| 质粒名称: | CTGF-bio-His |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 52028 |
| 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: | 6618bp |
| 完整质粒大小: | 7685bp |
| 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: | CTGF / Connective tissue growth factor precursor |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 1067 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: | CTGF-bio-His was a gift from Gavin Wright (Addgene plasmid # 52028 ; http://n2t.net/addgene:52028 ; RRID:Addgene_52028) |
| 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 |
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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 CTGF-bio-His 7685 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Expresses full-length Connective tissue growth factor precursor
ectodomain in mammalian cells. C-terminal rat Cd4d3+4 tag,
biotinylation sequence and His tag..
ACCESSION .
VERSION .
KEYWORDS CTGF-bio-His
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7685)
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 7685)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..7685
/organism="synthetic DNA construct"
/mol_type="other DNA"
CDS 644..661
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
polyA_signal 713..768
/label=-beta-globin poly(A) signal
/note="rabbit beta-globin polyadenylation signal (Gil and
Proudfoot, 1987)"
primer_bind complement(767..786)
/label=rbglobpA-R
/note="Rabbit beta-globin polyA, reverse primer. Also
called rb-glob-pA-term-R"
primer_bind complement(3004..3022)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 3090..3194
/gene="bla"
/label=AmpR promoter
CDS 3195..4055
/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(3413..3432)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 4226..4814
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 4715..4734
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 4968..4985
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 5102..5123
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 5138..5168
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 5176..5192
/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 5181..5203
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 5200..5216
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 5200..5216
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
enhancer 5487..5866
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 5867..6070
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 6021..6041
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 6067..6091
/label=LNCX
/note="Human CMV promoter, forward primer"
primer_bind 6572..6591
/label=pMT2-F
/note="Synthetic intron, forward primer"
regulatory 6625..6634
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
ORIGIN
1 gtcgacctcc atcacggcct ataagagtga gggggagtca gcggagttct ccttcccact
61 caaccttgga gaggaaagcc tgcagggaga gttgagatgg aaggcagaga aggctccttc
121 ttcccagtcc tggatcacct tctccctaaa gaaccaaaag gtgtctgtgc agaagtctac
181 tagcaacccc aagttccagc tgtccgaaac gctcccactc acccttcaga taccccaggt
241 ctcccttcag tttgctggtt ctggcaacct gaccctgact ctggacagag ggatactgta
301 tcaggaagtg aacctggtgg tgatgaaagt gactcagccc gacagcaaca ctttgacctg
361 tgaggtgatg ggacccacct cacccaagat gagactgatc ttgaagcagg agaatcagga
421 ggccagggtc tccaggcagg agaaagtgat tcaagtgcag gcccctgaag caggggtgtg
481 gcaatgtcta ctgagtgaag gtgaagaggt caagatggac tccaagatcc aggttttatc
541 caaagggttg aattccggca gcctgcacca catcctggac gcccagaaga tgctgtggaa
601 ccacagggac aggaacctgc ctcctctggc ccctctgggc ccccaccacc atcaccacca
661 ttgagtctag acacgtgatt aattaaggat cccccgacct cgacctctgg ctaataaagg
721 aaatttattt tcattgcaat agtgtgttgg aattttttgt gtctctcact cggaaggaca
781 tatgggaggg caaatcattt ggtcgagatc cctcggagat ctctagctag aggatcgatc
841 cccgccccgg acgaactaaa cctgactacg acatctctgc cccttcttcg cggggcagtg
