| 质粒名称: | PDIA3-bio-His |
|---|---|
| 出品公司: | addgene |
| 目录编号: | 52070 |
| 存储实验室: | Gavin Wright |
|
载体骨架基本信息 |
|
| 载体名称: | pTT3 |
| 载体出品公司: | Yves Durocher, NRC |
| 空载体大小: | 6618 bp |
| 完整质粒大小: | 8153 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: | Top10 |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | PDIA3 / Protein disulfide-isomerase A3 precursor |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 1535 bp |
| 启动子: | CMV |
| 标签/融合蛋白: |
rat CD4 d3+4 (C terminal on backbone)
enzymatic biotinylation sequence (C terminal on backbone)
His tag (C terminal on backbone)
|
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | NotI(未破坏) |
| 3’克隆位点: | AscI (未破坏) |
| 5’测序引物 | CMV-F |
|
文献引用方法 |
|
| 材料和方法部分: | PDIA3-bio-His was a gift from Gavin Wright (Addgene plasmid # 52070 ; http://n2t.net/addgene:52070 ; RRID:Addgene_52070) |
| 参考文献部分: | A human platelet receptor protein microarray identifies FcepsilonR1alpha as an activating PEAR1 ligand. Sun Y, Vandenbriele C, Kauskot A, Verhamme P, Hoylaerts MF, Wright GJ. Mol Cell Proteomics. 2015 Feb 23. pii: mcp.M114.046946. 10.1074/mcp.M114.046946 PubMed 25713122 |
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 PDIA3-bio-His 8153 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Expresses full-length Protein disulfide-isomerase A3 precursor
ectodomain in mammalian cells. C-terminal rat Cd4d3+4 tag,
biotinylation sequence and His tag. .
ACCESSION .
VERSION .
KEYWORDS PDIA3-bio-His
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8153)
AUTHORS Sun Y, Vandenbriele C, Kauskot A, Verhamme P, Hoylaerts MF, Wright
GJ
TITLE A human platelet receptor protein microarray identifies
FcepsilonR1alpha as an activating PEAR1 ligand.
JOURNAL Mol Cell Proteomics. 2015 Feb 23. pii: mcp.M114.046946.
PUBMED 25713122
REFERENCE 2 (bases 1 to 8153)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..8153
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 1..380
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 381..584
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 535..555
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 581..605
/label=LNCX
/note="Human CMV promoter, forward primer"
primer_bind 1086..1105
/label=pMT2-F
/note="Synthetic intron, forward primer"
regulatory 1139..1148
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 3311..3328
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
polyA_signal 3380..3435
/label=-beta-globin poly(A) signal
/note="rabbit beta-globin polyadenylation signal (Gil and
Proudfoot, 1987)"
primer_bind complement(3434..3453)
/label=rbglobpA-R
/note="Rabbit beta-globin polyA, reverse primer. Also
called rb-glob-pA-term-R"
primer_bind complement(5671..5689)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 5757..5861
/gene="bla"
/label=AmpR promoter
CDS 5862..6722
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind complement(6080..6099)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 6893..7481
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 7382..7401
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 7635..7652
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 7769..7790
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 7805..7835
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 7843..7859
/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 7848..7870
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 7867..7883
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 7867..7883
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
ORIGIN
1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
241 aagtgtatca tatgccaagt ccgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
301 ggcattatgc ccagtacatg accttacggg actttcctac ttggcagtac atctacgtat
361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta caccaatggg cgtggatagc
421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaataa ccccgccccg ttgacgcaaa
541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc
601 agatcctcac tctcttccgc atcgctgtct gcgagggcca gctgttgggc tcgcggttga
661 ggacaaactc ttcgcggtct ttccagtact cttggatcgg aaacccgtcg gcctccgaac
721 ggtactccgc caccgaggga cctgagcgag tccgcatcga ccggatcgga aaacctctcg
