtgg actgggtcca gtgtgatggc
5701 agctgcaatc agtggtttca ccaggtctgt gttggtgtct ctccagagat ggcagagaag
5761 gaagactaca tctgtgtgcg ctgtactggg aaggacgcac caagccgaaa gtaaagcggc
5821 cgcggatctg gttaccacta aaccagcctc aagaacaccc gaatggagtc tctaagctac
5881 ataataccaa cttacacttt acaaaatgtt gtcccccaaa atgtagccat tcgtatctgc
5941 tcctaataaa aagaaagttt cttcacattc taaaaaaaaa aaaaaaaaaa aaaacccccc
6001 cccccccccc cccccccccc cccctgcagg gccaagtcgg ccggtaccca attcgcccta
6061 tagtgagtcg tattacaatt cactggccgt cgttttacaa cgtcgtgact gggaaaaccc
6121 tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag
6181 cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggca
6241 aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt
6301 ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat
6361 agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa
6421 cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta
6481 atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc
6541 ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc
6601 gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac
6661 acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt
6721 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
6781 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
6841 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
6901 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
6961 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
7021 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
7081 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
7141 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
7201 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
7261 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
7321 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
7381 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
7441 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
7501 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
7561 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
7621 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
7681 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
7741 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
7801 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
7861 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgcta
//
