| 出品公司: | ZYbscience |
|---|---|
| 载体名称: | pTYB11 |
| 质粒类型: | 原核细胞表达载体 |
| 高拷贝/低拷贝: | -- |
| 启动子: | -- |
| 克隆方法: | 多克隆位点,限制性内切酶 |
| 载体大小: | 7412bp |
| 5' 测序引物及序列: | T7 |
| 3' 测序引物及序列: | -- |
| 载体标签: | Intein |
| 载体抗性: | 氨苄青霉素(Ampicillin) |
| 筛选标记: | -- |
| 备注: | -- |
| 产品目录号: | ZY6901S |
| 稳定性: | -- |
| 组成型: | -- |
| 病毒/非病毒: | -- |
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- 详细信息
- 文献和实验
- 技术资料
- 保存条件:
-20℃低温保存
- 保质期:
三年
- 英文名:
pTYB11
- 库存:
20
- 供应商:
泽叶生物
载体基本信息
载体质粒图谱和多克隆位点信息
载体简介
pTYB11 is an E. coli cloning and expression vector (7414 bp) used in the IMPACT Protein Purification System which allows the overexpression of a target protein as a fusion to a self-cleavable affinity tag. It is a N-terminal fusion vector designed for in-frame insertion of a target gene into the polylinker, downstream of the intein tag (the Sce VMA intein/chitin binding domain, 55 kDa). This allows the N-terminus of the target protein to be fused to the intein tag. The self-cleavage activity of the intein allows the release of the target protein from the chitin-bound intein tag, resulting in a single column purification of the target protein. This vector can be used for expression and purification of a native target protein without any vector-derived residues.
载体序列
LOCUS pTYB11 7412 bp DNA SYN
DEFINITION pTYB11
ACCESSION
KEYWORDS
SOURCE
ORGANISM other sequences; artificial sequences; vectors.
FEATURES Location/Qualifiers
source 1..7412
/organism="pTYB11"
/mol_type="other DNA"
terminator 1..28
/label="rrnB_T2_terminator"
promoter 70..98
/label="AmpR_promoter"
gene 140..1000
/label="Ampicillin"
/gene="Ampicillin"
CDS 140..1000
/label="ORF frame 2"
rep_origin complement(1232..1538)
/label="f1_origin"
rep_origin 1650..2269
/label="pBR322_origin"
misc_feature 2607..2629
/label="pGEX_3_primer"
misc_feature complement(2623..2814)
/label="ROP"
CDS 2953..3588
/label="ORF frame 1"
gene complement(3010..3309)
/label="tet (611 - 312)"
/gene="tet (611 - 312)"
misc_feature complement(3371..4462)
/label="lacI"
/label="ORF frame 2"
CDS complement(4203..4703)
/label="ORF frame 1"