781 agaaaggcgt ctaaccagtc acagtcgcaa ggtaggctga gcaccgtggc gggcggcagc
841 gggtggcggt cggggttgtt tctggcggag gtgctgctga tgatgtaatt aaagtaggcg
901 gtcttgagac ggcggatggt cgaggtgagg tgtggcaggc ttgagatcca gctgttgggg
961 tgagtactcc ctctcaaaag cgggcattac ttctgcgcta agattgtcag tttccaaaaa
1021 cgaggaggat ttgatattca cctggcccga tctggccata cacttgagtg acaatgacat
1081 ccactttgcc tttctctcca caggtgtcca ctcccaggtc caagtttaaa ctgcggccgc
1141 caccatggct ctgcggagac tggccctgtt tcctggcgtg gccctgctgc tggctgctgc
1201 tagactggcc gctgccagcg acgtgctgga actgaccgac gacaacttcg agagccggat
1261 cagcgatacc ggcagcgccg gactgatgct ggtcgagttc ttcgcccctt ggtgcggcca
1321 ctgcaagagg ctggcccctg agtacgaggc tgccgccacc agactgaagg gcatcgtgcc
1381 cctggccaag gtggactgca ccgccaacac caacacctgt aacaagtacg gcgtgagcgg
1441 ctaccccacc ctgaagatct tccgggacgg cgaggaagct ggcgcctacg acggccctag
1501 aaccgccgat ggcatcgtgt cccacctgaa gaagcaggcc ggaccagcca gcgtgcccct
1561 gcggaccgag gaagagttca agaagttcat cagcgacaag gacgccagca tcgtgggctt
1621 tttcgacgac agcttcagcg aggcccacag cgagtttctg aaggccgcca gcaacctgag
1681 agacaactac agattcgccc acaccaacgt ggagagcctg gtcaacgagt acgacgacaa
1741 cggcgagggc atcatcctgt tccggcccag ccacctgacc aacaagttcg aggacaagac
1801 cgtggcctac accgagcaga agatgaccag cggcaagatc aagaagttta tccaggaaaa
1861 catcttcggc atctgccccc acatgaccga ggacaacaag gacctgatcc agggcaagga
1921 tctgctgatc gcctactacg acgtggacta cgagaagaac gccaagggca gcaactactg
1981 gcggaacaga gtgatgatgg tggccaagaa gttcctggac gccggccaca agctgaactt
2041 cgccgtggcc agcagaaaga ccttcagcca cgagctgtcc gacttcggcc tggaaagcac
2101 agccggcgag atccccgtgg tggccatcag aaccgccaag ggggagaagt tcgtgatgca
2161 ggaagagttt agccgggacg gcaaggccct ggaacggttc ctgcaggact acttcgacgg
2221 caacctgaag agatacctga agtccgagcc catccccgag agcaacgacg gccccgtgaa
2281 ggtggtggtg gccgagaact tcgacgagat cgtgaacaac gagaacaagg acgtgctgat
2341 cgaattctat gctccttggt gtgggcattg caagaacctg gaacccaagt acaaagagct
2401 gggcgagaag ctgtccaagg accccaatat cgtgatcgcc aagatggacg ccacagccaa
2461 cgacgtgccc agcccttacg aagtgcgcgg cttccccacc atctacttca gccccgccaa
2521 caagaagctg aaccccaaga agtacgaggg cggcagagaa ctgagcgact tcatcagcta
2581 tctgcagcgg gaggccacca atccccctgt gatccaggaa gaaaagccca agaagaagaa
2641 aaaggcccag gaagatctcg gcgcgccgtc gacctccatc acggcctata agagtgaggg
2701 ggagtcagcg gagttctcct tcccactcaa ccttggagag gaaagcctgc agggagagtt
2761 gagatggaag gcagagaagg ctccttcttc ccagtcctgg atcaccttct ccctaaagaa
2821 ccaaaaggtg tctgtgcaga agtctactag caaccccaag ttccagctgt ccgaaacgct
2881 cccactcacc cttcagatac cccaggtctc ccttcagttt gctggttctg gcaacctgac
2941 cctgactctg gacagaggga tactgtatca ggaagtgaac ctggtggtga tgaaagtgac
3001 tcagcccgac agcaacactt tgacctgtga ggtgatggga cccacctcac ccaagatgag
3061 actgatcttg aagcaggaga atcaggaggc cagggtctcc aggcaggaga aagtgattca
3121 agtgcaggcc cctgaagcag gggtgtggca atgtctactg agtgaaggtg aagaggtcaa
3181 gatggactcc aagatccagg ttttatccaa agggttgaat tccggcagcc tgcaccacat
3241 cctggacgcc cagaagatgc tgtggaacca cagggacagg aacctgcctc ctctggcccc
3301 tctgggcccc caccaccatc accaccattg agtctagaca cgtgattaat taaggatccc
3361 ccgacctcga cctctggcta ataaaggaaa tttattttca ttgcaatagt gtgttggaat
3421 tttttgtgtc tctcactcgg aaggacatat gggagggcaa atcatttggt cgagatccct
3481 cggagatctc tagctagagg atcgatcccc gccccggacg aactaaacct gactacgaca
3541 tctctgcccc ttcttcgcgg ggcagtgcat gtaatccctt cagttggttg gtacaacttg
3601 ccaactgggc cctgttccac atgtgacacg gggggggacc aaacacaaag gggttctctg
3661 actgtagttg acatccttat aaatggatgt gcacatttgc caacactgag tggctttcat
3721 cctggagcag actttgcagt ctgtggactg caacacaaca ttgcctttat gtgtaactct
3781 tggctgaagc tcttacacca atgctggggg acatgtacct cccaggggcc caggaagact
3841 acgggaggct acaccaacgt caatcagagg ggcctgtgta gctaccgata agcggaccct
3901 caagagggca ttagcaatag tgtttataag gcccccttgt taaccctaaa cgggtagcat
3961 atgcttcccg ggtagtagta tatactatcc agactaaccc taattcaata gcatatgtta
4021 cccaacggga agcatatgct atcgaattag ggttagtaaa agggtcctaa ggaacagcga
4081 tatctcccac cccatgagct gtcacggttt tatttacatg gggtcaggat tccacgaggg
4141 tagtgaacca ttttagtcac aagggcagtg gctgaagatc aaggagcggg cagtgaactc
4201 tcctgaatct tcgcctgctt cttcattctc cttcgtttag ctaatagaat aactgctgag
4261 ttgtgaacag taaggtgtat gtgaggtgct cgaaaacaag gtttcaggtg acgcccccag
