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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：
pREP10
- 库存：
大量
- 供应商：
钰博生物
- 规格：
1ug/5ug
pREP10
载体基本信息
| 出品公司: | Ybscience |
|---|---|
| 载体名称: | pREP10 |
| 质粒类型: | 哺乳动物表达载体；附加体载体 |
| 高拷贝/低拷贝: | 高拷贝 |
| 克隆方法: | 限制性内切酶，多克隆位点 |
| 启动子: | RSV |
| 载体大小: | 9287bp |
| 5' 测序引物及序列: | pREP Forward 5’ GCTCGATACAATAAACGCC 3’ |
| 3' 测序引物及序列: | SV40pA rev 5'GAAATTTGTGATGCTATTGC' |
| 载体标签: | -- |
| 载体抗性: | 氨苄青霉素 |
| 筛选标记: | Hygromycin |
| 克隆菌株: | DH5α 等 |
| 宿主细胞（系）: | 人类细胞；灵长类细胞；犬类细胞 |
| 备注: | pREP7载体是附加体型载体，可以在基因组外自主复制，类似的载体有： pREP4 PCEP4 pREP7 pREP10 pEBNA-DEST pEBVHisA pEBVHisB pEBVHisC pREP4/CAT pEBVHis-lacZ |
| 产品目录号: | -- |
| 稳定性: | 稳表达 |
| 组成型/诱导型: | 组成型 |
| 病毒/非病毒: | 非病毒 |
载体质粒图谱和多克隆位点信息
载体简介
载体序列
LOCUS Exported 9287 bp ds-DNA circular SYN 12-AUG-2016
DEFINITION synthetic circular DNA
KEYWORDS pREP10
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9287)
AUTHORS Biofeng Lab.
FEATURES Location/Qualifiers
source 1..9287
/organism="synthetic DNA construct"
/mol_type="other DNA"
polyA_signal 275..409
/note="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
promoter complement(476..737)
/note="RSV promoter"
/note="Rous sarcoma virus enhancer/promoter"
polyA_signal 1503..1551
/note="HSV TK poly(A) signal"
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
CDS complement(1593..2630)
/codon_start=1
/product="hygromycin B phosphotransferase"
/note="HygR"
/note="confers resistance to hygromycin"
/translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPDRE
MGEAN"
promoter complement(2673..2818)
/note="HSV TK promoter"
/note="herpes simplex virus thymidine kinase promoter"
rep_origin complement(3249..3837)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(4008..4868)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/note="AmpR"
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(4869..4973)
/gene="bla"
/note="AmpR promoter"
rep_origin 7049..8837
/note="oriP"
/note="Epstein-Barr virus oriP replication origin (Yates et
al., 2000)"
ORIGIN
1 tctagagtcg acctcatggc tgcgccccga cacccgccaa cacccgctga cgcgccctga
61 cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc
121 atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga ggcagccgga tcataatcag
181 ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa
241 cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg
301 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
361 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggatcg gtaccagctg
421 aagcttgcta gcggccgctc gaggccggca aggccggatc cgtacccagc ttggaggtgc
481 acaccaatgt ggtgaatggt caaatggcgt ttattgtatc gagctaggca cttaaataca
541 atatctctgc aatgcggaat tcagtggttc gtccaatcca tgtcagaccc gtctgttgcc
601 ttcctaataa ggcacgatcg taccacctta cttccaccaa tcggcatgca cggtgctttt
661 tctctccttg taaggcatgt tgctaactca tcgttaccat gttgcaagac tacaagagta
721 ttgcataaga ctacatttcc ccctccctat gcaaaagcga aactactata tcctgagggg
781 actcctaacc gcgtacaacc gaagccccgc ttttcgccta aacacaccct agtcccctca
841 gatacgcgta tatctggccc gtacatcgcg aagcagcgca aaacgcctaa ccctaagcag
901 attcttcatg caattgtcgg tcaagccttg ccttgttgta gcttaaattt tgctcgcgca
961 ctactcagcg acctccaaca cacaagcagg gagcagatac tggcttaact atgcggcatc
1021 agagcagatt gtactgagag tgcaccatac ggatctgcga tgataagctg tcaaacatga
1081 gaattggtcg actagcttgg cacgccagaa atccgcgcgg tggtttttgg gggtcggggg
1141 tgtttggcag ccacagacgc ccggtgttcg tgtcgcgcca gtacatgcgg tccatgccca
1201 ggccatccaa aaaccatggg tctgtctgct cagtccagtc gtggacctga ccccacgcaa
1261 cgcccaaaat aataaccccc acgaaccata aaccattccc catgggggac cccgtcccta
