6 附录
6.1.1常用抗生素配方
6.1.2常用母液配方
6.1.3缓冲液(pH 6-10)的pKa及缓冲范围
6.1.4磷酸缓冲液配方
6.2.1常用网站
6.2.2通用测序引物
6.2.3密码子及其偏好性表格
6.2.4空间群表格
6.2.5线站新衍射仪BL17U1
6.2.6实验室载体
6.2.7实验室细胞株及菌株
6.2.8结晶试剂盒
抗生素 | 母液浓度(mg/mL) | 工作浓度(μg/mL)for 严紧型质粒 | 工作浓度(μg/mL)for 松驰型质粒 | 处理方式 | |
Ampicillin,氨苄青霉素(Amp) | 100(溶于水) | 20 | 60 | 过滤除菌 | |
Kanamycin,卡那霉素(Kan) | 50 (溶于水) | 10 | 50 | 过滤除菌/高压 | |
Chloramphenicol,氯霉素(Chl/Cm/CAP) | 34 (溶于乙醇)
| 25 | 170 | 无需灭菌 | |
Carbenicillin,羧苄青霉素(Cab) | 50 (溶于水) | 20 | 60 | 过滤除菌 | |
Streptomycin,链霉素(Sm) | 10 (溶于水) | 10 | 50 | 过滤除菌 | |
Tetracyyline,四环素(Tet) | 5 (溶于甲醇) | 10 | 50 | 无需灭菌 | |
1. 保存条件:建议 -20 °C保存。
2. mycin,霉素,在不严格的缩写中作为“XX霉素”的后缀缩写为m。
3. 氯霉素(Chlor Am Phenicol),氯氨苯醇,标准缩写CAP,有时也写做Cm。
4. 过滤灭菌:用0.22 μm一次性滤膜过滤除菌,于超净台内。
5. 母液配制方法举例,如100 mg/mL氨苄青霉素:溶解1 g氨苄青霉素钠盐于足量的水中,最后定容至10 mL。分装成小份于-20 °C贮存。常以25 μg/mL~50 μg/mL的终浓度添加于生长培养基
1. 50× TAE(1 L):
Tris 242 g
Glacial Acetic Acid (冰醋酸) 57.1 mL
0.5 M EDTA 100 mL
2. 10× TBE(1 L):
Tris 108.8 g
Boric Acid 55.0 g
0.5 M EDTA 40 mL
3. 6×DNA Loading Buffer(100 mL):
1 M Tris-HCl pH 7.4 10 mL
100% Glycerol 60 mL
0.5 M EDTA 12 mL
Bromophenol Blue 0.01 g
4. 0.5 M EDTA pH 8.0(500 mL):
EDTA Disodium Salt Dihydrate 93.05 g
Dissolve with about 700 mL ddH2O, stir, then adjust pH to 8.0 (1 M NaOH)
Adjust the volume to 1 L. Filtered with 0.22 μm membrane or autoclave.
**EDTA does not dissolve until nearly pH 8.0
5. X-Gal (40 mg/mL), 1000×(5 mL)
X-Gal 0.200 g
Dimethyl Foramide 5 mL
(use glass pipette!)
Make ~500 μL aliquots. Store in brown glass vials at -20 °C.
6. SOC medium(250 mL)
Tryptone Peptone 5 g
Yeast extract 1.25 g
NaCl 0.15 g
KCl(10 mM) 0.05 g
MgCl2(10 mM) 0.51 g
50% glucose 4 mL
add sterile and MgCl2 and glucose after autoclaving
7. Solutions for Plasmid Preparations
1) Qiagen Buffer P1:50 mM Tris-HCl pH 8.0,10 mM EDTA,100 μg/mL RNaseA
&:for resuspension of cells in DNA plasmid preparation. 和生工P1通用。
1 M Tris pH 8.0 5 mL
0.5 M EDTA pH 8.0 2 mL
add ddH2O to 100 mL, Autoclave. Rnase A (add after autoclaving) 10.0 mg. Store at 4 °C
2) Qiagen Buffer P2:200 mM NaOH,1% SDS
&:for lysis of cells in DNA plasmid preparation. 和生工P2通用。
NaOH 0.8 g
SDS 1.0 g
add ddH2O to 100 mL
3) Qiagen Buffer P3:3.0 M potassium acetate pH 5.5,
&:not for spin columns, but for Qiatips, midi, maxi, giga kits
Potassium Acetate 25.03 g
Acetic Acid, Galacial ~11.5 mL
Adjust pH to 5.5, add ddH2O to 100 mL
4) Qiagen Buffer N3:4.2 M Gu-HCl,0.9 M potassium acetate pH 4.8
&:as nutrealization buffer to precipitate proteins and genomic materials,和生工小抽试剂盒P3通用。
5) Qiagen Buffer PB(5 M Gu-HCl, 30% isopropanol)=PBI(I for pH indicator)
6) Qiagen Buffer QG(5.5 M guanidine thiocyanate (GuSCN),20 mM Tris HCl pH 6.6)
7) Qiagen Buffer PE(10 mM Tris-HCl pH 7.5,80% ethanol )
&:for spin column salt wash in miniprep (blue)or pink columns. 用于质粒小抽和核酸提取离心柱清洗除盐。
8) Qiagen Buffer QX1:7 M NaPO4,10 mM NaAc pH 5.3
&:for solution and binding of agarose gels,用在用琼脂糖珠子DNA纯化试剂盒。
9) Qiagen Buffer QXB:5 M GuHCl
&:for binding of large >3000 bp fragments to columns
10) Qiagen Buffer QBT:750 mM NaCl,50 mM MOPS pH 7.0,15% isopropanol,0.15% triton X-100
&:Equilibration buffer
NaCl 4.38 g
Isopropanol 15 mL
MOPS 1.05 g
Triton X-100 150 mL
Adjust pH to 7.0(use 1N), add ddH2O to 100 mL
11) Qiagen Buffer QC:1.0 M NaCl,50 mM MOPS pH 7.0,15% isopropanol
&:Wash buffer
NaCl 5.84 g
MOPS 1.05 g
Isopropanol 15 mL
Adjust pH to 7.0, add ddH2O to 100 mL ,filter sterilize.
