| 质粒名称: | 3XFlagNICD1 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 20183 |
| 存储实验室: | Raphael Kopan |
|
载体骨架基本信息 |
|
| 载体名称: | p3XFLAG-CMV-7 |
| 载体出品公司: | Sigma |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 4700 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: | -- |
| 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: | Notch 1 Intracellular Domain,NICD1,Notch 1 |
| 物种来源: | M. musculus (mouse) |
| 插入大小: | 2361 bp |
| 基因突变: | 包含小鼠Notch 1细胞内结构域（Val-1744至Lys-2531） |
| GenBank编号 | AAM28905.1 AF508809_1 |
| 启动子 | CMV |
| 标签/融合蛋白: | 3XFLAG（主干网上的N个终端） |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | Xba1(未破坏) |
| 3' 克隆位点: | BamH1 (未破坏) |
| 5' 测序引物 | CMV-F |
| 3' 测序引物 | hGH-pA-R |
|
文献引用方法 |
|
| 材料和方法部分: | 3XFlagNICD1 was a gift from Raphael Kopan (Addgene plasmid # 20183 ; http://n2t.net/addgene:20183 ; RRID:Addgene_20183) |
| 参考文献部分: | Target selectivity of vertebrate notch proteins. Collaboration between discrete domains and CSL-binding site architecture determines activation probability. Ong CT, Cheng HT, Chang LW, Ohtsuka T, Kageyama R, Stormo GD, Kopan R. J Biol Chem. 2006 Feb 24. 281(8):5106-19. 10.1074/jbc.M506108200 PubMed 16365048 |
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 3XFlagNICD1 7057 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS 3XFlagNICD1
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7057)
AUTHORS Ong CT, Cheng HT, Chang LW, Ohtsuka T, Kageyama R, Stormo GD, Kopan
R
TITLE Target selectivity of vertebrate notch proteins. Collaboration
between discrete domains and CSL-binding site architecture
determines activation probability.
JOURNAL J Biol Chem. 2006 Feb 24. 281(8):5106-19.
PUBMED 16365048
REFERENCE 2 (bases 1 to 7057)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..7057
/organism="synthetic DNA construct"
/mol_type="other DNA"
protein_bind 107..128
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 143..173
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 181..197
/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 186..208
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 205..221
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 205..221
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind 235..254
/label=T7
/note="T7 promoter, forward primer"
promoter 235..253
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
promoter complement(292..621)
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 306..441
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind complement(360..379)
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
primer_bind 606..626
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
polyA_signal 650..1272
/label=hGH poly(A) signal
/note="human growth hormone polyadenylation signal"
primer_bind 1054..1073
/label=hGH-PA-R
/note="Human growth hormone terminator, reverse primer"
CDS complement(3674..3739)
/codon_start=1
/product="three tandem FLAG(R) epitope tags, followed by an
enterokinase cleavage site"
/label=3xFLAG
/translation="DYKDHDGDYKDHDIDYKDDDDK"
promoter complement(3769..3972)
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind complement(3798..3818)
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
enhancer 3973..4352
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
primer_bind complement(4513..4530)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(4513..4529)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4522..4544)
/label=M13/pUC Forward
/note="In lacZ gene"
rep_origin complement(4670..5125)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(4807..4828)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 5019..5038
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 5152..5256
/gene="bla"
/label=AmpR promoter
CDS 5257..6117
/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(5475..5494)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 6288..6876
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 6777..6796