gene complement(4554..4805)
/label="tet (887 - 636)"
/gene="tet (887 - 636)"
misc_feature 4737..4756
/label="pBRrevBam_primer"
promoter 4840..4858
/label="T7_promoter"
misc_feature 4858..4885
/label="lacO"
misc_feature 4903..4919
/label="T7_transl_en_RBS"
CDS 4928..6559
/label="ORF frame 2"
terminator 6579..6707
/label="T7_terminator"
misc_feature complement(6621..6639)
/label="T7_Terminal_primer"
promoter complement(6870..6886)
/label="M13_forward20_primer"
misc_feature 6879..7022
/label="lacZ_a"
misc_feature complement(6879..6901)
/label="M13_pUC_fwd_primer"
misc_feature complement(7065..7082)
/label="pTrcHis_rev_primer"
misc_feature complement(7065..7082)
/label="pBAD_rev_primer"
terminator 7115..7272
/label="rrnB_terminator"
terminator 7238..7281
/label="rrnB_T1_terminator"
ORIGIN
1 AGAAGGCCAT CCTGACGGAT GGCCTTTTTG CGTTTCTACA AACTCTTTTT GTTTATTTTT
61 CTAAATACAT TCAAATATGT ATCCGCTCAT GAGACAATAA CCCTGATAAA TGCTTCAATA
121 ATATTGAAAA AGGAAGAGTA TGAGTATTCA ACATTTCCGT GTCGCCCTTA TTCCCTTTTT
181 TGCGGCATTT TGCCTTCCTG TTTTTGCTCA CCCAGAAACG CTGGTGAAAG TAAAAGATGC
241 TGAAGATCAG TTGGGTGCAC GAGTGGGTTA CATCGAACTG GATCTCAACA GCGGTAAGAT
301 CCTTGAGAGT TTTCGCCCCG AAGAACGTTC TCCAATGATG AGCACTTTTA AAGTTCTGCT
361 ATGTGGCGCG GTATTATCCC GTGTTGACGC CGGGCAAGAG CAACTCGGTC GCCGCATACA
421 CTATTCTCAG AATGACTTGG TTGAGTACTC ACCAGTCACA GAAAAGCATC TTACGGATGG
481 CATGACAGTA AGAGAATTAT GCAGTGCTGC CATAACCATG AGTGATAACA CTGCGGCCAA
541 CTTACTTCTG ACAACGATCG GAGGACCGAA GGAGCTAACC GCTTTTTTGC ACAACATGGG
601 GGATCATGTA ACTCGCCTTG ATCGTTGGGA ACCGGAGCTG AATGAAGCCA TACCAAACGA
661 CGAGCGTGAC ACCACGATGC CTGTAGCAAT GGCAACAACG TTGCGCAAAC TATTAACTGG
721 CGAACTACTT ACTCTAGCTT CCCGGCAACA ATTAATAGAC TGGATGGAGG CGGATAAAGT
781 TGCAGGACCA CTTCTGCGCT CGGCCCTTCC GGCTGGCTGG TTTATTGCTG ATAAATCTGG
841 AGCCGGTGAG CGTGGGTCTC GCGGTATCAT TGCAGCACTG GGGCCAGATG GTAAGCCCTC
901 CCGTATCGTA GTTATCTACA CGACGGGGAG TCAGGCAACT ATGGATGAAC GAAATAGACA
961 GATCGCTGAG ATAGGTGCCT CACTGATTAA GCATTGGTAA CTGTCAGACC AAGTTTACTC
1021 ATATATACTT TAGATTGATT TACCCCGGTT GATAATCAGA AAAGCCCCAA AAACAGGAAG
1081 ATTGTATAAG CAAATATTTA AATTGTAAAC GTTAATATTT TGTTAAAATT CGCGTTAAAT
