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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-43.1c(+)（pET43.1c）
- 库存：
大量
- 供应商：
联迈生物
- 规格：
1ug
pET-43.1c(+)（pET43.1c）
质粒名称: pET-43.1c (+)
其他名称:
质粒大小（bp）: 7230
抗性: Ampicillin (氨苄青霉素)
质粒类型: 大肠杆菌表达载体
是否病毒: Nonviral
启动子:
稳转或瞬转: Transient
组成型或诱导性: Constitutive
表达水平: High
蛋白标签: C-HSV, N-His, C-His, N-Thrombin, N-Nus, N-EK
5′ 测序引物及序列: T7: 5′-TAATACGACTCACTATAGGG-3′
3′ 测序引物及序列: ColiDOWN: 5′-TTCACTTCTGAGTTCGGCATG-3′
GenBank号:
其他属性:
质粒图谱:
| Feature Name | Start | End | |
|---|---|---|---|
| T7_Terminal_primer | 69 | 87 | |
| rrnB_T2_terminator | 168 | 141 | |
| rrnB_T1_terminator | 343 | 300 | |
| rrnB_terminator | 466 | 309 | |
| EK | 693 | 679 | |
| S15 | 789 | 745 | |
| 6xHIS | 816 | 799 | |
| T7_transl_en_RBS | 2341 | 2325 | |
| lacO | 2386 | 2359 | |
| T7_promoter | 2404 | 2386 | |
| tet (300 – 563) | 2440 | 2703 | |
| pBRrevBam_primer | 2511 | 2492 | |
| lacI | 2786 | 3877 | |
| ROP | 4449 | 4640 | |
| pGEX_3_primer | 4656 | 4634 | |
| pBR322_origin | 5674 | 5055 | |
| AmpR_promoter | 5761 | 5789 | |
| Ampicillin | 5832 | 6692 |
| ORF | Start | End | |
|---|---|---|---|
| ORF frame 1 | 427 | 2130 | |
| ORF frame 1 | 2316 | 499 | |
| ORF frame 1 | 2317 | 3045 | |
| ORF frame 2 | 2918 | 3877 | |
| ORF frame 3 | 5832 | 6692 |
| Enzyme Name | Cut |
|---|---|
| PacI | 503 |
| XhoI | 530 |
| PmlI | 576 |
| NotI | 592 |
| EagI | 592 |
| HindIII | 599 |
| KpnI | 624 |
| SalI | 626 |
| PstI | 639 |
| AscI | 641 |
| EcoRI | 654 |
| BamHI | 660 |
| SacI | 672 |
| SmaI | 719 |
| XmaI | 717 |
| SacII | 795 |
| SpeI | 826 |
| BglII | 941 |
| MscI | 1073 |
| StuI | 1126 |
| NcoI | 1401 |
| NdeI | 2315 |
| XbaI | 2353 |
| ApaI | 3356 |
| HpaI | 3651 |
| FspI | 6396 |
质粒序列:
| 1 | atccggatat | agttcctcct | ttcagcaaaa | aacccctcaa | gacccgttta | 50 |
| 51 | gaggccccaa | ggggttatgc | tagttattgc | tcagcggata | catatttgaa | 100 |
| 101 | tgtatttaga | aaaataaaca | aaagagtttg | tagaaacgca | aaaaggccat | 150 |
| 151 | ccgtcaggat | ggccttctgc | ttaatttgat | gcctggcagt | ttatggcggg | 200 |
| 201 | cgtcctgccc | gccaccctcc | gggccgttgc | ttcgcaacgt | tcaaatccgc | 250 |
| 251 | tcccggcgga | tttgtcctac | tcaggagagc | gttcaccgac | aaacaacaga | 300 |
| 301 | taaaacgaaa | ggcccagtct | ttcgactgag | cctttcgttt | tatttgatgc | 350 |
| 351 | ctggcagttc | cctactctcg | catggggaga | ccccacacta | ccatcggcgc | 400 |
| 401 | tacggcgttt | cacttctgag | ttcggcatgg | ggtcaggtgg | gaccaccgcg | 450 |
| 451 | ctactgccgc | caggcaaatt | ctgttttatc | agcctaggaa | cgcccaactt | 500 |
| 501 | aattaacatt | agtggtggtg | gtggtggtgc | tcgagatcct | ctgggtcttc | 550 |
| 551 | aggagcgagt | tctggctggc | ttgcacgtgt | atacagctgt | gcggccgcaa | 600 |
| 601 | gcttcgaacg | cgtatcgatg | gtaccgtcga | cgtcctgcag | gcgcgcctgt | 650 |
| 651 | acagaattcg | gatccacgag | ctcggactct | tgtcgtcgtc | atcaatcgta | 700 |
