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- 详细信息
- 文献和实验
- 技术资料
- 保存条件:
-20℃
- 保质期:
6个月
- 英文名:
pET-5a(pET5a)(60908-2772)
- 库存:
大量
- 供应商:
联迈生物
- 规格:
1ug
质粒名称: pET5a
其他名称: pET-5a
质粒大小(bp): 4134
抗性: Amp
质粒类型: 大肠杆菌表达载体
是否病毒: 非病毒
启动子:
稳转或瞬转: 瞬时表达
组成型或诱导性: 诱导表达
表达水平:
蛋白标签:
测序引物:
GenBank号:
其他属性: Hosts: E.coli. Related vectors: pBR322.
质粒图谱:
| Feature Name | Start | End | |
|---|---|---|---|
| T7_leader | 45 | 13 | |
| Xpress_fwd_primer | 46 | 28 | |
| T7_transl_en_RBS | 70 | 54 | |
| T7_promoter | 125 | 107 | |
| tet (300 – 1190) | 157 | 1047 | |
| pBRrevBam_primer | 228 | 209 | |
| ROP | 1686 | 1877 | |
| pGEX_3_primer | 1893 | 1871 | |
| Ampicillin | 3926 | 3066 | |
| AmpR_promoter | 3996 | 3968 |
| ORF | Start | End | |
|---|---|---|---|
| ORF frame 1 | 262 | 1047 | |
| ORF frame 1 | 852 | 394 | |
| ORF frame 2 | 3926 | 3066 |
| Enzyme Name | Cut |
|---|---|
| BamHI | 4 |
| NheI | 37 |
| NdeI | 44 |
| XbaI | 82 |
| BglII | 140 |
| SalI | 422 |
| EagI | 710 |
| NruI | 745 |
| MscI | 1217 |
| PvuII | 1837 |
| PstI | 3384 |
| AatII | 4061 |
| EcoRI | 4132 |
质粒序列:
| 1 | TTCGGATCCG | CGACCCATTT | GCTGTCCACC | AGTCATGCTA | GCCATATGTA | 50 |
| 51 | TATCTCCTTC | TTAAAGTTAA | ACAAAATTAT | TTCTAGAGGG | AAACCGTTGT | 100 |
| 101 | GGTCTCCCTA | TAGTGAGTCG | TATTAATTTC | GCGGGATCGA | GATCTCGATC | 150 |
| 151 | CTCTACGCCG | GACGCATCGT | GGCCGGCATC | ACCGGCGCCA | CAGGTGCGGT | 200 |
| 201 | TGCTGGCGCC | TATATCGCCG | ACATCACCGA | TGGGGAAGAT | CGGGCTCGCC | 250 |
| 251 | ACTTCGGGCT | CATGAGCGCT | TGTTTCGGCG | TGGGTATGGT | GGCAGGCCCC | 300 |
| 301 | GTGGCCGGGG | GACTGTTGGG | CGCCATCTCC | TTGCATGCAC | CATTCCTTGC | 350 |
| 351 | GGCGGCGGTG | CTCAACGGCC | TCAACCTACT | ACTGGGCTGC | TTCCTAATGC | 400 |
| 401 | AGGAGTCGCA | TAAGGGAGAG | CGTCGACCGA | TGCCCTTGAG | AGCCTTCAAC | 450 |
| 451 | CCAGTCAGCT | CCTTCCGGTG | GGCGCGGGGC | ATGACTATCG | TCGCCGCACT | 500 |
| 501 | TATGACTGTC | TTCTTTATCA | TGCAACTCGT | AGGACAGGTG | CCGGCAGCGC | 550 |
| 551 | TCTGGGTCAT | TTTCGGCGAG | GACCGCTTTC | GCTGGAGCGC | GACGATGATC | 600 |
| 601 | GGCCTGTCGC | TTGCGGTATT | CGGAATCTTG | CACGCCCTCG | CTCAAGCCTT | 650 |