4321 aataaaattt ggacgggggg ttcagtggtg gcattgtgct atgacaccaa tataaccctc
4381 acaaacccct tgggcaataa atactagtgt aggaatgaaa cattctgaat atctttaaca
4441 atagaaatcc atggggtggg gacaagccgt aaagactgga tgtccatctc acacgaattt
4501 atggctatgg gcaacacata atcctagtgc aatatgatac tggggttatt aagatgtgtc
4561 ccaggcaggg accaagacag gtgaaccatg ttgttacact ctatttgtaa caaggggaaa
4621 gagagtggac gccgacagca gcggactcca ctggttgtct ctaacacccc cgaaaattaa
4681 acggggctcc acgccaatgg ggcccataaa caaagacaag tggccactct tttttttgaa
4741 attgtggagt gggggcacgc gtcagccccc acacgccgcc ctgcggtttt ggactgtaaa
4801 ataagggtgt aataacttgg ctgattgtaa ccccgctaac cactgcggtc aaaccacttg
4861 cccacaaaac cactaatggc accccgggga atacctgcat aagtaggtgg gcgggccaag
4921 ataggggcgc gattgctgcg atctggagga caaattacac acacttgcgc ctgagcgcca
4981 agcacagggt tgttggtcct catattcacg aggtcgctga gagcacggtg ggctaatgtt
5041 gccatgggta gcatatacta cccaaatatc tggatagcat atgctatcct aatctatatc
5101 tgggtagcat aggctatcct aatctatatc tgggtagcat atgctatcct aatctatatc
5161 tgggtagtat atgctatcct aatttatatc tgggtagcat aggctatcct aatctatatc
5221 tgggtagcat atgctatcct aatctatatc tgggtagtat atgctatcct aatctgtatc
5281 cgggtagcat atgctatcct aatagagatt agggtagtat atgctatcct aatttatatc
5341 tgggtagcat atactaccca aatatctgga tagcatatgc tatcctaatc tatatctggg
5401 tagcatatgc tatcctaatc tatatctggg tagcataggc tatcctaatc tatatctggg
5461 tagcatatgc tatcctaatc tatatctggg tagtatatgc tatcctaatt tatatctggg
5521 tagcataggc tatcctaatc tatatctggg tagcatatgc tatcctaatc tatatctggg
5581 tagtatatgc tatcctaatc tgtatccggg tagcatatgc tatcctcatg ataagctgtc
5641 aaacatgaga attttcttga agacgaaagg gcctcgtgat acgcctattt ttataggtta
5701 atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg
5761 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
5821 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
5881 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
5941 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
6001 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
6061 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtgttgac gccgggcaag
6121 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
6181 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
6241 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
6301 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
6361 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgcagca atggcaacaa
6421 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
6481 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
6541 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
6601 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
6661 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
6721 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
6781 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
6841 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
6901 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
6961 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
7021 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
7081 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
7141 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
7201 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
7261 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
7321 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
7381 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
7441 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
7501 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
7561 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
7621 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac
7681 cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact
7741 ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc
7801 aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat
7861 ttcacacagg aaacagctat gaccatgatt acgccaagct ctagctagag gtcgaccaat
7921 tctcatgttt gacagcttat catcgcagat ccgggcaacg ttgttgccat tgctgcaggc
7981 gcagaactgg taggtatgga agatctatac attgaatcaa tattggcaat tagccatatt
8041 agtcattggt tatatagcat aaatcaatat tggctattgg ccattgcata cgttgtatct
8101 atatcataat atgtacattt atattggctc atgtccaata tgaccgccat gtt
//
