| 质粒名称: | pCI HA NEDD4 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 27002 |
| 存储实验室: | Joan Massague |
|
载体骨架基本信息 |
|
| 载体名称: | pCI |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 4000 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: | NEDD4 |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 2700 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | HA（插入N端子） |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | KpnI (未破坏) |
| 3' 克隆位点: | NotI(未破坏) |
| 5' 测序引物 | T7 |
| 3' 测序引物 | EBV-rev |
|
文献引用方法 |
|
| 材料和方法部分: | pCI HA NEDD4 was a gift from Joan Massague (Addgene plasmid # 27002 ; http://n2t.net/addgene:27002 ; RRID:Addgene_27002) |
| 参考文献部分: | Ubiquitin ligase Nedd4L targets activated Smad2/3 to limit TGF-beta signaling. Gao S, Alarcon C, Sapkota G, Rahman S, Chen PY, Goerner N, Macias MJ, Erdjument-Bromage H, Tempst P, Massague J. Mol Cell. 2009 Nov 13. 36(3):457-68. 10.1016/j.molcel.2009.09.043 PubMed 19917253 |
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 pCI HA NEDD4 6727 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pCI HA NEDD4
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6727)
AUTHORS Gao S, Alarcon C, Sapkota G, Rahman S, Chen PY, Goerner N, Macias
MJ, Erdjument-Bromage H, Tempst P, Massague J
TITLE Ubiquitin ligase Nedd4L targets activated Smad2/3 to limit TGF-beta
signaling.
JOURNAL Mol Cell. 2009 Nov 13. 36(3):457-68.
PUBMED 19917253
REFERENCE 2 (bases 1 to 6727)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6727
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 138..517
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 518..721
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 672..692
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 718..742
/label=LNCX
/note="Human CMV promoter, forward primer"
intron 857..989
/label=chimeric intron
/note="chimera between introns from human beta-globin and
immunoglobulin heavy chain genes"
primer_bind 972..991
/label=pMT2-F
/note="Synthetic intron, forward primer"
promoter 1034..1052
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 1073..1099
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
polyA_signal 3841..3962
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(3852..3871)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 3906..3925
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
rep_origin 4143..4598
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(4230..4249)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 4440..4461
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(4665..4684)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 4784..4806
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(4844..4862)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 4930..5034
/gene="bla"
/label=AmpR promoter
CDS 5035..5895
/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(5253..5272)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 6066..6654
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 6555..6574
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 tcaatattgg ccattagcca tattattcat tggttatata gcataaatca atattggcta
61 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
121 aatatgaccg ccatgttggc attgattatt gactagttat taatagtaat caattacggg
181 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
241 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
301 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
361 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
421 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
481 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
541 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
601 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
661 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
721 tcgtttagtg aaccgtcaga tcactagaag ctttattgcg gtagtttatc acagttaaat
781 tgctaacgca gtcagtgctt ctgacacaac agtctcgaac ttaagctgca gaagttggtc
841 gtgaggcact gggcaggtaa gtatcaaggt tacaagacag gtttaaggag accaatagaa
901 actgggcttg tcgagacaga gaagactctt gcgtttctga taggcaccta ttggtcttac
961 tgacatccac tttgcctttc tctccacagg tgtccactcc cagttcaatt acagctctta
1021 aggctagagt acttaatacg actcactata ggctagcctc gagaccatgg attatccgta
1081 tgatgttcct gattatgcag aattcacgcg tggtaccgca acttgcgcgg tggaggtgtt
1141 cgggctcctg gaggacgagg aaaattcacg aattgtgaga gtaagagtta tagccggaat
1201 aggccttgcc aagaaggata tattgggagc tagtgatcct tacgtgagag tgacgttata
1261 tgacccaatg aatggagttc ttacaagtgt gcaaacaaaa accattaaaa agagtttgaa
1321 tccaaagtgg aatgaagaaa tattattcag agttcatcct cagcagcacc ggcttctttt
1381 tgaagtgttt gacgaaaacc gattgacaag agatgatttc ctaggtcaag tggatgttcc
1441 actttatcca ttaccgacag aaaatccaag attggagaga ccatatacat ttaaggattt
1501 tgttcttcat ccaagaagtc acaaatcaag agttaaaggt tatctgagac taaaaatgac
1561 ttatttacct aaaaccagtg gctcagaaga tgataatgca gaacaggctg aggaattaga
1621 gcctggctgg gttgttttgg accaaccaga tgctgcttgc catttgcagc aacaacaaga
1681 accttctcct ctacctccag ggtgggaaga gaggcaggat atccttggaa ggacctatta
1741 tgtaaaccat gaatctagaa gaacacagtg gaaaagacca acccctcagg acaacctaac
1801 agatgctgag aatggcaaca ttcaactgca agcacaacgt gcatttacca ccaggcggca
1861 gatatccgag gaaacagaaa gtgttgacaa ccaagagtct tccgagaact gggaaattat