12) Qiagen Buffer QF:1.25 M NaCl, 50 mM Tris-HCl pH 8.5,15% isopropanol
&:Elution buffer
NaCl 7.31 g
Tris 8.5 1 M 5 mL
Isopropanol 15 mL
Adjust pH to 8.5, add ddH2O to 100 mL
8. Buffers for Protein Electrophoresis
1) 6×SDS Sample Buffer
1 M Tris-HCl pH 6.8 25 mL
10% SDS 20 mL
50% Glycerol 50 mL
1 M DTT 2 mL
Bromophenol Blue 0.01 g
Add ddH2O to 100 mL
2) Coommassie Blue Staining Solution(500 mL):
Coommassie Brilliant Blue R250 0.5 g
Ethanol 200 mL
Acetic acid: 50 mL
ddH2O to 500 mL(~250 mL)
(Dissolve Coommassie in Ethanol before adding water and acetic acid will require stirring 1~2 hours)
3) Coommassie Blue Destaining Solution (500 mL):
Ethanol 200 mL
Acetic Acid 50 mL
ddH2O 250 mL
4) 20×MES-SDS running buffer (1 L):MES:2-(N-吗啉)乙磺酸一水合物
&:For high-resolution separation of small size proteins.
MES 195.2 g
Tris 121.2 g
SDS 20 g
EDTA 6 g
9. Molecular Biology Buffers:
1) A1:1×NEBuffer 1(yellow):10 mM Bis Tris Propane-HCl,10 mM MgCl2,1 mM DTT pH 7.0 @ 25 °C。Supplied as a 10×concentrated stock.
2) A2:1×NEBuffer 2(blue):50 mM NaCl, 10 mM Tris-HCl,10 mM MgCl2,1 mM DTT pH 7.9 @ 25 °C。Supplied as a 10×concentrated stock.
3) A2:1×NEBuffer 3(red):100 mM NaCl, 50 mM Tris-HCl,10 mM MgCl2,1 mM DTT pH 7.9 @ 25 °C。Supplied as a 10×concentrated stock.
4) A2:1×NEBuffer 4(green,cutsmart buffer):50 mM potassium acetate,20 mM Tris acetate,10 mM magnesium acetate,1 mM DTT, pH 7.9 @ 25 °C。Supplied as a 10×concentrated stock.
5) 2×T4 DNA quick ligase 快连接酶buffer:60 mM Tris-HCl pH 7.8,20 mM MgCl2,2 mM DTT,2 mM ATP and 10% PEG6000
6) 10×T4 DNA连接酶buffer:500 mM Tris-HCl pH 7.6,100 mM MgCl2,100 mM DTT,10 mM ATP
7) 10×pfu buffer:200 mM Tris-HCl pH 8.6,100 mM KCl,160 mM (NH4)2SO4,10 mM MgCl2,1% Triton X-100,1 mg/mL BSA。
8) 5×HF buffer:32 mM HEPES-KOH buffer pH 7.8,100 mM KAcO, 4 mM Mg (AcO) 2,0.05% BSA(0.5 mg/mL)
9) 5×GC rich buffer:32 mM HEPES-KOH buffer pH 7.8,100 mM KAcO,7.5 mM Mg(AcO)2,0.01% BSA(0.1 mg/mL),1.0% DMSO
10) 2×Taq mix:
1 M Tris-HCl pH 8.6 4 mL
1 M KCl 20 mL
1 M MgCl2 600 μL
Sucrose 68 g
dNTP (2 mM each) 40 mL
1% Cresol Red(甲酚红) 2 mL
Taq polymerase (2U/μL) 20 mL
Add ddH2O to 200 mL
11) 10×Taq buffer:
200 mM Tris-HCl 8.4,200 mM KC,100 mM (NH4)2SO4,30 mM MgSO4,1 mg/mL BSA
6.1.3 缓冲液(pH 6-10)的pKa及缓冲范围
缓冲液 | pKa(25°C) | 缓冲范围 | 分子量 | 中文全称 |
MES | 6.1 | 5.5~6.7 | 195.2 | 2-吗啉乙磺酸 |
Bis-Tris | 6.5 | 5.8~7.2 | 209.2 | 二(2-羟乙基)亚胺基三(羟甲基)甲烷 |
ADA | 6.6 | 6.0~7.2 | 190.2 | N-(氨基甲酰基甲基)亚氨基二乙酸 |
ACES | 6.8 | 6.1~7.5 | 182.2 | N-氨基甲酰甲基乙磺酸 |
PIPES | 6.8 | 6.1~7.5 | 302.4 | 哌嗪-N,N'-二(2-乙磺酸) |
MOPSO | 6.9 | 6.2~7.6 | 225.3 | 3-(N-吗啉基)-2-羟基丙磺酸 |
Bis-Tris Propane | 6.8 | 6.3~9.5 | 282.3 | 双[三(羟甲基)氨基丙烷]/1,3-二[三(羟甲基)甲氨基]丙烷 |
BES | 7.1 | 6.4~7.8 | 213.2 | N-双(2-羟乙基)-2-氨基乙璜酸 |
MOPS | 7.2 | 6.5~7.9 | 209.3 | 3-(N-吗啡啉)乙磺酸 |
HEPES | 7.5 | 6.8~8.2 | 238.3 | N-2-羟乙基哌嗪-N'-2-乙磺酸 |
TES | 7.4 | 6.8~8.2 | 229.2 | N-3-(羟甲基)甲基-2-氨基乙磺酸 |
DIPSO | 7.6 | 7.0~8.2 | 243.3 | 3-[N-N-双(2-羟乙基)氨基]-2-羟基丙磺酸 |
TAPSO | 7.6 | 7.0~8.2 | 259.3 | N-3-(羟甲基)甲氨基-2-羟基丙烷磺酸 |
Tris | 8.1 | 7.0~9.1 | 121.1 | 三羟甲基氨基甲烷 |
HEPPSO | 7.8 | 7.1~8.5 | 268.3 | 4-(2-羟乙基)哌嗪-1-2-羟基丙磺酸 |
POPSO | 7.8 | 7.2~8.5 | 362.4 | 哌嗪-1,4-二羟基丙磺酸 |
EPPS | 8.0 | 7.3~8.7 | 252.3 | 4-羟乙基哌嗪丙磺酸 |
TEA | 7.8 | 7.3~8.3 | 149.2 | 三乙醇胺 |
Tricine | 8.1 | 7.4~8.8 | 179.2 | N-三-(羟甲基)甲基氨基乙酸 |
Bicine | 8.3 | 7.6~9.0 | 163.2 | N,N-双(2-羟乙基)甘氨酸 |
TAPS | 8.4 | 7.7~9.1 | 243.3 | 三羟甲基甲胺基丙磺酸 |
AMPSO | 9.0 | 8.3~9.7 | 227.3 | 3-[N-(1,1-二甲基-2-羟乙基)]氨基-2-羟丙烷磺酸 |
CHES | 9.3 | 8.6~10.0 | 207.3 | 2-(环已胺)-1-乙磺酸 |
CAPSO | 9.6 | 8.9~10.3 | 237.3 | 3-(环已胺)-2-羟基-1-丙磺酸 |
AMP | 9.7 | 9.0~10.5 | 89.1 | 2-氨基-2-甲基-1-丙醇 |