| 651 | CGTCACTGGT | CCCGCCACCA | AACGTTTCGG | CGAGAAGCAG | GCCATTATCG | 700 |
| 701 | CCGGCATGGC | GGCCGACGCG | CTGGGCTACG | TCTTGCTGGC | GTTCGCGACG | 750 |
| 751 | CGAGGCTGGA | TGGCCTTCCC | CATTATGATT | CTTCTCGCTT | CCGGCGGCAT | 800 |
| 801 | CGGGATGCCC | GCGTTGCAGG | CCATGCTGTC | CAGGCAGGTA | GATGACGACC | 850 |
| 851 | ATCAGGGACA | GCTTCAAGGA | TCGCTCGCGG | CTCTTACCAG | CCTAACTTCG | 900 |
| 901 | ATCACTGGAC | CGCTGATCGT | CACGGCGATT | TATGCCGCCT | CGGCGAGCAC | 950 |
| 951 | ATGGAACGGG | TTGGCATGGA | TTGTAGGCGC | CGCCCTATAC | CTTGTCTGCC | 1000 |
| 1001 | TCCCCGCGTT | GCGTCGCGGT | GCATGGAGCC | GGGCCACCTC | GACCTGAATG | 1050 |
| 1051 | GAAGCCGGCG | GCACCTCGCT | AACGGATTCA | CCACTCCAAG | AATTGGAGCC | 1100 |
| 1101 | AATCAATTCT | TGCGGAGAAC | TGTGAATGCG | CAAACCAACC | CTTGGCAGAA | 1150 |
| 1151 | CATATCCATC | GCGTCCGCCA | TCTCCAGCAG | CCGCACGCGG | CGCATCTCGG | 1200 |
| 1201 | GCAGCGTTGG | GTCCTGGCCA | CGGGTGCGCA | TGATCGTGCT | CCTGTCGTTG | 1250 |
| 1251 | AGGACCCGGC | TAGGCTGGCG | GGGTTGCCTT | ACTGGTTAGC | AGAATGAATC | 1300 |
| 1301 | ACCGATACGC | GAGCGAACGT | GAAGCGACTG | CTGCTGCAAA | ACGTCTGCGA | 1350 |
| 1351 | CCTGAGCAAC | AACATGAATG | GTCTTCGGTT | TCCGTGTTTC | GTAAAGTCTG | 1400 |
| 1401 | GAAACGCGGA | AGTCAGCGCC | CTGCACCATT | ATGTTCCGGA | TCTGCATCGC | 1450 |
| 1451 | AGGATGCTGC | TGGCTACCCT | GTGGAACACC | TACATCTGTA | TTAACGAAGC | 1500 |
| 1501 | GCTGGCATTG | ACCCTGAGTG | ATTTTTCTCT | GGTCCCGCCG | CATCCATACC | 1550 |
| 1551 | GCCAGTTGTT | TACCCTCACA | ACGTTCCAGT | AACCGGGCAT | GTTCATCATC | 1600 |
| 1601 | AGTAACCCGT | ATCGTGAGCA | TCCTCTCTCG | TTTCATCGGT | ATCATTACCC | 1650 |
| 1651 | CCATGAACAG | AAATCCCCCT | TACACGGAGG | CATCAGTGAC | CAAACAGGAA | 1700 |
| 1701 | AAAACCGCCC | TTAACATGGC | CCGCTTTATC | AGAAGCCAGA | CATTAACGCT | 1750 |
| 1751 | TCTGGAGAAA | CTCAACGAGC | TGGACGCGGA | TGAACAGGCA | GACATCTGTG | 1800 |
| 1801 | AATCGCTTCA | CGACCACGCT | GATGAGCTTT | ACCGCAGCTG | CCTCGCGCGT | 1850 |
| 1851 | TTCGGTGATG | ACGGTGAAAA | CCTCTGACAC | ATGCAGCTCC | CGGAGACGGT | 1900 |
| 1901 | CACAGCTTGT | CTGTAAGCGG | ATGCCGGGAG | CAGACAAGCC | CGTCAGGGCG | 1950 |