1141 TTTTGTTAAA TCAGCTCATT TTTTAACCAA TAGGCCGAAA TCGGCAAAAT CCCTTATAAA
1201 TCAAAAGAAT AGCCCGAGAT AGGGTTGAGT GTTGTTCCAG TTTGGAACAA GAGTCCACTA
1261 TTAAAGAACG TGGACTCCAA CGTCAAAGGG CGAAAAACCG TCTATCAGGG CGATGGCCCA
1321 CTACGTGAAC CATCACCCAA ATCAAGTTTT TTGGGGTCGA GGTGCCGTAA AGCACTAAAT
1381 CGGAACCCTA AAGGGAGCCC CCGATTTAGA GCTTGACGGG GAAAGCCGGC GAACGTGGCG
1441 AGAAAGGAAG GGAAGAAAGC GAAAGGAGCG GGCGCTAGGG CGCTGGCAAG TGTAGCGGTC
1501 ACGCTGCGCG TAACCACCAC ACCCGCCGCG CTTAATGCGC CGCTACAGGG CGCGTAAAAG
1561 GATCTAGGTG AAGATCCTTT TTGATAATCT CATGACCAAA ATCCCTTAAC GTGAGTTTTC
1621 GTTCCACTGA GCGTCAGACC CCGTAGAAAA GATCAAAGGA TCTTCTTGAG ATCCTTTTTT
1681 TCTGCGCGTA ATCTGCTGCT TGCAAACAAA AAAACCACCG CTACCAGCGG TGGTTTGTTT
1741 GCCGGATCAA GAGCTACCAA CTCTTTTTCC GAAGGTAACT GGCTTCAGCA GAGCGCAGAT
1801 ACCAAATACT GTCCTTCTAG TGTAGCCGTA GTTAGGCCAC CACTTCAAGA ACTCTGTAGC
1861 ACCGCCTACA TACCTCGCTC TGCTAATCCT GTTACCAGTG GCTGCTGCCA GTGGCGATAA
1921 GTCGTGTCTT ACCGGGTTGG ACTCAAGACG ATAGTTACCG GATAAGGCGC AGCGGTCGGG
1981 CTGAACGGGG GGTTCGTGCA CACAGCCCAG CTTGGAGCGA ACGACCTACA CCGAACTGAG
2041 ATACCTACAG CGTGAGCTAT GAGAAAGCGC CACGCTTCCC GAAGGGAGAA AGGCGGACAG
2101 GTATCCGGTA AGCGGCAGGG TCGGAACAGG AGAGCGCACG AGGGAGCTTC CAGGGGGAAA
2161 CGCCTGGTAT CTTTATAGTC CTGTCGGGTT TCGCCACCTC TGACTTGAGC GTCGATTTTT
2221 GTGATGCTCG TCAGGGGGGC GGAGCCTATG GAAAAACGCC AGCAACGCGG CCTTTTTACG
2281 GTTCCTGGCC TTTTGCTGGC CTTTTGCTCA CATGTTCTTT CCTGCGTTAT CCCCTGATTC
2341 TGTGGATAAC CGTATTACCG CCTTTGAGTG AGCTGATACC GCTCGCCGCA GCCGAACGAC
2401 CGAGCGCAGC GAGTCAGTGA GCGAGGAAGC TATGGTGCAC TCTCAGTACA ATCTGCTCTG
2461 ATGCCGCATA GTTAAGCCAG TATACACTCC GCTATCGCTA CGTGACTGGG TCATGGCTGC
2521 GCCCCGACAC CCGCCAACAC CCGCTGACGC GCCCTGACGG GCTTGTCTGC TCCCGGCATC
2581 CGCTTACAGA CAAGCTGTGA CCGTCTCCGG GAGCTGCATG TGTCAGAGGT TTTCACCGTC
2641 ATCACCGAAA CGCGCGAGGC AGCTGCGGTA AAGCTCATCA GCGTGGTCGT GCAGCGATTC
2701 ACAGATGTCT GCCTGTTCAT CCGCGTCCAG CTCGTTGAGT TTCTCCAGAA GCGTTAATGT
2761 CTGGCTTCTG ATAAAGCGGG CCATGTTAAG GGCGGTTTTT TCCTGTTTGG TCACTGATGC
2821 CTCCGTGTAA GGGGGATTTC TGTTCATGGG GGTAATGATA CCGATGAAAC GAGAGAGGAT