CAPS | 10.4 | 9.7~11.1 | 221.3 | 3-(环已胺)-1-丙磺酸 |
材料(MATERIALS)
r 试剂(REAGENTS)
NaH2PO4·2H2O、Na2HPO4·12H2O
配制方法:
配制时,常先配制0.2 M的NaH2PO4和0.2 M的Na2HPO4,两者按一定比例混合即成0.2 M的磷酸盐缓冲液(PB),根据需要可配制不同浓度的PB和PBS。
1. 0.2 M的 NaH2PO4:称取NaH2PO4.2H2O 31.21 g(或 NaH2PO4·H2O 27.6 g)加重蒸水至1000 mL溶解。
2. 0.2 M的 Na2HPO4:称取Na2HPO4·12H2O 71.64 g(或 Na2HPO4·7H2O 53.6 g或 Na2HPO4·2H2O 35.61 g)加重蒸水至1000 mL溶解。
3. 0.2 M pH 7.4的PB的配制:取19 mL 0.2 M的NaH2PO4和81 mL 0.2 M Na2HPO4,充分混合即为0.2 M的PB(pH约为7.4~7.5)。若pH偏高或偏低,可通过改变二者的比例来加以调整,室温保存即可。
► 25 °C下0.2 M磷酸钠缓冲液的配制
pH值 | 0.2 M Na2HPO4(mL) | 0.2 M NaH2PO4(mL) |
5.8 | 8.0 | 92.0 |
5.9 | 10.0 | 90.0 |
6.0 | 12.3 | 87.7 |
6.1 | 15.0 | 85.0 |
6.2 | 18.5 | 81.5 |
6.3 | 22.5 | 77.5 |
6.4 | 26.5 | 73.5 |
6.5 | 31.5 | 68.5 |
6.6 | 37.5 | 62.5 |
6.7 | 43.5 | 56.5 |
6.8 | 49.0 | 51.0 |
6.9 | 55.0 | 45.0 |
7.0 | 61.0 | 39.0 |
7.1 | 67.0 | 33.0 |
7.2 | 72.0 | 28.0 |
7.3 | 77.0 | 23.0 |
7.4 | 81.0 | 19.0 |
7.5 | 84.0 | 16.0 |
7.6 | 87.0 | 13.0 |
7.7 | 89.5 | 10.5 |
7.8 | 91.5 | 8.5 |
7.9 | 93.5 | 6.50 |
8.0 | 94.7 | 5.3 |
► 25 °C下0.1 M磷酸钾缓冲液的配制
pH值 | 1 M K2HPO4(mL) | 1 M KH2PO4(mL) |
5.8 | 8.5 | 91.5 |
6.0 | 13.2 | 86.8 |
6.2 | 19.2 | 80.8 |
6.4 | 27.8 | 72.2 |
6.6 | 38.1 | 61.9 |
6.8 | 49.7 | 50.3 |
7.0 | 61.5 | 38.5 |
7.2 | 71.7 | 28.3 |
7.4 | 80.2 | 19.8 |
7.6 | 86.6 | 13.4 |
7.8 | 90.8 | 9.2 |
8.0 | 94.0 | 6.0 |
1. 分子克隆(Molecular Cloning)
1) 引物Tm值计算:http://biotools.nubic.northwestern.edu/OligoCalc.html
2) 密码子优化:https://sg.idtdna.com/CodonOpt
3) 质粒图谱分析:http://wishart.biology.ualberta.ca/PlasMapper/
4) 启动子信息:https://blog.addgene.org/plasmids-101-the-promoter-region
5) addgene(一个质粒数据库):http://www.addgene.org/vector-database/
6) NEB 内切酶buffer推荐:https://nebcloner.neb.com/#!/redigest
7) 核酸数据分析:http://biotools.nubic.northwestern.edu/OligoCalc.html
2. 蛋白表达与纯化(Protein Expression and Purification)
1) 蛋白表达菌株信息:http://wolfson.huji.ac.il/expression/bac-strains-prot-exp.html
2) 蛋白纯化:http://wolfson.huji.ac.il/purification/Purification_Protocols.html
3. 蛋白序列分析(Protein Sequence Analysis)
1) 信号肽预测:http://www.cbs.dtu.dk/services/SignalP/
2) 跨膜区预:TMpred http://www.ch.embnet.org/software/TMPRED_form.html
3) 结构域预测:http://smart.embl-heidelberg.de/
4) 蛋白性质预测:http://web.expasy.org/protparam/
http://biotools.nubic.northwestern.edu/proteincalc.html
5) 蛋白质结构与功能预测:http://raptorx.uchicago.edu/
6) 二级结构预测:http://bioinf.cs.ucl.ac.uk/psipred/
7) 评估结构预测的准确性和可靠性:https://www.cameo3d.org/
8) 蛋白质结构和功能预测:https://open.predictprotein.org/
9) 三维结构比对http://ekhidna.biocenter.helsinki.fi/dali_server/start
10) 同源建模https://zhanglab.ccmb.med.umich.edu/I-TASSER/
11) 同源建模:https://swissmodel.expasy.org/
12) 预测蛋白相互作用:https://string-db.org/
13) Pfam数据库是蛋白质家族的集合(根据结构域分类):http://pfam.xfam.org/
4. 结构分析(Structure Analysis)
1) RCSB PDB 数据库:http://www.rcsb.org/
2) 晶体界面分析 PISA serverhttp://www.ebi.ac.uk/pdbe/pisa/
3) Deposition serverhttps://deposit-1.wwpdb.org/
4) 配体结构文件下载:http://www.rcsb.org/pdb/ligand/chemAdvSearch.do
5) Bernhard Rupp的生物大分子晶体学网站:http://www.ruppweb.org/Xray/101index.html
5. 其他
1) ATCC:http://www.attc.org/
2) 化学分子性质:http://www.chemicalbook.com/ProductIndex_EN.aspx
3) RNA修饰数据库:http://mods.rna.albany.edu/mods/modifications/
4) Wikipedia:https://www.wikipedia.org/
5) 各类protocols:http://openwetware.org/wiki/Protocols
6) 文献查询:https://sci-hub.tw/
6.2.2 通用测序引物
简介(INTRODUCTION)
当进行菌落PCR时,如果目的片段没有成功转化至细菌中,针对插入片段的特异性引物可能会与细菌自身DNA进行非特异性结合,产生非特异性条带,从而对我们鉴定克隆是否成功构建产生干扰。因此我们常用载体上的引物进行鉴定。利用这种引物,我们既可以检测转入的质粒是否是空载体,也能估测连入载体的插入片段大小是否正确。
以下是我们实验室常用的几个通用引物,既可以做菌落PCR,也可以用于测序。
Primer name | Site | Sequence |
Z5 | T7 promoter | TACGACTCACTATAGGGGAATTGTGAGCG |
