| 质粒名称: | HA-Rab8a-DN (T22N) |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 101049 |
| 存储实验室: | Ken-Ichi Takemaru |
|
载体骨架基本信息 |
|
| 载体名称: | pCS2+HA |
| 空载体大小: | 4100 bp |
| 完整质粒大小: | 5843 bp |
| 载体类型: | 哺乳细胞表达 |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | Rab8a |
| 物种来源: | H. sapiens (human) |
| 基因突变: | T22N |
| 启动子: | CMV |
| 标签/融合蛋白: | HA (N terminal on backbone) |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | BamH(未破坏) |
| 3’克隆位点: | XhoI(未破坏) |
| 5’测序引物 | CCCAAGCTTGATTTAG (SP6) |
| 3’测序引物 | TTATGCTGAGTGATATC (T7) |
|
文献引用方法 |
|
| 材料和方法部分: | HA-Rab8a-DN (T22N) was a gift from Ken-Ichi Takemaru (Addgene plasmid # 101049 ; http://n2t.net/addgene:101049 ; RRID:Addgene_101049) |
| 参考文献部分: | Chibby promotes ciliary vesicle formation and basal body docking during airway cell differentiation. Burke MC, Li FQ, Cyge B, Arashiro T, Brechbuhl HM, Chen X, Siller SS, Weiss MA, O'Connell CB, Love D, Westlake CJ, Reynolds SD, Kuriyama R, Takemaru K. J Cell Biol. 2014 Oct 13;207(1):123-37. doi: 10.1083/jcb.201406140. 10.1083/jcb.201406140 PubMed 25313408 |
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 HA-Rab8a-DN (T22N) 4743 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Expresses HA-tagged human Rab8-DN (T22N) in mammalian cells.
ACCESSION .
VERSION .
KEYWORDS HA-Rab8a-DN (T22N)
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4743)
AUTHORS Burke MC, Li FQ, Cyge B, Arashiro T, Brechbuhl HM, Chen X, Siller
SS, Weiss MA, O'Connell CB, Love D, Westlake CJ, Reynolds SD,
Kuriyama R, Takemaru K
TITLE Chibby promotes ciliary vesicle formation and basal body docking
during airway cell differentiation.
JOURNAL J Cell Biol. 2014 Oct 13;207(1):123-37. doi: 10.1083/jcb.201406140.
PUBMED 25313408
REFERENCE 2 (bases 1 to 4743)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..4743
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 1..978
/label=CMV IE94 promoter
/note="enhancer/promoter region of simian cytomegalovirus
major immediate early transcription unit IE94"
promoter 1013..1031
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind 1013..1030
/label=SP6
/note="SP6 promoter, forward primer"
CDS 1070..1096
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
primer_bind complement(1749..1768)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(1750..1768)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
polyA_signal 1773..1907
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(1797..1816)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 1851..1870
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind complement(2020..2040)
/label=T3
/note="T3 promoter, forward primer"
promoter complement(2020..2038)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(2059..2075)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(2059..2075)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(2072..2094)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 2083..2099
/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(2107..2137)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 2152..2173
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(2290..2307)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(2461..3049)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(2541..2560)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(3220..4080)
/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 3843..3862
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(4081..4185)
/gene="bla"
/label=AmpR promoter
rep_origin complement(4211..4666)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(4348..4369)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 4560..4579
/label=F1ori-R
/note="F1 origin, reverse primer"
ORIGIN
1 gccaattcaa tatggcgtat atggactcat gccaattcaa tatggtggat ctggacctgt
61 gccaattcaa tatggcgtat atggactcgt gccaattcaa tatggtggat ctggacccca
121 gccaattcaa tatggcggac ttggcaccat gccaattcaa tatggcggac ctggcactgt
181 gccaactggg gaggggtcta cttggcacgg tgccaagttt gaggaggggt cttggccctg
241 tgccaagtcc gccatattga attggcatgg tgccaataat ggcggccata ttggctatat
301 gccaggatca atatataggc aatatccaat atggccctat gccaatatgg ctattggcca
361 ggttcaatac tatgtattgg ccctatgcca tatagtattc catatatggg ttttcctatt
421 gacgtagata gcccctccca atgggcggtc ccatatacca tatatggggc ttcctaatac
481 cgcccatagc cactccccca ttgacgtcaa tggtctctat atatggtctt tcctattgac
541 gtcatatggg cggtcctatt gacgtatatg gcgcctcccc cattgacgtc aattacggta
601 aatggcccgc ctggctcaat gcccattgac gtcaatagga ccacccacca ttgacgtcaa
661 tgggatggct cattgcccat tcatatccgt tctcacgccc cctattgacg tcaatgacgg
721 taaatggccc acttggcagt acatcaatat ctattaatag taacttggca agtacattac
781 tattggaagt acgccagggt acattggcag tactcccatt gacgtcaatg gcggtaaatg
841 gcccgcgatg gctgccaagt acatccccat tgacgtcaat ggggaggggc aatgacgcaa
901 atgggcgttc cattgacgta aatgggcggt aggcgtgcct aatgggaggt ctatataagc
961 aatgctcgtt tagggaaccg ccattctgcc tggggacgtc ggagcaagct tgatttaggt
1021 gacactatag aatacaagct acttgttctt tttgcaggat accatgcgat acccatatga
1081 tgtgccagat tacgctggat ccatggcgaa gacctacgat tacctgttca agctgctgct
1141 gatcggggac tcgggggtgg ggaagaactg tgtcctgttc cgcttctccg aggacgcctt
1201 caactccact tttatctcca ccataggaat tgactttaaa attaggacca tagagctcga
1261 tggcaagaga attaaactgc agatatggga cacagccggt caggaacggt ttcggacgat
1321 cacaacggcc tactacaggg gtgcaatggg catcatgctg gtctacgaca tcaccaacga
1381 gaagtccttc gacaacatcc ggaactggat tcgcaacatt gaggagcacg cctctgcaga
1441 cgtcgaaaag atgatactcg ggaacaagtg tgatgtgaat gacaagagac aagtttccaa
1501 ggaacgggga gaaaagctgg ccctcgacta tggaatcaag ttcatggaga ccagcgcgaa
1561 ggccaacatc aatgtggaaa atgcattttt cactctcgcc agagatatca aagcaaaaat
1621 ggacaaaaaa ttggaaggca acagccccca ggggagcaac cagggagtca aaatcacacc
1681 ggaccagcag aagaggagca gctttttccg atgtgttctt ctgtagctcg agcctctaga
1741 ttctgcagcc ctatagtgag tcgtattacg tagatccaga catgataaga tacattgatg
1801 agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg
1861 atgctattgc tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt
1921 gcattcattt tatgtttcag gttcaggggg aggtgtggga ggttttttaa ttcgcggccg
1981 cggcgccaat gcattgggcc cggtacccag cttttgttcc ctttagtgag ggttaattgc
2041 gcgcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
2101 tccacacaac atacgagccg ggagcataaa gtgtaaagcc tggggtgcct aatgagtgag
2161 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
2221 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
2281 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
2341 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
2401 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
2461 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
2521 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
2581 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
2641 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
2701 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
2761 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
2821 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
2881 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
2941 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
3001 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
3061 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
3121 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
3181 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
3241 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
3301 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
3361 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
3421 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
3481 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
3541 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
3601 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
3661 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
3721 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
3781 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
3841 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
3901 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
3961 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
4021 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
4081 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
4141 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
4201 agtgccacct aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
4261 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
4321 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac
4381 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa
4441 ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct
4501 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa
4561 gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc
4621 gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtccca ttcgccattc
4681 aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagtcg
4741 acc
//