| 701 | ccagaacccg | cgctgccccg | ggggaccaga | ccagttggcg | gtggcgagtc | 750 |
| 751 | catgtgctgg | cgttcaaatt | tcgcagcagc | ggtttcttta | cccgcggagt | 800 |
| 801 | gatggtgatg | gtgatgacca | gaaccactag | tcgcttcgtc | accgaaccag | 850 |
| 851 | caaatattac | gggcagccat | aatcagtgct | ccggcttttt | cgtcggtcaa | 900 |
| 901 | cccttcgata | tcagccagat | catcaatgcc | ctgttcggcg | agatcttcca | 950 |
| 951 | gcgtacaaac | gccacgggcg | gccagtttga | atgccaaatc | acgatctacc | 1000 |
| 1001 | ccttcaaggt | tcagcagatc | gtcagccggt | ttgttatcac | cgaggctttc | 1050 |
| 1051 | ttcctgggcc | tgtgcaatgg | tggccagtgc | atttttagca | cgctcgcgca | 1100 |
| 1101 | gtgcttcaac | ggtcggctca | tcaaggcctt | cgatttccaa | cagctctttc | 1150 |
| 1151 | atcggcacat | aggccaattc | ttccagcgtc | gagaagcctt | cttctaccag | 1200 |
| 1201 | aacagtcgcg | aagtcttcgt | cgatgtcgag | atatttggtg | aaggtgtcga | 1250 |
| 1251 | tcgctgcgtg | cgcttccgcc | tgatgcttag | cttgcaggtc | gtcaacggtc | 1300 |
| 1301 | atcacgttga | gttcccaacc | gctcagttgc | gaagccagac | gcacgttctg | 1350 |
| 1351 | accgttacgg | ccaatcgcct | gcgccagatt | accggcttca | acggcgatgt | 1400 |
| 1401 | ccatggtgtg | tttatcttca | tccaccacga | tagaagcaac | gtctgccggt | 1450 |
| 1451 | gccattgcgt | taatcacgaa | ctgcgccggg | ttatcatccc | acaggacgat | 1500 |
| 1501 | atcgatacgc | tcgccaccca | gttcagtaga | caccgcctga | acacgcgcgc | 1550 |
| 1551 | cacgcatacc | tacgcaagca | cctaccggat | cgatacgttt | atcgttggtt | 1600 |
| 1601 | ttcaccgcga | ttttcgcacg | agaacccgga | tcgcgagccg | ctgctttaat | 1650 |
| 1651 | ttcaatcact | tcttcgccga | tttctggcac | ttcaatacgg | aacagttcga | 1700 |
| 1701 | tcagcatttc | cggcttggaa | cgagtgacga | acagttgcgc | gccacgcgct | 1750 |
| 1751 | tccgggcgaa | cggaatagag | cacgccacga | acgcggtcgc | cagggcggaa | 1800 |
| 1801 | gttttcacgc | ggcagcatat | cttcgcgcag | gatcacggct | tcagcgttgt | 1850 |
| 1851 | tgcccagatc | cagagagatg | ttgtcgcggt | ttactttttt | caccacgccg | 1900 |
| 1901 | gtgatgattt | caccttcgtg | ttcacggaac | tgatcaacca | ccatcgcacg | 1950 |
| 1951 | ttcggcttca | cgcactttct | gcacgataac | ctgttttgcc | gtctgggtag | 2000 |
| 2001 | tgatacggtc | aaaggtaaca | gactcaatct | gatcttcaac | gtaatcgccc | 2050 |
| 2051 | aggttcaggc | tttcatcttc | ataacgtgcg | gcttcaaggg | tgatttcctt | 2100 |
| 2101 | ggtcggctgg | gtgacttcat | caacaactaa | ccagcgacgg | aaagtgtcaa | 2150 |
| 2151 | aatcaccgct | tttgcgatcg | atctgtacgc | ggacgtcgat | ctcttgttca | 2200 |
| 2201 | tattttttct | ttgttgctgt | cgccagcgcg | ctttccaatg | cttcgaaaat | 2250 |
| 2251 | cttctcgcga | ggtagcgcct | tttcattgga | tacggcttca | actacagcca | 2300 |
| 2301 | aaatttcttt | gttcatatgt | atatctcctt | cttaaagtta | aacaaaatta | 2350 |
| 2351 | tttctagagg | ggaattgtta | tccgctcaca | attcccctat | agtgagtcgt | 2400 |
| 2401 | attaatttcg | cgggatcgag | atcgatctcg | atcctctacg | ccggacgcat | 2450 |