Z6 | T7 terminator | |
Z8 | TEV and Sal I | gagaatctttatttccaaggttctgTCGAC |
Z9 | v52 Rev | GGCCAGTGAATTGTAATACGACTCACTATA |
Z10 | v55 Rev | GTTCTGATTTAATCTGTATCAGGCTGAAAA |
Z11 | pCold (v113, v114) Rev | aaatggcagggatcttagattctgtg |
XY27 | pcDNA3.1 For | taatacgactcactatagggagacccaagc |
XY28 | pcDNA3.1 Rev | tagaaggcacagtcgaggctgatca |
y936 | v108 Rev | ggctgattatgatcctctagtacttc |
Z12 | v104 For | aatttgtaatacgactcactatagggcga |
Z13 | v104 Rev | tttagaggccccaaggggttatgctagtt |
Z14 | v105 For | agatctttaatacgactcactatagggcga |
Z15 | v105 Rev | ttgagatggtgcacgatgcacagttgaa |
Z16 | V126, 127 For | agaacccactgcttactggcttatcgaaat |
Z17 | V126, 127 Rev | ctgatcagcgggtttaaactcaatggtgat |
L22 | pEGFP-N1 For | tctatataagcagagctggtttagtgaaccgtcag |
L23 | pEGFP-N1 Rev | tgcacgccgtaggtcagggtggtcac |
L28 | pCDH For | GAGCTCGTTTAGTGAACCGTCAGATCGCCT |
L29 | pCDH Rev | GTTCAATTGCCGACCCCTCCCCCCAACTTC |
第一位碱基 | 第二位碱基 | |||
U | C | A | G | |
U | UUU (Phe/F)苯丙氨酸 | UCU (Ser/S)丝氨酸 | UAU (Tyr/Y)酪氨酸 | UGU (Cys/C)半胱氨酸 |
UUC (Phe/F)苯丙氨酸 | UCC (Ser/S)丝氨酸 | UAC (Tyr/Y)酪氨酸 | UGC (Cys/C)半胱氨酸 | |
UUA (Leu/L)亮氨酸 | UCA (Ser/S)丝氨酸 | UAA (终止) | UGA (终止) | |
UUG (Leu/L)亮氨酸 | UCG (Ser/S)丝氨酸 | UAG (终止) | UGG (Trp/W)色氨酸 | |
C | CUU (Leu/L)亮氨酸 | CCU (Pro/P)脯氨酸 | CAU (His/H)组氨酸 | CGU (Arg/R)精氨酸 |
CUC (Leu/L)亮氨酸 | CCC (Pro/P)脯氨酸 | CAC (His/H)组氨酸 | CGC (Arg/R)精氨酸 | |
CUA (Leu/L)亮氨酸 | CCA (Pro/P)脯氨酸 | CAA (Gln/Q)谷氨酰胺 | CGA (Arg/R)精氨酸 | |
CUG (Leu/L)亮氨酸 | CCG (Pro/P)脯氨酸 | CAG (Gln/Q)谷氨酰胺 | CGG (Arg/R)精氨酸 | |
A | AUU (Ile/I)异亮氨酸 | ACU (Thr/T)苏氨酸 | AAU (Asn/N)天冬酰胺 | AGU (Ser/S)丝氨酸 |
AUC (Ile/I)异亮氨酸 | ACC (Thr/T)苏氨酸 | AAC (Asn/N)天冬酰胺 | AGC (Ser/S)丝氨酸 | |
AUA (Ile/I)异亮氨酸 | ACA (Thr/T)苏氨酸 | AAA (Lys/K)赖氨酸 | AGA (Arg/R)精氨酸 | |
AUG (Met/M)甲硫氨酸 | ACG (Thr/T)苏氨酸 | AAG (Lys/K)赖氨酸 | AGG (Arg/R)精氨酸 | |
G | GUU (Val/V)缬氨酸 | GCU (Ala/A)丙氨酸 | GAU (Asp/D)天冬氨酸 | GGU (Gly/G)甘氨酸 |
GUC (Val/V)缬氨酸 | GCC (Ala/A)丙氨酸 | GAC (Asp/D)天冬氨酸 | GGC (Gly/G)甘氨酸 | |
GUA (Val/V)缬氨酸 | GCA (Ala/A)丙氨酸 | GAA (Glu/E)谷氨酸 | GGA (Gly/G)甘氨酸 | |
GUG (Val/V)缬氨酸 | GCG (Ala/A)丙氨酸 | GAG (Glu/E)谷氨酸 | GGG (Gly/G)甘氨酸 |
密码子偏好性表格:
人类密码子偏好性表格
第一位碱基 | 第二位碱基 | 第三位碱基 | |||
U | C | A | G | ||
U | UUU(17.6%) | UCU(15.2%) | UAU(12.2%) | UGU(10.6%) | U |
UUC(20.3%) | UCC (17.7%) | UAC(15.3%) | UGC (12.6%) | C | |
UUA(7.7%) | UCA(12.2%) | UAA(1%) | UGA (1.6%) | A | |
UUG(12.9%) | UCG(4.4%) | UAG (0.8%) | UGG (13.2%) | G | |
C
| CUU (13.2%) | CCU(17.5%) | CAU(10.9%) | CGU(4.5%) | U |
CUC (19.6%) | CCC(19.8%) | CAC(15.1%) | CGC(10.4%) | C | |
CUA (7.2%) | CCA(16.9%) | CAA(12.3%) | CGA(6.2%) | A | |
CUG(39.6%) | CCG(6.9%) | CAG(34.2%) | CGG(11.4%) | G | |
A | AUU(16%) | ACU (13.1%) | AAU(17%) | AGU(12.1%) | U |
AUC (20.8%) | ACC(18.9%) | AAC(19.1%) | AGC(19.5%) | C | |
AUA(7.5%) | ACA(15.1%) | AAA(24.4%) | AGA(12.2%) | A | |
AUG(22%) | ACG(6.1%) | AAG(31.9%) | AGG(12%) | G | |
G | GUU(11%) | GCU(18.4%) | GAU(21.8%) | GGU(10.8%) | U |
GUC(14.5%) | GCC(27.7%) | GAC(25.1%) | GGC(22.2%) | C | |
GUA (7.1%) | GCA(15.8%) | GAA(29%) | GGA(16.5%) | A | |
GUG (28.1%) | GCG(7.4%) | GAG(39.6%) | GGG(16.5%) | G |
大肠杆菌密码子偏好性表格
第一位碱基 | 第二位碱基 | 第三位碱基 | |||
U | C | A | G | ||
U | UUU(24.4%) | UCU(13.1%) | UAU(21.6%) | UGU(5.9%) | U |
UUC(13.9%) | UCC (9.7%) | UAC(11.7%) | UGC (5.5%) | C | |
UUA(17.4%) | UCA(13.1%) | UAA(2%) | UGA (1.1%) | A | |
UUG(12.9%) | UCG(8.2%) | UAG (0.3%) | UGG (13.4%) | G | |
C
| CUU (14.5%) | CCU(9.5%) | CAU(12.4%) | CGU(15.9%) | U |
CUC(9.5%) | CCC(6.2%) | CAC(7.3%) | CGC(14%) | C | |
CUA (5.6%) | CCA(9.1%) | CAA(14.4%) | CGA(4.8%) | A | |
CUG(37.4%) | CCG(14.5%) | CAG(26.7%) | CGG(7.9%) | G | |
