| 质粒名称: | HA-Rab11-WT |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 101047 |
| 存储实验室: | Ken-Ichi Takemaru |
|
载体骨架基本信息 |
|
| 载体名称: | pCS2+HA |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 4100 bp |
| 完整质粒大小: | 4750 bp |
| 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: | RAB11 |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 650 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | CMV |
| 标签/融合蛋白: | HA（骨干网上的N个终端） |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | BamHI(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-Rab11-WT was a gift from Ken-Ichi Takemaru (Addgene plasmid # 101047 ; http://n2t.net/addgene:101047 ; RRID:Addgene_101047) |
| 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 |
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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-Rab11-WT 4767 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION Expresses HA-tagged human Rab11-WT in mammalian cells.
ACCESSION .
VERSION .
KEYWORDS HA-Rab11-WT
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4767)
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 4767)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..4767
/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(1773..1792)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(1774..1792)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
polyA_signal 1797..1931
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(1821..1840)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 1875..1894
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind complement(2044..2064)
/label=T3
/note="T3 promoter, forward primer"
promoter complement(2044..2062)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(2083..2099)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(2083..2099)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(2096..2118)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 2107..2123
/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(2131..2161)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 2176..2197
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(2314..2331)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(2485..3073)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(2565..2584)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(3244..4104)
/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 3867..3886
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(4105..4209)
/gene="bla"
/label=AmpR promoter
rep_origin complement(4235..4690)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(4372..4393)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 4584..4603
/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 ccggtacccg cgacgacgag tacgactacc tctttaaagt
1141 tgtccttatt ggagattctg gtgttggaaa gagtaatctc ctgtctcgat ttactcgaaa
1201 tgagtttaat ctggaaagca agagcaccat tggagtagag tttgcaacaa gaagcatcca
1261 ggttgatgga aaaacaataa aggcacagat atgggacaca gcagggcaag agcgatatcg
1321 agctataaca tcagcatatt atcgtggagc tgtaggtgcc ttattggttt atgacattgc
1381 taaacatctc acatatgaaa atgtagagcg atggctgaaa gaactgagag atcatgctga
1441 tagtaacatt gttatcatgc ttgtgggcaa taagagtgat ctacgtcatc tcagggcagt
1501 tcctacagat gaagcaagag cttttgcaga aaagaatggt ttgtcattca ttgaaacttc
1561 ggccctagac tctacaaatg tagaagctgc ttttcagaca attttaacag agatttaccg
1621 cattgtttct cagaagcaaa tgtcagacag acgcgaaaat gacatgtctc caagcaacaa
1681 tgtggttcct attcatgttc caccaaccac tgaaaacaag ccaaaggtgc agtgctgtca
1741 gaacatctaa ctcgagcctc tagattctgc agccctatag tgagtcgtat tacgtagatc
1801 cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa
1861 aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca
1921 ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag ggggaggtgt
1981 gggaggtttt ttaattcgcg gccgcggcgc caatgcattg ggcccggtac ccagcttttg
2041 ttccctttag tgagggttaa ttgcgcgctt ggcgtaatca tggtcatagc tgtttcctgt
2101 gtgaaattgt tatccgctca caattccaca caacatacga gccgggagca taaagtgtaa
2161 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
2221 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
2281 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
2341 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
2401 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
2461 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
2521 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
2581 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
2641 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
2701 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
2761 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
2821 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
2881 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
2941 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
3001 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
3061 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
3121 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
3181 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
3241 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
3301 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
3361 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
3421 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
3481 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
3541 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
3601 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
3661 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
3721 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
3781 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
3841 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
3901 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
3961 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
4021 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
4081 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
4141 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
4201 ggttccgcgc acatttcccc gaaaagtgcc acctaaattg taagcgttaa tattttgtta
4261 aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc
4321 aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg
4381 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat
4441 cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc
4501 cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag
4561 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg
4621 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta
4681 cagggcgcgt cccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg
4741 gcctcttcgc tattacgcca gtcgacc
//
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