| 2451 | cgtggccggc | atcaccggcg | ccacaggtgc | ggttgctggc | gcctatatcg | 2500 |
| 2501 | ccgacatcac | cgatggggaa | gatcgggctc | gccacttcgg | gctcatgagc | 2550 |
| 2551 | gcttgtttcg | gcgtgggtat | ggtggcaggc | cccgtggccg | ggggactgtt | 2600 |
| 2601 | gggcgccatc | tccttgcatg | caccattcct | tgcggcggcg | gtgctcaacg | 2650 |
| 2651 | gcctcaacct | actactgggc | tgcttcctaa | tgcaggagtc | gcataaggga | 2700 |
| 2701 | gagcgtcgag | atcccggaca | ccatcgaatg | gcgcaaaacc | tttcgcggta | 2750 |
| 2751 | tggcatgata | gcgcccggaa | gagagtcaat | tcagggtggt | gaatgtgaaa | 2800 |
| 2801 | ccagtaacgt | tatacgatgt | cgcagagtat | gccggtgtct | cttatcagac | 2850 |
| 2851 | cgtttcccgc | gtggtgaacc | aggccagcca | cgtttctgcg | aaaacgcggg | 2900 |
| 2901 | aaaaagtgga | agcggcgatg | gcggagctga | attacattcc | caaccgcgtg | 2950 |
| 2951 | gcacaacaac | tggcgggcaa | acagtcgttg | ctgattggcg | ttgccacctc | 3000 |
| 3001 | cagtctggcc | ctgcacgcgc | cgtcgcaaat | tgtcgcggcg | attaaatctc | 3050 |
| 3051 | gcgccgatca | actgggtgcc | agcgtggtgg | tgtcgatggt | agaacgaagc | 3100 |
| 3101 | ggcgtcgaag | cctgtaaagc | ggcggtgcac | aatcttctcg | cgcaacgcgt | 3150 |
| 3151 | cagtgggctg | atcattaact | atccgctgga | tgaccaggat | gccattgctg | 3200 |
| 3201 | tggaagctgc | ctgcactaat | gttccggcgt | tatttcttga | tgtctctgac | 3250 |
| 3251 | cagacaccca | tcaacagtat | tattttctcc | catgaagacg | gtacgcgact | 3300 |
| 3301 | gggcgtggag | catctggtcg | cattgggtca | ccagcaaatc | gcgctgttag | 3350 |
| 3351 | cgggcccatt | aagttctgtc | tcggcgcgtc | tgcgtctggc | tggctggcat | 3400 |
| 3401 | aaatatctca | ctcgcaatca | aattcagccg | atagcggaac | gggaaggcga | 3450 |
| 3451 | ctggagtgcc | atgtccggtt | ttcaacaaac | catgcaaatg | ctgaatgagg | 3500 |
| 3501 | gcatcgttcc | cactgcgatg | ctggttgcca | acgatcagat | ggcgctgggc | 3550 |
| 3551 | gcaatgcgcg | ccattaccga | gtccgggctg | cgcgttggtg | cggacatctc | 3600 |
| 3601 | ggtagtggga | tacgacgata | ccgaagacag | ctcatgttat | atcccgccgt | 3650 |
| 3651 | taaccaccat | caaacaggat | tttcgcctgc | tggggcaaac | cagcgtggac | 3700 |
| 3701 | cgcttgctgc | aactctctca | gggccaggcg | gtgaagggca | atcagctgtt | 3750 |
| 3751 | gcccgtctca | ctggtgaaaa | gaaaaaccac | cctggcgccc | aatacgcaaa | 3800 |
| 3801 | ccgcctctcc | ccgcgcgttg | gccgattcat | taatgcagct | ggcacgacag | 3850 |
| 3851 | gtttcccgac | tggaaagcgg | gcagtgagcg | caacgcaatt | aatgtaagtt | 3900 |
| 3901 | agctcactca | ttaggcaccg | ggatctcgac | cgatgccctt | gagagccttc | 3950 |
| 3951 | aacccagtca | gctccttccg | gtgggcgcgg | ggcatgacta | gcatgatcgt | 4000 |
| 4001 | gctcctgtcg | ttgaggaccc | ggctaggctg | gcggggttgc | cttactggtt | 4050 |
| 4051 | agcagaatga | atcaccgata | cgcgagcgaa | cgtgaagcga | ctgctgctgc | 4100 |
| 4101 | aaaacgtctg | cgacctgagc | aacaacatga | atggtcttcg | gtttccgtgt | 4150 |
| 4151 | ttcgtaaagt | ctggaaacgc | ggaagtcagc | gccctgcacc | attatgttcc | 4200 |