A | AUU(29.6%) | ACU (13.1%) | AAU(29.3%) | AGU(13.2%) | U |
AUC (19.4%) | ACC(18.9%) | AAC(20.3%) | AGC(14.3%) | C | |
AUA(13.3%) | ACA(15.1%) | AAA(37.2%) | AGA(7.1%) | A | |
AUG(23.7%) | ACG(13.6%) | AAG(15.3%) | AGG(4%) | G | |
G | GUU(21.6%) | GCU(18.9%) | GAU(33.7%) | GGU(23.7%) | U |
GUC(13.1%) | GCC(21.6%) | GAC(17.9%) | GGC(20.6%) | C | |
GUA (13.1%) | GCA(23%) | GAA(35.1%) | GGA(13.6%) | A | |
GUG (19.9%) | GCG(21.1%) | GAG(19.4%) | GGG(12.3%) | G |
6.2.4 空间群表格
6.2.5 线站新衍射仪BL17U1
简介(INTRODUCTION)
上海光源BL17U1线站新衍射仪(自主研制,水平方向SOC~±0.342 μm,垂直方向SOC~±0.319 μm)和新探测器(瑞士Dectris公司的Eiger X 16 M)已经安装调试完成,11月10号开始正式对用户开放使用。但是在开放初期还需要各位老师提醒实验人员注意以下事项:
1. 由于衍射仪测角头电机比较小,如果手动上样的话很容易把位置碰歪,所以目前只能机械手上样,建议各位老师将晶体样品保存在puck里,如果没有puck,可以提前到线站借用我们线站的puck将晶体样品转移到puck里再通过机械手上样。
2. 如果在实验的过程中发生故障,一定要第一时间给值班人员打电话,不能自行去处理。
3. 建议在EIGER 16 M探测器上收数据的策略是:0.1~0.3度一张,曝光时间0.05秒一张。这样收一套360度的完整数据(3600张衍射图)大概只需要3分钟。
4. 一套3600张衍射图的数据大小是25 GB左右,建议带一个存储空间大的硬盘拷贝数据,可以边实验边拷贝数据。减少拷贝数据等待时间。
5. 数据处理建议使用XDS。
6. 目前线站的新系统还需要进一步优化调试,能量暂时固定在12.660 Kev。
Name | Backnone | Description | Cloning Frame | Fusion Tag | Sequencing Primer 1 | Sequencing Primer2 | Resistance |
|
|
V1 | pET30a | NdeI-SalI,NotI-XhoI-His6-stop | SalI-NotI | none | T7 Promoter | T7 Terminator | Kan |
|
|
V2 | pET30a | NdeI-His8-SalI,NotI-stop | SalI-NotI | none | T7 Promoter | T7 Terminator | Kan |
|
|
V3 | pET30a | NdeI-His8-GB1-TEV-SalI,NotI-stop | SalI-NotI | His8-GB1-TEV | T7 Promoter | T7 Terminator | Kan |
|
|
V4 | pET30a | NdeI-His8-GB1-TEV-SalI,NotI-XhoI-His6-stop | SalI-NotI | His8-GB1-TEV | T7 Promoter | T7 Terminator | Kan |
|
|
V5 | pET30a | NdeI-His8-TEV-SalI,NotI-stop | SalI-NotI | MGSS-His8-TEV | T7 Promoter | T7 Terminator | Kan |
|
|
V6 | pET30a | NdeI-His8-TEV-SalI,NotI-XhoI-His6-stop | SalI-NotI | MGSS-His8-TEV | T7 Promoter | T7 Terminator | Kan |
|
|
V7 | pET30a | NdeI-His8-SUMO-TEV-SalI,NotI-XhoI-stop | SalI-NotI | MGSS-His8-SUMO-TEV | T7 Promoter | T7 Terminator | Kan |
|
|
V8 | pET30a | NdeI-His8-VHb-TEV-SalI,NotI-stop | SalI-NotI | His8-VHb-TEV | T7 Promoter | T7 Terminator | Kan |
|
|
V9 | pET30a | NdeI-His8-VHb-TEV-SalI,NotI-XhoI-His6-stop | SalI-NotI | His8-VHb-TEV | T7 Promoter | T7 Terminator | Kan |
|
|
V10 | pET30a | NdeI-His8-GST-TEV-SalI,NotI-stop | SalI-NotI | His8-GST-TEV | T7 Promoter | T7 Terminator | Kan |
|
|
V11 | pET30a | NdeI-His8-GST-TEV-SalI,NotI-XhoI-His6-stop | SalI-NotI | His8-GST-TEV | T7 Promoter | T7 Terminator | Kan |
|
|
V12 | pET30a | NdeI-His7-TRX-TEV-SalI,NotI-stop | SalI-NotI | His7-TRX-TEV | T7 Promoter | T7 Terminator | Kan |
|
|
V13 | pET30a | NdeI-His7-TRX-TEV-SalI,NotI-XhoI-His6-stop | SalI-NotI | His7-TRX-TEV | T7 Promoter | T7 Terminator | Kan |
|
|
V14 | pET30a | NdeI-His8-MBP-SalI,NotI-stop | SalI-NotI | His8-MBP | T7 Promoter | T7 Terminator | Kan |
|
|
V15 | pET30a | NdeI-His8-tag2-TEV-SalI,NotI-stop | SalI-NotI | His8-tag2-TEV | T7 Promoter | T7 Terminator | Kan |
|
|
V16 | pET30a | NdeI-His8-tag2-TEV-SalI,NotI-XhoI-His6-stop | SalI-NotI | His8-tag2-TEV | T7 Promoter | T7 Terminator | Kan |
|
|
V17 | pET30a | NdeI-DsbA-TEV-SalI,NotI-XhoI-His6-stop | SalI-NotI | DsbA-TEV | T7 Promoter | T7 Terminator | Kan |
|
|
V18 | pET30a | NdeI-MBP-SalI,NotI-XhoI-His6-stop | SalI-NotI | MBP | T7 Promoter | T7 Terminator | Kan |
|
|
V19 | pET30a | NdeI-His8-NusA-TEV-SalI,NotI-stop | SalI-NotI | His8-NusA-TEV | T7 Promoter | T7 Terminator | Kan |
|
|
V20 | pET30a | NdeI-His8-GB1-TEV-SalI,NotI-Acidic Peptide-LacZ-XhoI-His6-stop | SalI-NotI | His8-GB1-TEV | T7 Promoter | T7 Terminator | Kan |
|
|