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 7030..7047
/label=L4440
/note="L4440 vector, forward primer"
ORIGIN
1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
181 ttgtgagcgg ataacaattt cacacaggaa acagctatga catgattacg aatttaatac
241 gactcactat agggaattaa ctttctggtt tttcagttcc tcgaggagct ttttgcaaaa
301 gcctaggcct ccaaaaaagc ctcctcacta cttctggaat agctcagagg ccgaggcggc
361 ctcggcctct gcataaataa aaaaaattag tcagccatgg ggcggagaat gggcggaact
421 gggcggagtt aggggcggga tgggcggagt taggggcggg actatggttg ctgactaatt
481 gagatgcatg ctttgcatac ttctgcctgc tggggagcct ggggactttc cacacctggt
541 tgctgactaa ttgagatgca tgctttgcat acttctgcct gctggggagc ctggggactt
601 tccacaccct aactgacaca cattccacag ccaagctggc cgcgtaccca attcaacagg
661 catctactga gtggacccaa cgcatgagag gacagtgcca agcaagcaac tcaaatgtcc
721 caccggttgg gcatggccag gtagcctatg ctgtgtctgg acgtcctcct gctggtatag
781 ttattttaaa atcagaagga cagggaaggg agcagtggtt cacgcctgta atcccagcaa
841 tttgggaggc caaggtgggt agatcacctg agattaggag ttggagacca gcctggccaa
901 tatggtgaaa ccccgtctct accaaaaaaa caaaaattag ctgagcctgg tcatgcatgc
961 ctggaatccc aacaactcgg gaggctgagg caggagaatc gcttgaaccc aggaggcgga
1021 gattgcagtg agccaagatt gtgccactgc actccagctt ggttcccaat agaccccgca
1081 ggccctacag gttgtcttcc caacttgccc cttgctccat accacccccc tccaccccat
1141 aatattatag aaggacacct agtcagacaa aatgatgcaa cttaatttta ttaggacaag
1201 gctggtgggc actggagtgg caacttccag ggccaggaga ggcactgggg aggggtcaca
1261 gggatgccac ccgggatcct tatttaaatg cctctggaat gtgggtgatc tgggacggca
1321 tggtggtggg cgggctggag atgccctcgg accaatcaga gatgttggaa tgcggggagg
1381 agctggacca ctggtcaggg gactcagggg atggggtgag gaaggggtgc tctggcacct
1441 gcagctggtg gctgggggtg ttgtccacag gggaggagga gtaactgtgc tgggaaggag
1501 gggtcaggaa ctgggtagtg gtcatgggtg ggaccatgga ggatggcagt gatgtgggca
1561 gggcctggct ttcctggggc agaatggtgt gcacaggcag actgctgggg cccagcggtt
1621 gtacatctgc ctgactgggc tccccactca agaagctccg gcccaggtgc ccattggctg
1681 ccgagctcac actgaggtgt ggctgtgatg gtggctgcag gttctgaggc tggagctgta
1741 agttctgtgg ctgcacctgc tgggtctgca ccaggtgagg ctgtgttgcc agccgtgtgt
1801 tgggcaggcc ctggtaaata atcggggaca aggtattggt ggagaggctg ctgtgtagtg
1861 gccccatcat gctatgctgg aggccagctg cctgtgtgct cagtgtgccc ggcgtcacac
1921 ccggccgtag tgggttgtac tggctgggca ccatgccatt ctggagccgg ggaagccact
1981 cacactggcc attcaagctt gccggtgcac ccacggtgaa attcatagcc cccgtgccat
2041 tggtactcag cactgtgctg gcactggagg ctacaggcag gtgggagagg cgtggcgggg
2101 gtggctcaaa ggccaaccgg ctacctccag ccagtgctgc catctcaggc ttggctgcca
2161 cattcaagtg gctgatgccc aggtgagtgt caggcatacc aggcaggtgg ctgagaggca
2221 tggatggaga ctgctggaat ggggagggga ggaggggtgg cgaggccaca tctgacaagt
2281 agccatgggg tgactcgagg gagtccacag gcgacagcat gctcgagctg tccaacaggc
2341 agcccttgcc atcctgggac ttcttcctcc gtgccttgag gtccttagct tccttgctac
2401 cacaagccag ccctttggtg ctgggcttgc gggccttctt gccctgtgtg gcagacttga
2461 gattgcccag gtagccattg ggcgagcaga gtgtgggaga cagagtgggt gtgccaccca
2521 gggcagtgcc atgcagctgt gggctgcgca ccaggttgta ctcatccaaa agccgcacga
2581 tatcgtggtg catacgctcc tgtgcgatgt cccgcggcaa tcggtccatg tgatccgtga
2641 tgtcccggtt ggcaaagtgg tccagcaaca ctttggcagt ctcatagctg ccctcacggg
2701 cggccaggaa caggggagtc tcctccttgt tgttctgcat gtccttgttg gctccgttct
2761 tcaggagcac aacagcagca tccacattgt tcaccgcggc cgcccaatgc aaagccgact
2821 tgcctaggtc atccacggca ttgacgtcag catgtgagtt gatgaggtcc tccagcatgc
2881 cctccagggc caggcgcgca gccaggatca gtggagttgt gccatcatgc attcgggcat
2941 ccagatctgt ggccctgttc cggagcagga tctggaagac accctgagca tctgcagaaa
3001 ctgctgcatg taacggagta cggcccatgt tgtcctggat gttggcatct gcactggcct
3061 ccagcaagcg ctttgcagca tctgaacgag agtatcgggc agccaagtgc aaggcggtct
3121 ccccggtgcg gtctgtctgg ttgtgcaagc tggcgccctg gtagatgaag tcagagatga
3181 cagcaggtgc atcttcttct tcttcactgt tgcctgtctc aaggccccct ccactgcagg
3241 aggcaatcat gaggggtgtg aagccatctg gtcctcgaac attgacatcc atgcagtcag
3301 catccacctc cccctgaggc ggtgttgggg ccatggcaga catgcgcagg tcagcagcgt