2881 GCTCACGATA CGGGTTACTG ATGATGAACA TGCCCGGTTA CTGGAACGTT GTGAGGGTAA
2941 ACAACTGGCG GTATGGATGC GGCGGGACCA GAGAAAAATC ACTCAGGGTC AATGCCAGCC
3001 GAACGCCAGC AAGACGTAGC CCAGCGCGTC GGCCGCCATG CCGGCGATAA TGGCCTGCTT
3061 CTCGCCGAAA CGTTTGGTGG CGGGACCAGT GACGAAGGCT TGAGCGAGGG CGTGCAAGAT
3121 TCCGAATACC GCAAGCGACA GGCCGATCAT CGTCGCGCTC CAGCGAAAGC GGTCCTCGCC
3181 GAAAATGACC CAGAGCGCTG CCGGCACCTG TCCTACGAGT TGCATGATAA AGAAGACAGT
3241 CATAAGTGCG GCGACGATAG TCATGCCCCG CGCCCACCGG AAGGAGCTGA CTGGGTTGAA
3301 GGCTCTCAAG GGCATCGGTC GAGATCCCGG TGCCTAATGA GTGAGCTAAC TTACATTAAT
3361 TGCGTTGCGC TCACTGCCCG CTTTCCAGTC GGGAAACCTG TCGTGCCAGC TGCATTAATG
3421 AATCGGCCAA CGCGCGGGGA GAGGCGGTTT GCGTATTGGG CGCCAGGGTG GTTTTTCTTT
3481 TCACCAGTGA GACGGGCAAC AGCTGATTGC CCTTCACCGC CTGGCCCTGA GAGAGTTGCA
3541 GCAAGCGGTC CACGCTGGTT TGCCCCAGCA GGCGAAAATC CTGTTTGATG GTGGTTAACG
3601 GCGGGATATA ACATGAGCTG TCTTCGGTAT CGTCGTATCC CACTACCGAG ATATCCGCAC
3661 CAACGCGCAG CCCGGACTCG GTAATGGCGC GCATTGCGCC CAGCGCCATC TGATCGTTGG
3721 CAACCAGCAT CGCAGTGGGA ACGATGCCCT CATTCAGCAT TTGCATGGTT TGTTGAAAAC
3781 CGGACATGGC ACTCCAGTCG CCTTCCCGTT CCGCTATCGG CTGAATTTGA TTGCGAGTGA
3841 GATATTTATG CCAGCCAGCC AGACGCAGAC GCGCCGAGAC AGAACTTAAT GGGCCCGCTA
3901 ACAGCGCGAT TTGCTGGTGA CCCAATGCGA CCAGATGCTC CACGCCCAGT CGCGTACCGT
3961 CTTCATGGGA GAAAATAATA CTGTTGATGG GTGTCTGGTC AGAGACATCA AGAAATAACG
4021 CCGGAACATT AGTGCAGGCA GCTTCCACAG CAATGGCATC CTGGTCATCC AGCGGATAGT
4081 TAATGATCAG CCCACTGACG CGTTGCGCGA GAAGATTGTG CACCGCCGCT TTACAGGCTT
4141 CGACGCCGCT TCGTTCTACC ATCGACACCA CCACGCTGGC ACCCAGTTGA TCGGCGCGAG
4201 ATTTAATCGC CGCGACAATT TGCGACGGCG CGTGCAGGGC CAGACTGGAG GTGGCAACGC
4261 CAATCAGCAA CGACTGTTTG CCCGCCAGTT GTTGTGCCAC GCGGTTGGGA ATGTAATTCA
4321 GCTCCGCCAT CGCCGCTTCC ACTTTTTCCC GCGTTTTCGC AGAAACGTGG CTGGCCTGGT
4381 TCACCACGCG GGAAACGGTC TGATAAGAGA CACCGGCATA CTCTGCGACA TCGTATAACG
4441 TTACTGGTTT CACATTCACC ACCCTGAATT GACTCTCTTC CGGGCGCTAT CATGCCATAC
4501 CGCGAAAGGT TTTGCGCCAT TCGATGGTGT CCGGGATCTC GACGCTCTCC CTTATGCGAC
4561 TCCTGCATTA GGAAGCAGCC CAGTAGTAGG TTGAGGCCGT TGAGCACCGC CGCCGCAAGG