V21 | pET30a | NdeI-His8-MBP-helical linker-SalI,NotI-stop | SalI-NotI | His8-MBP-helical linker | T7 Promoter | T7 Terminator | Kan |
|
|
V22 | pMAL | NdeI-MBP-SalI,NotI-XhoI-LacZ-stop | SalI-NotI | MBP | M13R-pUC | M13F-pUC | Amp |
|
|
V23 | pMAL | NdeI-MBP-SalI,NotI-XhoI-LacZ-stop | SalI-NotI | MBP | M13R-pUC | M13F-pUC | Amp |
|
|
V24 | pMAL | NdeI-MBP-SalI,NotI-XhoI-LacZ-stop | SalI-NotI | MBP | M13R-pUC | M13F-pUC | Amp |
|
|
V25 | pMAL | NdeI-MBP-SalI,NotI-XhoI-LacZ-stop | SalI-NotI | MBP | M13R-pUC | M13F-pUC | Amp |
|
|
V26 | pMAL | NdeI-MBP-SalI,NotI-XhoI-LacZ-stop | SalI-NotI | MBP | M13R-pUC | M13F-pUC | Amp |
|
|
V27 | pMAL | NdeI-MBP-SalI,NotI-XhoI-LacZ-stop | SalI-NotI | MBP | M13R-pUC | M13F-pUC | Amp |
|
|
V28 | pET30a | NdeI-MBP-SalI,NotI-XhoI-His6-stop | SalI-NotI | MBP | T7 Promoter | T7 Terminator | Kan |
|
|
V29 | pET30a | NdeI-MBP-TEV-SalI,NotI-stop | SalI-NotI | MBP-TEV | T7 Promoter | T7 Terminator | Kan |
|
|
V29H | pET30a | NdeI-His8-MBP-TEV-SalI,NotI-stop | SalI-NotI | His8-MBP-TEV | T7 Promoter | T7 Terminator | Kan |
|
|
V30 | pET30a | NdeI-MBP-TEV-SalI,NotI-XhoI-His6-stop | SalI-NotI | MBP-TEV | T7 Promoter | T7 Terminator | Kan |
|
|
V31 | pLEXSY-Ble | NcoI-SP-SalI,NotI-His6-stop | SalI-NotI | Signal Peptide | P14 | A26 | Amp/Ble/Tet |
|
|
V32 | pLEXSY-Ble | NcoI-SP-Strep Tag-His8-TEV-SalI,NotI-His6-stop | SalI-NotI | SP-Strep Tag-His8-TEV | P14 | A26 | Amp/Ble/Tet |
|
|
V33 | pLEXSY-Ble | NcoI-SalI,NotI-His8-stop | SalI-NotI | none | P14 | A26 | Amp/Ble/Tet |
|
|
V34 | pLEXSY-Ble | NcoI-Strep Tag-SalI,NotI-His8-stop | SalI-NotI | Strep Tag | P14 | A26 | Amp/Ble/Tet |
|
|
V35 | pLEXSY-Ble | NcoI-His8-GST-TEV-SalI,NotI-stop | SalI-NotI | His8-GST-TEV | P14 | A26 | Amp/Ble/Tet |
|
|
V36 | pLEXSY-Ble | NcoI-SP-Strep-His8-TEV-SalI,NotI-His6-stop | SalI-NotI | SP-Strep-His8-TEV | P14 | A26 | Amp/Ble/Tet |
|
|
V37 | pLEXSY-Ble | NcoI-Strep-His8-TEV-SalI,NotI-His6-stop | SalI-NotI | Strep-His8-TEV | P14 | A26 | Amp/Ble/Tet |
|
|
V40 | pTT5 | NcoI-His8-GB1-TEV-SalI,NotI-stop | SalI-NotI | His8-GB1-TEV | T7 Promoter | T7 Terminator | Kan |
|
|
V41 | pTT5 | NcoI-His6-Strep-His8-TEV-SalI,NotI-stop | SalI-NotI | His6-Strep-His8-TEV | P14 | A26 | Amp/Ble/Tet |
|
|
V42 | pTT5 | NcoI-Strep-His8-TEV-SalI,NotI-stop | SalI-NotI | Strep-His8-TEV | P14 | A26 | Amp/Ble/Tet |
|
|
V51 | pFab | EcoRV-SP-Strep-His8-TEV-SalI,NotI-His6-stop | SalI-NotI | SP-Strep-His8-TEV | unknown | unknown | Amp |
|
|
V52 | pFab | EcoRV-SP-His6-TEV-SalI,NotI-stop | SalI-NotI | SP-His6-TEV | unknown | unknown | Amp |
|
|
V53 | pFab | EcoRV-SP-His6-TEV-SalI,NotI-stop | SalI-NotI | SP-His6-TEV | unknown | unknown | Amp |
|
|
V54 | pFab | EcoRV-SP-His6-TEV-SalI,NotI-Strep-stop | SalI-NotI | SP-His6-TEV | unknown | unknown | Amp |
|
|
V55 | pMAL | NdeI-MBP-TEV-SalI,NotI-stop | SalI-NotI | MBP-TEV | M13R-pUC | M13F-pUC | Amp |
|
|
V56 | pMW103 | LexA-SalI,NotI-stop | SalI-NotI | LexA | unknown | unknown | Kan | For Y2H |
|
V57 | pET30a | NdeI-hDim2-SalI,NotI-XhoI-His6-stop | SalI-NotI | hDim2 | T7 Promoter | T7 Terminator | Kan |
|
|
V59 | pET30a | NdeI-His8-TEV-SalI,NotI-Lysozyme-stop | SalI-NotI | His8-TEV | T7 Promoter | T7 Terminator | Kan |
|
|
V60 | pET30a | NdeI-Lysozyme-SalI,NotI-XhoI-His6-stop | SalI-NotI | Lysozyme | T7 Promoter | T7 Terminator | Kan |
|
|
V61 | pEU-01 | His6-TEV-SalI,NotI-stop | SalI-NotI | His6-TEV | unknown | unknown | Amp |
|
|
V62 | pET30a | NdeI-MBP-SalI,NotI-lysozyme-XhoI-His6-stop | SalI-NotI | MBP | T7 Promoter | T7 Terminator | Kan |
|
|