901 catgtaatcc cttcagttgg ttggtacaac ttgccaactg ggccctgttc cacatgtgac
961 acgggggggg accaaacaca aaggggttct ctgactgtag ttgacatcct tataaatgga
1021 tgtgcacatt tgccaacact gagtggcttt catcctggag cagactttgc agtctgtgga
1081 ctgcaacaca acattgcctt tatgtgtaac tcttggctga agctcttaca ccaatgctgg
1141 gggacatgta cctcccaggg gcccaggaag actacgggag gctacaccaa cgtcaatcag
1201 aggggcctgt gtagctaccg ataagcggac cctcaagagg gcattagcaa tagtgtttat
1261 aaggccccct tgttaaccct aaacgggtag catatgcttc ccgggtagta gtatatacta
1321 tccagactaa ccctaattca atagcatatg ttacccaacg ggaagcatat gctatcgaat
1381 tagggttagt aaaagggtcc taaggaacag cgatatctcc caccccatga gctgtcacgg
1441 ttttatttac atggggtcag gattccacga gggtagtgaa ccattttagt cacaagggca
1501 gtggctgaag atcaaggagc gggcagtgaa ctctcctgaa tcttcgcctg cttcttcatt
1561 ctccttcgtt tagctaatag aataactgct gagttgtgaa cagtaaggtg tatgtgaggt
1621 gctcgaaaac aaggtttcag gtgacgcccc cagaataaaa tttggacggg gggttcagtg
1681 gtggcattgt gctatgacac caatataacc ctcacaaacc ccttgggcaa taaatactag
1741 tgtaggaatg aaacattctg aatatcttta acaatagaaa tccatggggt ggggacaagc
1801 cgtaaagact ggatgtccat ctcacacgaa tttatggcta tgggcaacac ataatcctag
1861 tgcaatatga tactggggtt attaagatgt gtcccaggca gggaccaaga caggtgaacc
1921 atgttgttac actctatttg taacaagggg aaagagagtg gacgccgaca gcagcggact
1981 ccactggttg tctctaacac ccccgaaaat taaacggggc tccacgccaa tggggcccat
2041 aaacaaagac aagtggccac tctttttttt gaaattgtgg agtgggggca cgcgtcagcc
2101 cccacacgcc gccctgcggt tttggactgt aaaataaggg tgtaataact tggctgattg
2161 taaccccgct aaccactgcg gtcaaaccac ttgcccacaa aaccactaat ggcaccccgg
2221 ggaatacctg cataagtagg tgggcgggcc aagatagggg cgcgattgct gcgatctgga
2281 ggacaaatta cacacacttg cgcctgagcg ccaagcacag ggttgttggt cctcatattc
2341 acgaggtcgc tgagagcacg gtgggctaat gttgccatgg gtagcatata ctacccaaat
2401 atctggatag catatgctat cctaatctat atctgggtag cataggctat cctaatctat
2461 atctgggtag catatgctat cctaatctat atctgggtag tatatgctat cctaatttat
2521 atctgggtag cataggctat cctaatctat atctgggtag catatgctat cctaatctat
2581 atctgggtag tatatgctat cctaatctgt atccgggtag catatgctat cctaatagag
2641 attagggtag tatatgctat cctaatttat atctgggtag catatactac ccaaatatct
2701 ggatagcata tgctatccta atctatatct gggtagcata tgctatccta atctatatct
2761 gggtagcata ggctatccta atctatatct gggtagcata tgctatccta atctatatct
2821 gggtagtata tgctatccta atttatatct gggtagcata ggctatccta atctatatct
2881 gggtagcata tgctatccta atctatatct gggtagtata tgctatccta atctgtatcc
2941 gggtagcata tgctatcctc atgataagct gtcaaacatg agaattttct tgaagacgaa
3001 agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga
3061 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
3121 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
3181 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
3241 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
3301 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
3361 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
3421 gcgcggtatt atcccgtgtt gacgccgggc aagagcaact cggtcgccgc atacactatt
3481 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
3541 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
3601 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
3661 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
3721 gtgacaccac gatgcctgca gcaatggcaa caacgttgcg caaactatta actggcgaac
3781 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
3841 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
3901 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
3961 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
4021 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
4081 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
4141 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
4201 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
4261 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