| 4201 | ggatctgcat | cgcaggatgc | tgctggctac | cctgtggaac | acctacatct | 4250 |
| 4251 | gtattaacga | agcgctggca | ttgaccctga | gtgatttttc | tctggtcccg | 4300 |
| 4301 | ccgcatccat | accgccagtt | gtttaccctc | acaacgttcc | agtaaccggg | 4350 |
| 4351 | catgttcatc | atcagtaacc | cgtatcgtga | gcatcctctc | tcgtttcatc | 4400 |
| 4401 | ggtatcatta | cccccatgaa | cagaaatccc | ccttacacgg | aggcatcagt | 4450 |
| 4451 | gaccaaacag | gaaaaaaccg | cccttaacat | ggcccgcttt | atcagaagcc | 4500 |
| 4501 | agacattaac | gcttctggag | aaactcaacg | agctggacgc | ggatgaacag | 4550 |
| 4551 | gcagacatct | gtgaatcgct | tcacgaccac | gctgatgagc | tttaccgcag | 4600 |
| 4601 | ctgcctcgcg | cgtttcggtg | atgacggtga | aaacctctga | cacatgcagc | 4650 |
| 4651 | tcccggagac | ggtcacagct | tgtctgtaag | cggatgccgg | gagcagacaa | 4700 |
| 4701 | gcccgtcagg | gcgcgtcagc | gggtgttggc | gggtgtcggg | gcgcagccat | 4750 |
| 4751 | gacccagtca | cgtagcgata | gcggagtgta | tactggctta | actatgcggc | 4800 |
| 4801 | atcagagcag | attgtactga | gagtgcacca | tatatgcggt | gtgaaatacc | 4850 |
| 4851 | gcacagatgc | gtaaggagaa | aataccgcat | caggcgctct | tccgcttcct | 4900 |
| 4901 | cgctcactga | ctcgctgcgc | tcggtcgttc | ggctgcggcg | agcggtatca | 4950 |
| 4951 | gctcactcaa | aggcggtaat | acggttatcc | acagaatcag | gggataacgc | 5000 |
| 5001 | aggaaagaac | atgtgagcaa | aaggccagca | aaaggccagg | aaccgtaaaa | 5050 |
| 5051 | aggccgcgtt | gctggcgttt | ttccataggc | tccgcccccc | tgacgagcat | 5100 |
| 5101 | cacaaaaatc | gacgctcaag | tcagaggtgg | cgaaacccga | caggactata | 5150 |
| 5151 | aagataccag | gcgtttcccc | ctggaagctc | cctcgtgcgc | tctcctgttc | 5200 |
| 5201 | cgaccctgcc | gcttaccgga | tacctgtccg | cctttctccc | ttcgggaagc | 5250 |
| 5251 | gtggcgcttt | ctcatagctc | acgctgtagg | tatctcagtt | cggtgtaggt | 5300 |
| 5301 | cgttcgctcc | aagctgggct | gtgtgcacga | accccccgtt | cagcccgacc | 5350 |
| 5351 | gctgcgcctt | atccggtaac | tatcgtcttg | agtccaaccc | ggtaagacac | 5400 |
| 5401 | gacttatcgc | cactggcagc | agccactggt | aacaggatta | gcagagcgag | 5450 |
| 5451 | gtatgtaggc | ggtgctacag | agttcttgaa | gtggtggcct | aactacggct | 5500 |
| 5501 | acactagaag | gacagtattt | ggtatctgcg | ctctgctgaa | gccagttacc | 5550 |
| 5551 | ttcggaaaaa | gagttggtag | ctcttgatcc | ggcaaacaaa | ccaccgctgg | 5600 |
| 5601 | tagcggtggt | ttttttgttt | gcaagcagca | gattacgcgc | agaaaaaaag | 5650 |
| 5651 | gatctcaaga | agatcctttg | atcttttcta | cggggtctga | cgctcagtgg | 5700 |
| 5701 | aacgaaaact | cacgttaagg | gattttggtc | atgacctatt | tgtttatttt | 5750 |
| 5751 | tctaaataca | ttcaaatatg | tatccgctca | tgagacaata | accctgataa | 5800 |
| 5801 | atgcttcaat | catgattgaa | aaaggaagag | tatgagtatt | caacatttcc | 5850 |
| 5851 | gtgtcgccct | tattcccttt | tttgcggcat | tttgccttcc | tgtttttgct | 5900 |
| 5901 | cacccagaaa | cgctggtgaa | agtaaaagat | gctgaagatc | agttgggtgc | 5950 |