V63 | pET30a | NdeI-MBP-TEV-SalI,NotI-Lysozyme-XhoI-His6-stop | SalI-NotI | MBP-TEV | T7 Promoter | T7 Terminator | Kan |
|
|
V64 | pET30a | NdeI-His8-MBP-TEV-SalI,NotI-Lysozyme-stop | SalI-NotI | His8-MBP-TEV | T7 Promoter | T7 Terminator | Kan |
|
|
V65 | pJG4-5 | Gal1-B42-HA epitope-EcoRI-SalI,NotI-stop | SalI-NotI | Gal1-B42-HA epitope | unknown | unknown | Amp | For Y2H |
|
V66 | pET30a | NdeI-T4LS5-SalI,NotI-XhoI-His6-stop | SalI-NotI | T4LS5 | T7 Promoter | T7 Terminator | Kan |
|
|
V67 | pEU-01 | NdeI-His8-NcoI-TEV-SalI,NotI-stop | SalI-NotI | His8-TEV | T7 Promoter | T7 Terminator | Kan |
|
|
V71 | pET30a | NdeI-SalI,NotI-XhoI-His6-stop | SalI-NotI | none | T7 Promoter | T7 Terminator | Kan |
|
|
V72 | pET30a | NdeI-SalI,NotI-Strep-stop | SalI-NotI | none | T7 Promoter | T7 Terminator | Kan |
|
|
V77 | pBAD | NdeI-SalI,NotI-XhoI-His6-stop | SalI-NotI | none | pBAD for | pBAD rev | Amp |
|
|
V78 | pBAD-V28E | NdeI-MBP-SalI,NotI-XhoI-His6-stop | SalI-NotI | MBP | pBAD for | pBAD rev | Amp |
|
|
V79 | pBAD-V29H | NdeI-His8-MBP-TEV-SalI,NotI-stop | SalI-NotI | His8-MBP-TEV | pBAD for | pBAD rev | Amp |
|
|
V80 | pCMV-HA | HAtag-EcoRI-SalI,NotI-stop | SalI-NotI | HAtag | pCMV | M13R-20 | Amp |
|
|
V81 | pBAD-V30 | NdeI-MBP-TEV-SalI,NotI-XhoI-His6-stop | SalI-NotI | MBP-TEV | pBAD for | pBAD rev | Amp |
|
|
V82 | pBAD-V29HS | NdeI-His8-MBP-TEV-SalI,NotI-strep-stop | SalI-NotI | His-MBP-TEV | pBAD for | pBAD rev | Amp |
|
|
V83 | pASK-17plus | NdeI-Strep-TEV-EcoRI-BamHI-XhoI-SalI-NcoI-EcoRV-HindIII | MCS | Strep-TEV | pASk for | pASK rev | Amp |
|
|
V84 | pASK-V3 | NdeI-His8-GB1-TEV-SalI,NotI-stop | SalI-NotI | His8-GB1-TEV | pASk for | pASK rev | Amp |
|
|
V85 | pASK-V28E | NdeI-MBP-SalI,NotI-XhoI-His6-stop | SalI-NotI | MBP | pASk for | pASK rev | Amp |
|
|
V86 | pASK-V29H | NdeI-His8-MBP-TEV-SalI,NotI-stop | SalI-NotI | His-MBP-TEV | pASk for | pASK rev | Amp |
|
|
V87 | pASK-V30 | NdeI-MBP-TEV-SalI,NotI-XhoI-His6-stop | SalI-NotI | MBP-TEV | pASk for | pASK rev | Amp |
|
|
V88 | pASK-V29HS | NdeI-His8-MBP-TEV-SalI,NotI-Strep-stop | SalI-NotI | His-MBP-TEV | pASk for | pASK rev | Amp |
|
|
V89 | pET30a | NdeI-InfB21-eGFP-SalI,NotI-stop | SalI-NotI | InfB21-eGFP | T7 Promoter | T7 Terminator | Kan |
|
|
V90 | pET30a | NdeI-InfB21-eGFP-TEV-SalI,NotI-stop | SalI-NotI | InfB21-eGFP | T7 Promoter | T7 Terminator | Kan |
|
|
V91 | pET30a | NdeI-His8-eGFP-TEV-SalI,NotI-stop | SalI-NotI | His8-eGFP-TEV | T7 Promoter | T7 Terminator | Kan |
|
|
V92 | pET30a | NdeI-SUMO-SalI,NotI-Strep-stop | SalI-NotI | SUMO | T7 Promoter | T7 Terminator | Kan |
|
|
V93 | pET30a | NdeI-His8-MBP-TEV-GFP-SalI,NotI-Strep-stop | SalI-NotI | His8-MBP-TEV-GFP | T7 Promoter | T7 Terminator | Kan |
|
|
V94 | pET30a | NdeI-VHb-SalI,NotI-Strep-stop | SalI-NotI | VHb | T7 Promoter | T7 Terminator | Kan |
|
|
V95 | pET30a | NdeI-Tag2-SalI,NotI-Strep-stop | SalI-NotI | Tag2 | T7 Promoter | T7 Terminator | Kan |
|
|
V96 | pET30a | NdeI-MBP-SalI,NotI-XhoI-His6-stop | SalI-NotI | MBP | T7 Promoter | T7 Terminator | Kan |
|
|
V97 | pET30a | NdeI-MBP-SalI,NotI-Strep-stop | SalI-NotI | MBP | T7 Promoter | T7 Terminator | Kan |
|
|
V98 | pFab | EcoRV-SP-NdeI-His8-SalI,NotI-Strep-stop | SalI-NotI | SP-His8 | unknown | unknown | Amp |
|
|
V99 | pET30a | NdeI-MBP-SalI,NotI-XhoI-His6-stop | SalI-NotI | MBP | T7 Promoter | T7 Terminator | Kan |
|
|
V100 | pET30a | NdeI-MBP-SalI,NotI-XhoI-His6-stop | SalI-NotI | MBP | T7 Promoter | T7 Terminator | Kan |
|
|
V101 | pET30a | NdeI-GlyA-SalI,NotI-XhoI-His6-stop | SalI-NotI | GlyA | T7 Promoter | T7 Terminator | Kan |
|
