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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:
pDONR Zeo
- 库存:
大量
- 供应商:
钰博生物
- 规格:
1ug/5ug
pDONRZeo载体基本信息
| 出品公司: | Ybscience |
|---|---|
| 载体名称: | pDONR/Zeo, pDONRzeo, pDONR-Zeo |
| 质粒类型: | Gateway载体 |
| 高拷贝/低拷贝: | 高拷贝 |
| 启动子: | 无 |
| 克隆方法: | Gateway; BP反应 |
| 载体大小: | 4291bp |
| 5' 测序引物及序列: | M13F: TGTAAAACGACGGCCAGT |
| 3' 测序引物及序列: | M13R: CAGGAAACAGCTATGAC |
| 载体标签: | 无 |
| 载体抗性: | 博来霉素、氯霉素 |
| 备注: | pDONR/zeo载体本身含有CCDb致死基因,能够使得普通的DH5alpha等细菌死亡,必须使用DB3.1感受态细胞进行质粒扩增; 构建Gateway系统入门克隆时需要使用ccdB敏感的 OmniMAX2 感受态细胞。 |
| 病毒/非病毒: | 非病毒 |
pDONRZeo载体质粒图谱和多克隆位点信息


pDONRZeo载体简介
pDONRZeo载体序列
LOCUS pDONR\Zeo 4291 bp DNA SYN
DEFINITION pDONR\Zeo
ACCESSION
KEYWORDS
SOURCE
ORGANISM other sequences; artificial sequences; vectors.
COMMENT This file is created by Vector NTI
http://www.biofeng.com/
COMMENT VNTAUTHORNAME|biofeng.com|
FEATURES Location/Qualifiers
source 1..4291
/organism="pDONR/Zeo"
/mol_type="other DNA"
terminator complement(268..295)
/label="rrnB_T2_terminator"
terminator complement(427..470)
/label="rrnB_T1_terminator"
misc_feature 522..544
/label="M13_pUC_fwd_primer"
promoter 537..553
/label="M13_forward20_primer"
misc_feature complement(569..652)
/label="attL1"
misc_feature 570..801
/label="attP"
misc_feature 656..768
/label="attR1"
misc_feature complement(668..768)
/label="attR2"
misc_feature complement(1197..1502)
/label="ccdB"
gene complement(1847..2506)
/label="CAT/CamR"
/gene="CAT/CamR"
CDS complement(1847..2506)
/label="ORF frame 1"
misc_feature 2787..2887
/label="attR2"
promoter complement(3004..3022)
/label="T7_promoter"
promoter complement(3023..3041)
/label="M13_reverse_primer"
misc_feature complement(3111..3485)
/label="sh_ble"
gene complement(3114..3485)
/label="bleo"
/gene="bleo"
promoter complement(3486..3553)
/label="EM7_promoter"
rep_origin 3625..4244
/label="pBR322_origin"
ORIGIN
1 CTTTCCTGCG TTATCCCCTG ATTCTGTGGA TAACCGTATT ACCGCCTTTG AGTGAGCTGA
61 TACCGCTCGC CGCAGCCGAA CGACCGAGCG CAGCGAGTCA GTGAGCGAGG AAGCGGAAGA
121 GCGCCCAATA CGCAAACCGC CTCTCCCCGC GCGTTGGCCG ATTCATTAAT GCAGCTGGCA
181 CGACAGGTTT CCCGACTGGA AAGCGGGCAG TGAGCGCAAC GCAATTAATA CGCGTACCGC
241 TAGCCAGGAA GAGTTTGTAG AAACGCAAAA AGGCCATCCG TCAGGATGGC CTTCTGCTTA
301 GTTTGATGCC TGGCAGTTTA TGGCGGGCGT CCTGCCCGCC ACCCTCCGGG CCGTTGCTTC
361 ACAACGTTCA AATCCGCTCC CGGCGGATTT GTCCTACTCA GGAGAGCGTT CACCGACAAA
421 CAACAGATAA AACGAAAGGC CCAGTCTTCC GACTGAGCCT TTCGTTTTAT TTGATGCCTG
481 GCAGTTCCCT ACTCTCGCGT TAACGCTAGC ATGGATGTTT TCCCAGTCAC GACGTTGTAA
541 AACGACGGCC AGTCTTAAGC TCGGGCCCCA AATAATGATT TTATTTTGAC TGATAGTGAC
601 CTGTTCGTTG CAACACATTG ATGAGCAATG CTTTTTTATA ATGCCAACTT TGTACAAAAA