| 1951 | CGTCAGCGGG | TGTTGGCGGG | TGTCGGGGCG | CAGCCATGAC | CCAGTCACGT | 2000 |
| 2001 | AGCGATAGCG | GAGTGTATAC | TGGCTTAACT | ATGCGGCATC | AGAGCAGATT | 2050 |
| 2051 | GTACTGAGAG | TGCACCATAT | ATGCGGTGTG | AAATACCGCA | CAGATGCGTA | 2100 |
| 2101 | AGGAGAAAAT | ACCGCATCAG | GCGCTCTTCC | GCTTCCTCGC | TCACTGACTC | 2150 |
| 2151 | GCTGCGCTCG | GTCGTTCGGC | TGCGGCGAGC | GGTATCAGCT | CACTCAAAGG | 2200 |
| 2201 | CGGTAATACG | GTTATCCACA | GAATCAGGGG | ATAACGCAGG | AAAGAACATG | 2250 |
| 2251 | TGAGCAAAAG | GCCAGCAAAA | GGCCAGGAAC | CGTAAAAAGG | CCGCGTTGCT | 2300 |
| 2301 | GGCGTTTTTC | CATAGGCTCC | GCCCCCCTGA | CGAGCATCAC | AAAAATCGAC | 2350 |
| 2351 | GCTCAAGTCA | GAGGTGGCGA | AACCCGACAG | GACTATAAAG | ATACCAGGCG | 2400 |
| 2401 | TTTCCCCCTG | GAAGCTCCCT | CGTGCGCTCT | CCTGTTCCGA | CCCTGCCGCT | 2450 |
| 2451 | TACCGGATAC | CTGTCCGCCT | TTCTCCCTTC | GGGAAGCGTG | GCGCTTTCTC | 2500 |
| 2501 | ATAGCTCACG | CTGTAGGTAT | CTCAGTTCGG | TGTAGGTCGT | TCGCTCCAAG | 2550 |
| 2551 | CTGGGCTGTG | TGCACGAACC | CCCCGTTCAG | CCCGACCGCT | GCGCCTTATC | 2600 |
| 2601 | CGGTAACTAT | CGTCTTGAGT | CCAACCCGGT | AAGACACGAC | TTATCGCCAC | 2650 |
| 2651 | TGGCAGCAGC | CACTGGTAAC | AGGATTAGCA | GAGCGAGGTA | TGTAGGCGGT | 2700 |
| 2701 | GCTACAGAGT | TCTTGAAGTG | GTGGCCTAAC | TACGGCTACA | CTAGAAGGAC | 2750 |
| 2751 | AGTATTTGGT | ATCTGCGCTC | TGCTGAAGCC | AGTTACCTTC | GGAAAAAGAG | 2800 |
| 2801 | TTGGTAGCTC | TTGATCCGGC | AAACAAACCA | CCGCTGGTAG | CGGTGGTTTT | 2850 |
| 2851 | TTTGTTTGCA | AGCAGCAGAT | TACGCGCAGA | AAAAAAGGAT | CTCAAGAAGA | 2900 |
| 2901 | TCCTTTGATC | TTTTCTACGG | GGTCTGACGC | TCAGTGGAAC | GAAAACTCAC | 2950 |
| 2951 | GTTAAGGGAT | TTTGGTCATG | AGATTATCAA | AAAGGATCTT | CACCTAGATC | 3000 |
| 3001 | CTTTTAAATT | AAAAATGAAG | TTTTAAATCA | ATCTAAAGTA | TATATGAGTA | 3050 |
| 3051 | AACTTGGTCT | GACAGTTACC | AATGCTTAAT | CAGTGAGGCA | CCTATCTCAG | 3100 |
| 3101 | CGATCTGTCT | ATTTCGTTCA | TCCATAGTTG | CCTGACTCCC | CGTCGTGTAG | 3150 |
| 3151 | ATAACTACGA | TACGGGAGGG | CTTACCATCT | GGCCCCAGTG | CTGCAATGAT | 3200 |
| 3201 | ACCGCGAGAC | CCACGCTCAC | CGGCTCCAGA | TTTATCAGCA | ATAAACCAGC | 3250 |