| 5951 | acgagtgggt | tacatcgaac | tggatctcaa | cagcggtaag | atccttgaga | 6000 |
| 6001 | gttttcgccc | cgaagaacgt | tttccaatga | tgagcacttt | taaagttctg | 6050 |
| 6051 | ctatgtggcg | cggtattatc | ccgtattgac | gccgggcaag | agcaactcgg | 6100 |
| 6101 | tcgccgcata | cactattctc | agaatgactt | ggttgagtac | tcaccagtca | 6150 |
| 6151 | cagaaaagca | tcttacggat | ggcatgacag | taagagaatt | atgcagtgct | 6200 |
| 6201 | gccataacca | tgagtgataa | cactgcggcc | aacttacttc | tgacaacgat | 6250 |
| 6251 | cggaggaccg | aaggagctaa | ccgctttttt | gcacaacatg | ggggatcatg | 6300 |
| 6301 | taactcgcct | tgatcgttgg | gaaccggagc | tgaatgaagc | cataccaaac | 6350 |
| 6351 | gacgagcgtg | acaccacgat | gcctgtagca | atggcaacaa | cgttgcgcaa | 6400 |
| 6401 | actattaact | ggcgaactac | ttactctagc | ttcccggcaa | caattaatag | 6450 |
| 6451 | actggatgga | ggcggataaa | gttgcaggac | cacttctgcg | ctcggccctt | 6500 |
| 6501 | ccggctggct | ggtttattgc | tgataaatct | ggagccggtg | agcgtgggtc | 6550 |
| 6551 | tcgcggtatc | attgcagcac | tggggccaga | tggtaagccc | tcccgtatcg | 6600 |
| 6601 | tagttatcta | cacgacgggg | agtcaggcaa | ctatggatga | acgaaataga | 6650 |
| 6651 | cagatcgctg | agataggtgc | ctcactgatt | aagcattggt | aactgtcaga | 6700 |
| 6701 | ccaagtttac | tcatatatac | tttagattga | tttaaaactt | catttttaat | 6750 |
| 6751 | ttaaaaggat | ctaggtgaag | atcctttttg | ataatctcat | gccatcgtgc | 6800 |
| 6801 | cgccagaaat | tgtaaacgtt | aatattttgt | taaaattcgc | gttaaatttt | 6850 |
| 6851 | tgttaaatca | gctcattttt | taaccaatag | gccgaaatcg | gcaaaatccc | 6900 |
| 6901 | ttataaatca | aaagaataga | ccgagatagg | gttgagtgtt | gttccagttt | 6950 |
| 6951 | ggaacaagag | tccactatta | aagaacgtgg | actccaacgt | caaagggcga | 7000 |
| 7001 | aaaaccgtct | atcagggcga | tggcccacta | cgtgaaccat | caccctaatc | 7050 |
| 7051 | aagttttttg | gggtcgaggt | gccgtaaagc | actaaatcgg | aaccctaaag | 7100 |
| 7101 | ggagcccccg | atttagagct | tgacggggaa | agccggcgaa | cgtggcgaga | 7150 |
| 7151 | aaggaaggga | agaaagcgaa | aggagcgggc | gctagggcgc | tggcaagtgt | 7200 |
| 7201 | agcggtcacg | ctgcgcgtaa | ccaccacacc | 7230 |
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文献和实验相关实验
加热灭活酶。通过凝胶提取试剂盒纯化消化的pET-32α（+）载体骨架和从pMD18-T载体切下的基因片段。用T4 DNA连接酶在23℃下用pET-32α（+）载体骨架连接基因片段1小时，载体末端和插入末端的比例为1：3（fmol）。将10μl连接反应与100μl感受态大肠杆菌 BL21（DE3）在冰上混合10分钟，并将混合物在42℃下孵育90秒。加入800μlLB并在37°C下孵育40分钟。涂在含有100μg/ ml氨苄青霉素的LB平板上。在37°C孵育过夜。通过菌落PCR筛选插入物的存在或制备
Purpose Materials10ml 6% dextran + 7ml citrate/citric acidDextran: T500 --> 6g+100ml PBSCitrate solution: 25g Na Citrate + 8g citric acid + 500 ml PBS 43 ml blood 12 ml RT Histopaque 107718 ml cold H2 O2 ml 10x PBSM199 for HUVECs: 1L powder pocket
PET39b PET40b PET41a PET42a PET42b PET43.1b PET44a PET48b PET50b PET52b
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pET-43.1c(+)（pET43.1c）
¥1 - 1880