661 AGCTGAACGA GAAACGTAAA ATGATATAAA TATCAATATA TTAAATTAGA TTTTGCATAA
721 AAAACAGACT ACATAATACT GTAAAACACA ACATATCCAG TCACTATGAA TCAACTACTT
781 AGATGGTATT AGTGACCTGT AGTCGACCGA CAGCCTTCCA AATGTTCTTC GGGTGATGCT
841 GCCAACTTAG TCGACCGACA GCCTTCCAAA TGTTCTTCTC AAACGGAATC GTCGTATCCA
901 GCCTACTCGC TATTGTCCTC AATGCCGTAT TAAATCATAA AAAGAAATAA GAAAAAGAGG
961 TGCGAGCCTC TTTTTTGTGT GACAAAATAA AAACATCTAC CTATTCATAT ACGCTAGTGT
1021 CATAGTCCTG AAAATCATCT GCATCAAGAA CAATTTCACA ACTCTTATAC TTTTCTCTTA
1081 CAAGTCGTTC GGCTTCATCT GGATTTTCAG CCTCTATACT TACTAAACGT GATAAAGTTT
1141 CTGTAATTTC TACTGTATCG ACCTGCAGAC TGGCTGTGTA TAAGGGAGCC TGACATTTAT
1201 ATTCCCCAGA ACATCAGGTT AATGGCGTTT TTGATGTCAT TTTCGCGGTG GCTGAGATCA
1261 GCCACTTCTT CCCCGATAAC GGAGACCGGC ACACTGGCCA TATCGGTGGT CATCATGCGC
1321 CAGCTTTCAT CCCCGATATG CACCACCGGG TAAAGTTCAC GGGAGACTTT ATCTGACAGC
1381 AGACGTGCAC TGGCCAGGGG GATCACCATC CGTCGCCCGG GCGTGTCAAT AATATCACTC
1441 TGTACATCCA CAAACAGACG ATAACGGCTC TCTCTTTTAT AGGTGTAAAC CTTAAACTGC
1501 ATTTCACCAG CCCCTGTTCT CGTCAGCAAA AGAGCCGTTC ATTTCAATAA ACCGGGCGAC
1561 CTCAGCCATC CCTTCCTGAT TTTCCGCTTT CCAGCGTTCG GCACGCAGAC GACGGGCTTC
1621 ATTCTGCATG GTTGTGCTTA CCAGACCGGA GATATTGACA TCATATATGC CTTGAGCAAC
1681 TGATAGCTGT CGCTGTCAAC TGTCACTGTA ATACGCTGCT TCATAGCATA CCTCTTTTTG
1741 ACATACTTCG GGTATACATA TCAGTATATA TTCTTATACC GCAAAAATCA GCGCGCAAAT
1801 ACGCATACTG TTATCTGGCT TTTAGTAAGC CGGATCCACG CGGCGTTTAC GCCCCGCCCT
1861 GCCACTCATC GCAGTACTGT TGTAATTCAT TAAGCATTCT GCCGACATGG AAGCCATCAC
1921 AGACGGCATG ATGAACCTGA ATCGCCAGCG GCATCAGCAC CTTGTCGCCT TGCGTATAAT
1981 ATTTGCCCAT GGTGAAAACG GGGGCGAAGA AGTTGTCCAT ATTGGCCACG TTTAAATCAA
2041 AACTGGTGAA ACTCACCCAG GGATTGGCTG AGACGAAAAA CATATTCTCA ATAAACCCTT
2101 TAGGGAAATA GGCCAGGTTT TCACCGTAAC ACGCCACATC TTGCGAATAT ATGTGTAGAA
2161 ACTGCCGGAA ATCGTCGTGG TATTCACTCC AGAGCGATGA AAACGTTTCA GTTTGCTCAT
2221 GGAAAACGGT GTAACAAGGG TGAACACTAT CCCATATCAC CAGCTCACCG TCTTTCATTG
2281 CCATACGGAA TTCCGGATGA GCATTCATCA GGCGGGCAAG AATGTGAATA AAGGCCGGAT
2341 AAAACTTGTG CTTATTTTTC TTTACGGTCT TTAAAAAGGC CGTAATATCC AGCTGAACGG
2401 TCTGGTTATA GGTACATTGA GCAACTGACT GAAATGCCTC AAAATGTTCT TTACGATGCC
2461 ATTGGGATAT ATCAACGGTG GTATATCCAG TGATTTTTTT CTCCATTTTA GCTTCCTTAG
2521 CTCCTGAAAA TCTCGATAAC TCAAAAAATA CGCCCGGTAG TGATCTTATT TCATTATGGT
2581 GAAAGTTGGA ACCTCTTACG TGCCGATCAA CGTCTCATTT TCGCCAAAAG TTGGCCCAGG
2641 GCTTCCCGGT ATCAACAGGG ACACCAGGAT TTATTTATTC TGCGAAGTGA TCTTCCGTCA
2701 CAGGTATTTA TTCGGCGCAA AGTGCGTCGG GTGATGCTGC CAACTTAGTC GACTACAGGT
2761 CACTAATACC ATCTAAGTAG TTGATTCATA GTGACTGGAT ATGTTGTGTT TTACAGTATT
2821 ATGTAGTCTG TTTTTTATGC AAAATCTAAT TTAATATATT GATATTTATA TCATTTTACG