| 3251 | CAGCCGGAAG | GGCCGAGCGC | AGAAGTGGTC | CTGCAACTTT | ATCCGCCTCC | 3300 |
| 3301 | ATCCAGTCTA | TTAATTGTTG | CCGGGAAGCT | AGAGTAAGTA | GTTCGCCAGT | 3350 |
| 3351 | TAATAGTTTG | CGCAACGTTG | TTGCCATTGC | TGCAGGCATC | GTGGTGTCAC | 3400 |
| 3401 | GCTCGTCGTT | TGGTATGGCT | TCATTCAGCT | CCGGTTCCCA | ACGATCAAGG | 3450 |
| 3451 | CGAGTTACAT | GATCCCCCAT | GTTGTGCAAA | AAAGCGGTTA | GCTCCTTCGG | 3500 |
| 3501 | TCCTCCGATC | GTTGTCAGAA | GTAAGTTGGC | CGCAGTGTTA | TCACTCATGG | 3550 |
| 3551 | TTATGGCAGC | ACTGCATAAT | TCTCTTACTG | TCATGCCATC | CGTAAGATGC | 3600 |
| 3601 | TTTTCTGTGA | CTGGTGAGTA | CTCAACCAAG | TCATTCTGAG | AATAGTGTAT | 3650 |
| 3651 | GCGGCGACCG | AGTTGCTCTT | GCCCGGCGTC | AACACGGGAT | AATACCGCGC | 3700 |
| 3701 | CACATAGCAG | AACTTTAAAA | GTGCTCATCA | TTGGAAAACG | TTCTTCGGGG | 3750 |
| 3751 | CGAAAACTCT | CAAGGATCTT | ACCGCTGTTG | AGATCCAGTT | CGATGTAACC | 3800 |
| 3801 | CACTCGTGCA | CCCAACTGAT | CTTCAGCATC | TTTTACTTTC | ACCAGCGTTT | 3850 |
| 3851 | CTGGGTGAGC | AAAAACAGGA | AGGCAAAATG | CCGCAAAAAA | GGGAATAAGG | 3900 |
| 3901 | GCGACACGGA | AATGTTGAAT | ACTCATACTC | TTCCTTTTTC | AATATTATTG | 3950 |
| 3951 | AAGCATTTAT | CAGGGTTATT | GTCTCATGAG | CGGATACATA | TTTGAATGTA | 4000 |
| 4001 | TTTAGAAAAA | TAAACAAATA | GGGGTTCCGC | GCACATTTCC | CCGAAAAGTG | 4050 |
| 4051 | CCACCTGACG | TCTAAGAAAC | CATTATTATC | ATGACATTAA | CCTATAAAAA | 4100 |
| 4101 | TAGGCGTATC | ACGAGGCCCT | TTCGTCTTCA | AGAA | 4134 |
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文献和实验一,酵母表达系统毕赤酵母表达系统(invitrogen公司):分泌型质粒pPIC9K,宿主菌毕赤酵母GS115分泌型质粒pPICZalpha (甲醇诱导)宿主菌毕赤酵母x33分泌型质粒pHIL-S1二,原核表达质粒表达宿主菌为BL21(DE3)系列带His标签(N端):PET5-(a)氨苄抗性带His标签(N端):PET28-(a)卡那抗性带His标签(C端):PET24-(b)卡那抗性带His标签(C端):PET21-(a)氨苄抗性带His标签(N端)+TrxA(硫氧还蛋白):PET-32
一,酵母表达系统 毕赤酵母表达系统(invitrogen公司):分泌型质粒pPIC9K, 宿主菌毕赤酵母GS115 分泌型质粒pPICZalpha (甲醇诱导)宿主菌毕赤酵母x33 分泌型质粒pHIL-S1 二,原核表达质粒 表达宿主菌为BL21(DE3)系列 带His标签(N端):PET5-(a)氨苄抗性 带His标签(N端):PET28-(a)卡那抗性 带His标签(C端):PET24-(b)卡那抗性 带His标签(C端):PET21
3.1 PGEX4T-1 PGEX4T-2 PGEX4T-3 PGEX-6P-1 PGEX-KG PGEX-2T PET3b PET5b PET11a PET14
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