4621 AATGGTGCAT GCAAGGAGAT GGCGCCCAAC AGTCCCCCGG CCACGGGGCC TGCCACCATA
4681 CCCACGCCGA AACAAGCGCT CATGAGCCCG AAGTGGCGAG CCCGATCTTC CCCATCGGTG
4741 ATGTCGGCGA TATAGGCGCC AGCAACCGCA CCTGTGGCGC CGGTGATGCC GGCCACGATG
4801 CGTCCGGCGT AGAGGATCGA GATCTCGATC CCGCGAAATT AATACGACTC ACTATAGGGG
4861 AATTGTGAGC GGATAACAAT TCCCCTCTAG AAATAATTTT GTTTAACTTT AAGAAGGAGA
4921 TATACTTATG AAAATCGAAG AAGGTAAACT GGTAATCGGT TCCCTGGAGG GTTGCTTTGC
4981 CAAGGGTACC AATGTTTTAA TGGCGGATGG GTCTATTGAA TGTATTGAAA ACATTGAGGT
5041 TGGTAATAAG GTCATGGGTA AAGATGGCAG ACCTCGTGAG GTAATTAAAT TGCCCAGAGG
5101 AAGAGAAACT ATGTACAGCG TCGTGCAGAA AAGTCAGCAC AGAGCCCACA AAAGTGACTC
5161 AAGTCGTGAA GTGCCAGAAT TACTCAAGTT TACGTGTAAT GCGACCCATG AGTTGGTTGT
5221 TAGAACACCT CGTAGTGTCC GCCGTTTGTC TCGTACCATT AAGGGTGTCG AATATTTTGA
5281 AGTTATTACT TTTGAGATGG GCCAAAAGAA AGCCCCCGAC GGTAGAATTG TTGAGCTTGT
5341 CAAGGAAGTT TCAAAGAGCT ACCCAATATC TGAGGGGCCT GAGAGAGCCA ACGAATTAGT
5401 AGAATCCTAT AGAAAGGCTT CAAATAAAGC TTATTTTGAG TGGACTATTG AGGCCAGAGA
5461 TCTTTCTCTG TTGGGTTCCC ATGTTCGTAA AGCTACCTAC CAGACTTACG CTCCAATTCT
5521 TTATGAGAAT GACCACTTTT TCGACTACAT GCAAAAAAGT AAGTTTCATC TCACCATTGA
5581 AGGTCCAAAA GTACTTGCTT ATTTACTTGG TTTATGGATT GGTGATGGAT TGTCTGACAG
5641 GGCAACTTTT TCGGTTGATT CCAGAGATAC TTCTTTGATG GAACGTGTTA CTGAATATGC
5701 TGAAAAGTTG AATTTGTGCG CCGAGTATAA GGACAGAAAA GAACCACAAG TTGCCAAAAC
5761 TGTTAATTTG TACTCTAAAG TTGTCAGAGG TGCTAGCACA AATCCTGGTG TATCCGCTTG
5821 GCAGGTCAAC ACAGCTTATA CTGCGGGACA ATTGGTCACA TATAACGGCA AGACGTATAA
5881 ATGTTTGCAG CCCCACACCT CCTTGGCAGG ATGGGAACCA TCCAACGTTC CTGCCTTGTG
5941 GCAGCTTCAA GGTGGCCATG GTGGTATTCG CAATAATCTT AATACTGAGA ATCCATTATG
6001 GGACGCTATT GTTGGCTTAG GATTCTTGAA GGACGGTGTC AAAAATATTC CTTCTTTCTT
6061 GTCTACGGAC AATATCGGTA CTCGTGAAAC ATTTCTTGCT GGTCTAATTG ATTCTGATGG
6121 CTATGTTACT GATGAGCATG GTATTAAAGC AACAATAAAG ACAATTCATA CTTCTGTCAG
6181 AGATGGTTTG GTTTCCCTTG CTCGTTCTTT AGGCTTAGTA GTCTCGGTTA ACGCAGAACC
6241 TGCTAAGGTT GACATGAATG TCACCAAACA TAAAATTAGT TATGCTATTT ATATGTCTGG
6301 TGGAGATGTT TTGCTTAACG TTCTTTCGAA GTGTGCCGGC TCTAAAAAAT TCAGGCCTGC
6361 TCCCGCCGCT GCTTTTGCAC GTGAGTGCCG CGGATTTTAT TTCGAGTTAC AAGAATTGAA