1921 aagagaagat gaagccacca tgtatagcag ccaggccttc ccatcacctc caccgtcaag
1981 taacttggat gttccaactc atcttgcaga agaattgaat gccagactca ccatttttgg
2041 aaattcagcc gtgagccagc cagcatcgag ctcaaatcat tccagcagaa gaggcagctt
2101 acaagcctat acttttgagg aacaacctac acttcctgtg cttttgccta cttcatctgg
2161 attaccacca ggttgggaag aaaaacaaga tgaaagagga agatcatatt atgtagatca
2221 caattccaga acgactactt ggacaaagcc cactgtacag gccacagtgg agaccagtca
2281 gctgacctca agccagagtt ctgcaggccc tcaatcacaa gcctccacca gtgattcagg
2341 ccagcaggtg acccagccat ctgaaattga gcaaggattc cttcctaaag gctgggaagt
2401 ccggcatgca ccaaatggga ggcctttctt tattgaccac aacactaaaa ccaccacctg
2461 ggaagatcca agattgaaaa ttccagccca tctgagagga aagacatcac ttgatacttc
2521 caatgatcta gggcctttac ctccaggatg ggaagagaga actcacacag atggaagaat
2581 cttctacata aatcacaata taaaaagaac acaatgggaa gatcctcggt tggagaatgt
2641 agcaataact ggaccagcag tgccctactc cagggattac aaaagaaagt atgagttctt
2701 ccgaagaaag ttgaagaagc agaatgacat tccaaacaaa tttgaaatga aacttcgccg
2761 agcaactgtt cttgaagact cttaccggag aattatgggt gtcaagagag cagacttcct
2821 gaaggctcga ctgtggattg agtttgatgg tgaaaaggga ttggattatg gaggagttgc
2881 cagagaatgg ttcttcctga tctcaaagga aatgtttaac ccttattatg ggttgtttga
2941 atattctgct acggacaatt ataccctaca gataaatcca aactctggat tgtgtaacga
3001 agatcacctc tcttacttca agtttattgg tcgggtagct ggaatggcag tttatcatgg
3061 caaactgttg gatggttttt tcatccgccc attttacaag atgatgcttc acaaaccaat
3121 aacccttcat gatatggaat ctgtggatag tgaatattac aattccctaa gatggattct
3181 tgaaaatgac ccaacagaat tggacctcag gtttatcata gatgaagaac tttttggaca
3241 gacacatcaa catgagctga aaaatggtgg atcagaaata gttgtcacca ataagaacaa
3301 aaaggaatat atttatcttg taatacaatg gcgatttgta aaccgaatcc agaagcaaat
3361 ggctgctttt aaagagggat tctttgaact aataccacag gatctcatca aaatttttga
3421 tgaaaatgaa ctagagcttc ttatgtgtgg actgggagat gttgatgtga atgactggag
3481 ggaacataca aagtataaaa atggctacag tgcaaatcat caggttatac agtggttttg
3541 gaaggctgtt ttaatgatgg attcagaaaa aagaataaga ttacttcagt ttgtcactgg
3601 cacatctcgg gtgcctatga atggatttgc tgaactatac ggttcaaatg gaccacagtc
3661 atttacagtt gaacagtggg gtacgcctga aaagctgcca agagctcata cctgttttaa
3721 tcgcctggac ttgccacctt atgaatcatt tgaagaatta tgggataaac ttcagatggc
3781 aattgaaaac acccagggct ttgatggagt tgattaggcg gccgcttcga gcagacatga
3841 taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta
3901 tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag
3961 ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggagatg tgggaggttt
4021 tttaaagcaa gtaaaacctc tacaaatgtg gtaaaatcga taaggatccg ggctggcgta
4081 atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat
4141 ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
4201 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
4261 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
4321 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
4381 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
4441 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
4501 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
4561 taacgcgaat tttaacaaaa tattaacgct tacaatttcc tgatgcggta ttttctcctt
4621 acgcatctgt gcggtatttc acaccgcata tggtgcactc tcagtacaat ctgctctgat
4681 gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct
4741 tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt
4801 cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta
4861 tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg
4921 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
4981 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
5041 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
5101 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
5161 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
5221 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
5281 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
5341 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
5401 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
5461 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
5521 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
5581 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
5641 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
5701 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
5761 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
5821 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
5881 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
5941 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
6001 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
6061 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
6121 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
6181 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
6241 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
6301 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
6361 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
6421 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
6481 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
6541 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
6601 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
6661 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catggctcga
6721 cagatct
//