3361 ccaggtgctg ctgggtccac tgtctgtggt cagtctgatc actcaggtca gggagaacta
3421 ctggctcctc aaaccggaac ttcttggtct ccaggtcttc gtctccccac tcgttctgat
3481 tgtcgtccat cagagcacca tctgaggcat tcttcagggg cttgaggccg actgagtcct
3541 cgccgagggg ctctctccgc ttcttcttgc tggcctctga cactttgaaa ccctcaggga
3601 accagagctg gccatgctgc cgccggcgct tgcgggacag cagcactcta gagtcgacct
3661 gcaggcatgc aagcttgtca tcgtcatcct tgtaatcgat atcatgatct ttataatcac
3721 cgtcatggtc tttgtagtcc atggtgaatt ctgacggttc actaaacgag ctctgcttat
3781 atagacctcc caccgtacac gcctaccgcc catttgcgtc aacggggcgg ggttattacg
3841 acattttgga aagtcccgtt gattttggtg ccaaaacaaa ctcccattga cgtcaatggg
3901 gtggagactt ggaaatcccc gtgagtcaaa ccgctatcca cgcccattgg tgtactgcca
3961 aaaccgcatc accatggtaa tagcgatgac taatacgtag atgtactgcc aagtaggaaa
4021 gtcccgtaag gtcatgtact gggcataatg ccaggcgggc catttaccgt cattgacgtc
4081 aatagggggc ggacttggca tatgatacac ttgatgtact gccaagtggg cagtttaccg
4141 taaatactcc acccattgac gtcaatggaa agtccctatt ggcgttacta tgggaacata
4201 cgtcattatt gacgtcaatg ggcgggggtc gttgggcggt cagccaggcg ggccatttac
4261 cgtaagttat gtaacgcgga actccatata tgggctatga actaatgacc ccgtaattga
4321 ttactattaa taactagtca ataatcaatg tcaacatggc ggtcatattg gacatgagcc
4381 aatataaatg tacatattat gatatagata caacgtatgc aatggccaat agccaatatt
4441 gatttatgct atataaccaa tgactaatat ggctaattgc caatattgat tcaatgtata
4501 gatcagcttg gcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac
4561 ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc
4621 ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatgga aattgtaagc
4681 gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa
4741 taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt
4801 gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg
4861 cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt
4921 ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga
4981 gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg
5041 ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg
5101 cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
5161 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
5221 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
5281 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
5341 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
5401 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
5461 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
5521 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
5581 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
5641 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
5701 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
5761 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
5821 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
5881 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
5941 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
6001 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
6061 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
6121 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
6181 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
6241 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
6301 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
6361 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
6421 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
6481 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
6541 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
6601 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
6661 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
6721 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
6781 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
6841 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
6901 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
6961 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
7021 accgagcgca gcgagtcagt gagcgaggaa gcggaag
//
