| 质粒名称: | pClneoHA mouse Numb |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 37012 |
| 存储实验室: | Yutaka Hata |
|
载体骨架基本信息 |
|
| 载体名称: | pCl-neo |
| 载体出品公司: | Promega |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 5472 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: | Numb |
| 物种来源: | M. musculus (mouse) |
| 插入大小: | -- |
| 基因突变: | contains Numb aa 8-853 |
| GenBank编号 | BC033459 |
| 启动子 | CMV |
| 标签/融合蛋白: | HA（插入件上的N端子） |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | EcoRI(未破坏) |
| 3' 克隆位点: | SalI(未破坏) |
| 5' 测序引物 | CMV-F |
| 3' 测序引物 | T3 |
|
文献引用方法 |
|
| 材料和方法部分: | pClneoHA mouse Numb was a gift from Yutaka Hata (Addgene plasmid # 37012 ; http://n2t.net/addgene:37012 ; RRID:Addgene_37012) |
| 参考文献部分: | Ligand-of-Numb protein X is an endocytic scaffold for junctional adhesion molecule 4. Kansaku A, Hirabayashi S, Mori H, Fujiwara N, Kawata A, Ikeda M, Rokukawa C, Kurihara H, Hata Y. Oncogene. 2006 Aug 24;25(37):5071-84. Epub 2006 Jul 10. 10.1038/sj.onc.1209468 PubMed 16832352 |
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 IgK-IFN-luc 5863 bp ds-DNA circular SYN 19-MAR-2019
DEFINITION Reporter construct for NF‐kappa-B activation. Contains two copies of
immunoglobulin kappa light chain motif (5′‐GGGGACTTTCC‐3′) upstream
of the a minimal promoter driving luciferase expression.
ACCESSION .
VERSION .
KEYWORDS IgK-IFN-luc
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5863)
AUTHORS Pomerantz JL, Denny EM, Baltimore D
TITLE CARD11 mediates factor-specific activation of NF-kappaB by the T
cell receptor complex.
JOURNAL EMBO J. 2002 Oct 1. 21(19):5184-94.
PUBMED 12356734
REFERENCE 2 (bases 1 to 5863)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Mar 19, 2019 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..5863
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter complement(33..51)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(72..88)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(72..88)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(85..107)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 96..112
/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(120..150)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 165..186
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(303..320)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(474..1062)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(554..573)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(1233..2093)
/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 1856..1875
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(2094..2198)
/gene="bla"
/label=AmpR promoter
rep_origin complement(2224..2679)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2361..2382)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 2573..2592
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 2806..2828
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 2820..2837
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 2821..2837
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 2844..2863
/label=T7
/note="T7 promoter, forward primer"
promoter 2844..2862
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
polyA_signal 2872..3006
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
CDS 3210..4862
/codon_start=1
/gene="luc"
/product="firefly luciferase"
/label=luciferase
/translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
HIEVNITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
ANDIYNERELLNSMNISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
FVDEVPKGLTGKLDARKIREILIKAKKGGKSKL"
primer_bind complement(3276..3294)
/label=LucNrev
/note="Firefly luciferase, reverse primer"
intron 5086..5151
/label=small t intron
/note="SV40 (simian virus 40) small t antigen intron"
CDS 5281..5301
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
polyA_signal 5726..5860
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
ORIGIN
1 ctcgaggggg ggcccggtac cagcttttgt tccctttagt gagggttaat ttcgagcttg
61 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
121 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
181 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
241 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
301 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
361 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
421 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
481 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
541 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
601 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
661 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
721 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
781 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
841 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
901 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
961 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
1021 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
1081 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
1141 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
1201 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