|
V102 | pET30a | NdeI-GacH-SalI,NotI-XhoI-His6-stop | SalI-NotI | GacH | T7 Promoter | T7 Terminator | Kan |
|
|
V103 | pET30a | NdeI-XBP1-SalI,NotI-XhoI-His6-stop | SalI-NotI | XBP1 | T7 Promoter | T7 Terminator | Kan |
|
|
V104 | pGBKT7SN | DNA Binding-XhoI-NdeI-SalI,NotI-stop | SalI-NotI | DNA Binding | T7 Promoter | ADH | Kan |
|
|
V105 | pGADT7SN | SV40-AD-NcoI-Epitope-NdeI-SalI,NotI-stop | SalI-NotI | SV40-AD-Epitope | T7 Promoter | ADH | Amp |
|
|
V106 | pRmHa3 | EcoRI-TEV-SalI,NotI-XhoI-His6-stop | SalI-NotI | TEV | pRmHa3 for | pRmHa3 rev | Amp |
|
|
V107 | pVL1392 | NcoI-NotI-HIs6-stop | unknown | none | y326 polyhedrin | y327 polyhedrin | Amp |
|
|
V108 | plSUMOstar | HIs6-SUMOstar-SalI,NotI-stop | SalI-NotI | SUMOstar | unknown | unknown | Amp/Get |
|
|
V109 | pFastBacHT | NdeI-InfB21-His8-TEV-SalI,NotI-stop | SalI-NotI | InfB21-His8-TEV | pASk for | pASK rev | Amp/Get |
|
|
V110 | pFastBacHT | NdeI-SalI,NotI-XhoI-His6-stop | SalI-NotI | none | pASk for | pASK rev | Amp/Get |
|
|
V111 | pFastBacHT | NdeI-His8-MBP-TEV-Sali,NotI-stop | SalI-NotI | His8-MBP-TEV | pASk for | pASK rev | Amp/Get |
|
|
V112 | pFastBacHT | NdeI-His8-MBP-TEV-Sali,NotI-XhoI-His6-stop | SalI-NotI | His8-MBP-TEV | pASk for | pASK rev | Amp/Get |
|
|
V113 | pColdII | NdeI-His8-MBP-TEV-SalI,NotI-stop | SalI-NotI | His8-MBP-TEV | pCold for | pCold rev | Amp |
|
|
V114 | pColdII | NdeI-MBP-TEV-SalI,NotI-stop | SalI-NotI | MBP-TEV | pCold for | pCold rev | Amp |
|
|
V115 | pColdII | NdeI-XhoI-His6-stop | unknown | none | pCold for | pCold rev | Amp |
|
|
V116 | pET30a | NdeI-B562-XhoI-His6-stop | unknown | B562 | T7 Promoter | T7 Terminator | Kan |
|
|
V117 | pCold |
|
|
|
|
|
|
|
|
V118 | pGADzaA |
|
|
|
|
|
|
|
|
V119 | FBQH | His8-TEV-BamHI-XhoI | BamHI-XhoI | His8-TEV | FBQ-F | FBQ-R | Amp |
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V120 | FBQH | GST-TEV-BamHI-Xho I | BamHI-XhoI | GST-TEV | FBQ-F | FBQ-R | Amp |
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V121 | FBQH | His-MBP-TEV-BamHI-XhoI | BamHI-XhoI | His-MBP-TEV | FBQ-F | FBQ-R | Amp |
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V122 | PET30a | NdeI- His8- GB1 -Sal I -Not I-stop | Sal I-Not I | His8- GB1 | T7-P | T7-T | Kan | XY |
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V123 | pCDNA3.1 | CMV-T7-FLAG-MCS-Stop | XhoI-BamH I-EcoR I-Not I | FLAG | T7/CMV | BGH-R | Amp | XY |
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V124 | pCDNA3.1 | CMV-T7-HA-MCS-Stop | XhoI-BamH I-EcoR I-Not I | HA | T7/CMV | BGH-R | Amp | XY |
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V125 | pCDNA3.1 | CMV-T7-Strep-MCS-Stop | XhoI-BamH I-EcoR I-Not I | Strep | T7/CMV | BGH-R | Amp | XY |
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V126 | pCDNA3.1 | CMV-T7-SP-3XFLAG-MCS-Stop | XhoI-BamH I-EcoR I-Not I | 3XFLAG | T7/CMV | BGH-R | Amp | XY | SP=signal peptide |
V127 | pCDNA3.1 | CMV-T7-SP-FLAG-Clover-MCS-Stop | XhoI-BamH I-EcoR I-Not I | Clover/FLAG | T7/CMV | BGH-R | Amp | XY | SP=signal peptide |
V128 | pCDNA3.1 | CMV-T7-SP-HA-MCS-Stop | XhoI-BamH I-EcoR I-Not I | HA | T7/CMV | BGH-R | Amp | XY | SP=signal peptide |
V129 | pCDNA3.1 | CMV-T7-SP-Strep-mRubby2-MCS-Stop | XhoI-BamH I-EcoR I-Not I | Strep/mRubby2 | T7/CMV | BGH-R | Amp | XY | SP=signal peptide |
V130 | pCDNA3.1 | CMV-T7-SP-MCS-Stop | XhoI-BamH I-EcoR I-Not I | Strep | T7/CMV | BGH-R | Amp | XY |
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V131 | pCDNA3.1 | CMV-T7-Strep-mRubby2-MCS-Stop | XhoI-BamH I-EcoR I-Not I | Strep/mRubby2 | T7/CMV |