4321 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
4381 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
4441 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
4501 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
4561 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
4621 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
4681 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
4741 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
4801 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
4861 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
4921 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
4981 gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc
5041 attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa
5101 ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc
5161 gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg
5221 attacgccaa gctctagcta gaggtcgacc aattctcatg tttgacagct tatcatcgca
5281 gatccgggca acgttgttgc cattgctgca ggcgcagaac tggtaggtat ggaagatcta
5341 tacattgaat caatattggc aattagccat attagtcatt ggttatatag cataaatcaa
5401 tattggctat tggccattgc atacgttgta tctatatcat aatatgtaca tttatattgg
5461 ctcatgtcca atatgaccgc catgttgaca ttgattattg actagttatt aatagtaatc
5521 aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt
5581 aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta
5641 tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg
5701 gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtccgc cccctattga
5761 cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct tacgggactt
5821 tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg
5881 gcagtacacc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc
5941 cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg
6001 taataacccc gccccgttga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat
6061 aagcagagct cgtttagtga accgtcagat cctcactctc ttccgcatcg ctgtctgcga
6121 gggccagctg ttgggctcgc ggttgaggac aaactcttcg cggtctttcc agtactcttg
6181 gatcggaaac ccgtcggcct ccgaacggta ctccgccacc gagggacctg agcgagtccg
6241 catcgaccgg atcggaaaac ctctcgagaa aggcgtctaa ccagtcacag tcgcaaggta
6301 ggctgagcac cgtggcgggc ggcagcgggt ggcggtcggg gttgtttctg gcggaggtgc
6361 tgctgatgat gtaattaaag taggcggtct tgagacggcg gatggtcgag gtgaggtgtg
6421 gcaggcttga gatccagctg ttggggtgag tactccctct caaaagcggg cattacttct
6481 gcgctaagat tgtcagtttc caaaaacgag gaggatttga tattcacctg gcccgatctg
6541 gccatacact tgagtgacaa tgacatccac tttgcctttc tctccacagg tgtccactcc
6601 caggtccaag tttaaactgc ggccgccacc atggtcgccg cttctatggg acctgtgcgg
6661 gtggcctttg tggtgctgct ggccctgtgt agcagacccg ccgtgggcca gaattgcagc
6721 ggcccctgca gatgtcctga cgagcctgcc cctagatgtc ctgccggcgt gagcctggtg
6781 ctggacggct gcggctgctg cagagtgtgt gccaagcagc tgggcgagct gtgcaccgag
6841 agagatccct gcgaccccca caagggcctg ttctgccact tcggcagccc cgccaacaga
6901 aagatcggcg tctgcaccgc taaagatggc gccccttgca tcttcggcgg caccgtgtac
6961 cgatctggag agagcttcca gagcagctgc aagtaccagt gcacctgtct ggatggcgct
7021 gtcggctgca tgcccctgtg cagcatggat gtgcggctgc ccagccccga ctgccccttc
7081 cccagaagag tgaagctgcc cggcaagtgc tgcgaggaat gggtctgcga cgagcccaag
7141 gatcagacag tcgtgggacc cgctctggcc gcctaccggc tggaagatac cttcggcccc
7201 gaccccacca tgatccgggc caactgcctg gtgcagacca ccgagtggag cgcctgcagc
7261 aagacctgtg gcatgggcat cagcaccaga gtgaccaacg acaacgccag ctgccggctg
7321 gaaaagcaga gccggctgtg catggtccgg ccttgcgaag ctgacctgga agagaacatc
7381 aagaaaggca agaagtgcat ccggaccccc aagatcagca agcccatcaa gttcgaactc
7441 tctggatgca ccagcatgaa aacctaccgg gccaagttct gcggagtgtg cactgatggc
7501 agatgctgca ccccccacag aaccaccacc ctgcccgtgg agttcaagtg ccccgacggc
7561 gaagtgatga agaaaaacat gatgttcatc aagacatgcg cctgccacta caactgccct
7621 ggcgacaacg acatcttcga gagcctgtac taccggaaga tgtatggaga tatggcaggc
7681 gcgcc
//
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