1261 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
1321 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
1381 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
1441 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
1501 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
1561 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
1621 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
1681 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
1741 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
1801 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
1861 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
1921 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
1981 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
2041 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
2101 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
2161 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
2221 cctaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct
2281 cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg
2341 agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact
2401 ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac
2461 cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga
2521 gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga
2581 aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca
2641 ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc ccattcgcca ttcaggctgc
2701 gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag
2761 ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt
2821 gtaaaacgac ggccagtgaa ttgtaatacg actcactata gggcgaattg gaacttgttt
2881 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
2941 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
3001 tggatcgatc cccccaccgc ggtggcggcc cacagagggg actttccgag agatctacag
3061 aggggacttt ccgagagcga gcttgggctg caggtcgaga tccgctgaat agagagagga
3121 ccatctcata taaataggcc atacccatgg agaaaggaca ttctaactgc aacctttcgc
3181 aagcttggca ttccggtact gttggtaaaa tggaagacgc caaaaacata aagaaaggcc
3241 cggcgccatt ctatcctcta gaggatggaa ccgctggaga gcaactgcat aaggctatga
3301 agagatacgc cctggttcct ggaacaattg cttttacaga tgcacatatc gaggtgaaca
3361 tcacgtacgc ggaatacttc gaaatgtccg ttcggttggc agaagctatg aaacgatatg
3421 ggctgaatac aaatcacaga atcgtcgtat gcagtgaaaa ctctcttcaa ttctttatgc
3481 cggtgttggg cgcgttattt atcggagttg cagttgcgcc cgcgaacgac atttataatg
3541 aacgtgaatt gctcaacagt atgaacattt cgcagcctac cgtagtgttt gtttccaaaa
3601 aggggttgca aaaaattttg aacgtgcaaa aaaaattacc aataatccag aaaattatta
3661 tcatggattc taaaacggat taccagggat ttcagtcgat gtacacgttc gtcacatctc
3721 atctacctcc cggttttaat gaatacgatt ttgtaccaga gtcctttgat cgtgacaaaa
3781 caattgcact gataatgaat tcctctggat ctactgggtt acctaagggt gtggcccttc
3841 cgcatagaac tgcctgcgtc agattctcgc atgccagaga tcctattttt ggcaatcaaa
3901 tcattccgga tactgcgatt ttaagtgttg ttccattcca tcacggtttt ggaatgttta
3961 ctacactcgg atatttgata tgtggatttc gagtcgtctt aatgtataga tttgaagaag
4021 agctgttttt acgatccctt caggattaca aaattcaaag tgcgttgcta gtaccaaccc
4081 tattttcatt cttcgccaaa agcactctga ttgacaaata cgatttatct aatttacacg
4141 aaattgcttc tgggggcgca cctctttcga aagaagtcgg ggaagcggtt gcaaaacgct
4201 tccatcttcc agggatacga caaggatatg ggctcactga gactacatca gctattctga
4261 ttacacccga gggggatgat aaaccgggcg cggtcggtaa agttgttcca ttttttgaag
4321 cgaaggttgt ggatctggat accgggaaaa cgctgggcgt taatcagaga ggcgaattat
4381 gtgtcagagg acctatgatt atgtccggtt atgtaaacaa tccggaagcg accaacgcct
4441 tgattgacaa ggatggatgg ctacattctg gagacatagc ttactgggac gaagacgaac
4501 acttcttcat agttgaccgc ttgaagtctt taattaaata caaaggatat caggtggccc
4561 ccgctgaatt ggaatcgata ttgttacaac accccaacat cttcgacgcg ggcgtggcag
4621 gtcttcccga cgatgacgcc ggtgaacttc ccgccgccgt tgttgttttg gagcacggaa
4681 agacgatgac ggaaaaagag atcgtggatt acgtcgccag tcaagtaaca accgcgaaaa
4741 agttgcgcgg aggagttgtg tttgtggacg aagtaccgaa aggtcttacc ggaaaactcg
4801 acgcaagaaa aatcagagag atcctcataa aggccaagaa gggcggaaag tccaaattgt
4861 aaaatgtaac tgtattcagc gatgacgaaa ttcttagcta ttgtaatatt atatgcaaat
4921 tgatgaatgg taattttgta attgtgggtc actgtactat tttaacgaat aataaaatca
4981 ggtataggta actaaaaagg aattcggggg atctttgtga aggaacctta cttctgtggt
5041 gtgacataat tggacaaact acctacagag atttaaagct ctaaggtaaa tataaaattt
5101 ttaagtgtat aatgtgttaa actactgatt ctaattgttt gtgtatttta gattccaacc
5161 tatggaactg atgaatggga gcagtggtgg aatgccttta atgaggaaaa cctgttttgc
5221 tcagaagaaa tgccatctag tgatgatgag gctactgctg actctcaaca ttctactcct
5281 ccaaaaaaga agagaaaggt agaagacccc aaggactttc cttcagaatt gctaagtttt
5341 ttgagtcatg ctgtgtttag taatagaact cttgcttgct ttgctattta caccacaaag
5401 gaaaaagctg cactgctata caagaaaatt atggaaaaat attctgtaac ctttataagt
5461 aggcataaca gttataatca taacatactg ttttttctta ctccacacag gcatagagtg
5521 tctgctatta ataactatgc tcaaaaattg tgtaccttta gctttttaat ttgtaaaggg
5581 gttaataagg aatatttgat gtatagtgcc ttgactagag atcataatca gccataccac
5641 atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca
5701 taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata
5761 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
5821 tttgtccaaa ctcatcaatg tatcttatca tgtctggatc gac
//