2881 TTTCTCGTTC AGCTTTCTTG TACAAAGTTG GCATTATAAG AAAGCATTGC TTATCAATTT
2941 GTTGCAACGA ACAGGTCACT ATCAGTCAAA ATAAAATCAT TATTTGCCAT CCAGCTGATA
3001 TCCCCTATAG TGAGTCGTAT TACATGGTCA TAGCTGTTTC CTGGCAGCTC TGGCCCGTGT
3061 CTCAAAATCT CTGATGTTAC ATTGCACAAG ATAAAATAAT ATCATCATGA TCAGTCCTGC
3121 TCCTCGGCCA CGAAGTGCAC GCAGTTGCCG GCCGGGTCGC GCAGGGCGAA CTCCCGCCCC
3181 CACGGCTGCT CGCCGATCTC GGTCATGGCC GGCCCGGAGG CGTCCCGGAA GTTCGTGGAC
3241 ACGACCTCCG ACCACTCGGC GTACAGCTCG TCCAGGCCGC GCACCCACAC CCAGGCCAGG
3301 GTGTTGTCCG GCACCACCTG GTCCTGGACC GCGCTGATGA ACAGGGTCAC GTCGTCCCGG
3361 ACCACACCGG CGAAGTCGTC CTCCACGAAG TCCCGGGAGA ACCCGAGCCG GTCGGTCCAG
3421 AACTCGACCG CTCCGGCGAC GTCGCGCGCG GTGAGCACCG GAACGGCACT GGTCAACTTG
3481 GCCATGGTTT AGTTCCTCAC CTTGTCGTAT TATACTATGC CGATATACTA TGCCGATGAT
3541 TAATTGTCAA CACGTGCTGA TCATGACCAA AATCCCTTAA CGTGAGTTAC GCGTCGTTCC
3601 ACTGAGCGTC AGACCCCGTA GAAAAGATCA AAGGATCTTC TTGAGATCCT TTTTTTCTGC
3661 GCGTAATCTG CTGCTTGCAA ACAAAAAAAC CACCGCTACC AGCGGTGGTT TGTTTGCCGG
3721 ATCAAGAGCT ACCAACTCTT TTTCCGAAGG TAACTGGCTT CAGCAGAGCG CAGATACCAA
3781 ATACTGTTCT TCTAGTGTAG CCGTAGTTAG GCCACCACTT CAAGAACTCT GTAGCACCGC
3841 CTACATACCT CGCTCTGCTA ATCCTGTTAC CAGTGGCTGC TGCCAGTGGC GATAAGTCGT
3901 GTCTTACCGG GTTGGACTCA AGACGATAGT TACCGGATAA GGCGCAGCGG TCGGGCTGAA
3961 CGGGGGGTTC GTGCACACAG CCCAGCTTGG AGCGAACGAC CTACACCGAA CTGAGATACC
4021 TACAGCGTGA GCTATGAGAA AGCGCCACGC TTCCCGAAGG GAGAAAGGCG GACAGGTATC
4081 CGGTAAGCGG CAGGGTCGGA ACAGGAGAGC GCACGAGGGA GCTTCCAGGG GGAAACGCCT
4141 GGTATCTTTA TAGTCCTGTC GGGTTTCGCC ACCTCTGACT TGAGCGTCGA TTTTTGTGAT
4201 GCTCGTCAGG GGGGCGGAGC CTATGGAAAA ACGCCAGCAA CGCGGCCTTT TTACGGTTCC
4261 TGGCCTTTTG CTGGCCTTTT GCTCACATGT T
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文献和实验Replication timing by comparative hybridization
% unbudded), remove 20 ml for the "Uninduced" control, and add dry galactose (2% w/v final concentration) to the remainder of the culture. R-KT-DIR ) or 4 hr ( R-Zeo-DIR ), then add dry glucose (2% w/v final concentration). Thirty minutes later, shift
用传统的限制酶切和连接的方法产生入门克隆。这些载体配合合适的目的载体,可以用于表达天然蛋白或带有N端或C端融合标签的重组蛋白。为了在真核细胞中有效地翻译蛋白,所有的5个Gateway入门载体提供了Kozak 序列。此外,pENTR11提供了SD (Shine-Dalgarno)序列,便于在大肠杆菌中有效的翻译。2、PCR重组克隆 重组是从PCR产物创建Gateway入门克隆的另一种方法。这种方法是通过合并attB位点到上游和下游引物上,然后共同孵育PCR扩增产物和pDONR载体(包含attP位点
Replication timing by comparative hybridization
% unbudded), remove 20 ml for the "Uninduced" control, and add dry galactose (2% w/v final concentration) to the remainder of the culture. R-KT-DIR ) or 4 hr ( R-Zeo-DIR ), then add dry glucose (2% w/v final concentration). Thirty minutes later, shift
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