6421 GGAAGACGAT TATTATGGGA TTACTTTATC TGATGATTCT GATCATCAGT TTTTGCTTGG
6481 ATCCCAGGTT GTTGTACAGA ACAGAAGAGC TACTAGTTCG CGAGTCGACG GCGGCCGCGA
6541 ATTCCTCGAG CCCGGGTGAC TGCAGGAAGG GGATCCGGCT GCTAACAAAG CCCGAAAGGA
6601 AGCTGAGTTG GCTGCTGCCA CCGCTGAGCA ATAACTAGCA TAACCCCTTG GGGCCTCTAA
6661 ACGGGTCTTG AGGGGTTTTT TGCTGAAAGG AGGAACTATA TCCGGATATC CCGCAAGAGG
6721 CCCGGCAGTA CCGGCATAAC CAAGCCTATG CCTACAGCAT CCAGGGTGAC GGTGCCGAGG
6781 ATGACGATGA GCGCATTGTT AGATTTCATA CACGGTGCCT GACTGCGTTA GCAATTTAAC
6841 TGTGATAAAC TACCGCATTA AAGCTTGGCA CTGGCCGTCG TTTTACAACG TCGTGACTGG
6901 GAAAACCCTG GCGTTACCCA ACTTAATCGC CTTGCAGCAC ATCCCCCTTT CGCCAGCTGG
6961 CGTAATAGCG AAGAGGCCCG CACCGATCGC CCTTCCCAAC AGTTGCGCAG CCTGAATGGC
7021 GAATGGCAGC TTGGCTGTTT TGGCGGATGA GATAAGATTT TCAGCCTGAT ACAGATTAAA
7081 TCAGAACGCA GAAGCGGTCT GATAAAACAG AATTTGCCTG GCGGCAGTAG CGCGGTGGTC
7141 CCACCTGACC CCATGCCGAA CTCAGAAGTG AAACGCCGTA GCGCCGATGG TAGTGTGGGG
7201 TCTCCCCATG CGAGAGTAGG GAACTGCCAG GCATCAAATA AAACGAAAGG CTCAGTCGAA
7261 AGACTGGGCC TTTCGTTTTA TCTGTTGTTT GTCGGTGAAC GCTCTCCTGA GTAGGACAAA
7321 TCCGCCGGGA GCGGATTTGA ACGTTGCGAA GCAACGGCCC GGAGGGTGGC GGGCAGGACG
7381 CCCGCCATAA ACTGCCAGGC ATCAAATTAA GC
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文献和实验【讲座】11月30日8-10PM 病毒基因工程国家重点实验室 吴小兵博士 病毒载体与基因治疗系列讲座
【申请置顶】 【讲座预告】11月30日 病毒基因工程国家重点实验室 吴小兵博士 病毒载体与基因治疗系列讲座 专家讲座活动通知表: 主讲人姓名:吴小兵 ========================================== 主讲人个人成就简介 吴小兵博士做为课题负责人承担了863计划基因治疗重大关键项目课题“应用于恶性肿瘤基因治疗的新型病毒载体的研制”(2000年结题)和攀登计划课题子课题“用于基因治疗的新型载体的研制”(1999年结题), 863
-CN蛋白纯化 系统,可利用还原剂DTT使标签自动裂解,无需外加蛋白酶切割,可以在4度条件下进行,小分子的还原剂也很容易去除;IMPACT-CN蛋白纯化系统包含的表达载体(pTYB载体)有两种,内含肽分别与目的蛋白的C末端(pTYB1和 pTYB2)或N末端(pTYB11和pTYB12)融合.这种在融合蛋白构建方面的机动性极大优化了目的蛋白成功表达和纯化的能力.pTYB2和 pTYB12载体的多克隆位点区设计了相同或者可兼容的酶切位点,使得目的蛋白可选择任一种融合构建载体.而pTYB1及pTYB
(pTYB载体)有两种,内含肽分别与目的蛋白的C末端(pTYB1和pTYB2)或N末端(pTYB11和pTYB12)融合。这种在融合蛋白构建方面的机动性极大优化了目的蛋白成功表达和纯化的能力。pTYB2和pTYB12载体的多克隆位点区设计了相同或者可兼容的酶切位点,使得目的蛋白可选择任一种融合构建载体。而pTYB1及pTYB11载体可让目的基因与内含肽的裂解位点直接相连,在4℃利用 DTT 诱导内含肽的肽键裂解活性,使得最后纯化得到的天然蛋白不带有源自于载体的额外的蛋白残基,同时低温也最大程度地保
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