1321 acccacgggg ccagtggcta tggcagggcc tgccgccccg acgttggctg cgagccctgg
1381 gccttcaccc gaacttgggg ggtggggtgg ggaaaaggaa gaaacgcggg cgtattggcc
1441 ccaatggggt ctcggtgggg tatcgacaga gtgccagccc tgggaccgaa ccccgcgttt
1501 atgaacaaac gacccaacac ccgtgcgttt tattctgtct ttttattgcc gtcatagcgc
1561 gggttccttc cggtattgtc tccttccgtg tttcagttag cctcccccat ctcccgatcc
1621 ggacgagtgc tggggcgtcg gtttccacta tcggcgagta cttctacaca gccatcggtc
1681 cagacggccg cgcttctgcg ggcgatttgt gtacgcccga cagtcccggc tccggatcgg
1741 acgattgcgt cgcatcgacc ctgcgcccaa gctgcatcat cgaaattgcc gtcaaccaag
1801 ctctgataga gttggtcaag accaatgcgg agcatatacg cccggagccg cggcgatcct
1861 gcaagctccg gatgcctccg ctcgaagtag cgcgtctgct gctccataca agccaaccac
1921 ggcctccaga agaagatgtt ggcgacctcg tattgggaat ccccgaacat cgcctcgctc
1981 cagtcaatga ccgctgttat gcggccattg tccgtcagga cattgttgga gccgaaatcc
2041 gcgtgcacga ggtgccggac ttcggggcag tcctcggccc aaagcatcag ctcatcgaga
2101 gcctgcgcga cggacgcact gacggtgtcg tccatcacag tttgccagtg atacacatgg
2161 ggatcagcaa tcgcgcatat gaaatcacgc catgtagtgt attgaccgat tccttgcggt
2221 ccgaatgggc cgaacccgct cgtctggcta agatcggccg cagcgatcgc atccatggcc
2281 tccgcgaccg gctgcagaac agcgggcagt tcggtttcag gcaggtcttg caacgtgaca
2341 ccctgtgcac ggcgggagat gcaataggtc aggctctcgc tgaattcccc aatgtcaagc
2401 acttccggaa tcgggagcgc ggccgatgca aagtgccgat aaacataacg atctttgtag
2461 aaaccatcgg cgcagctatt tacccgcagg acatatccac gccctcctac atcgaagctg
2521 aaagcacgag attcttcgcc ctccgagagc tgcatcaggt cggagacgct gtcgaacttt
2581 tcgatcagaa acttctcgac agacgtcgcg gtgagttcag gctttttcat atctcattgc
2641 cccccgggat ctgcggcacg ctgttgacgc tgttaagcgg gtcgctgcag ggtcgctcgg
2701 tgttcgaggc cacacgcgtc accttaatat gcgaagtgga cctcggaccg cgccgccccg
2761 actgcatctg cgtgttcgaa ttcgccaatg acaagacgct gggcggggtt tgtgtcatca
2821 tagaactaaa gacatgcaaa tatatttctt ccggggacac cgccagcaaa cgcgagcaac
2881 gggccacggg gatgaagcag ggcggcacct cgctaacgga ttcaccactc caagaattgg
2941 agccaatcaa ttcttgcgga gaactgtgaa tgcgcaaacc aacccttggc agaacatatc
3001 catcgcgtcc gccatctcca gcagccgcac gcggcgcatc tcggggccga cgcgctgggc
3061 tacgtcttgc tggcgttcgc gacgcgaggc tggatggcct tccccattat gattcttctc
3121 gcttccggcg gcatcgggat gcccgcgttg caggccatgc tgtccaggca ggtagatgac
3181 gaccatcagg gacagcttca aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
3241 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
3301 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
3361 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
3421 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
3481 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
3541 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
3601 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
3661 tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag
3721 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
3781 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
3841 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
3901 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
3961 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
4021 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
4081 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
4141 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
4201 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
4261 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
4321 gctgcaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
4381 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
4441 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
4501 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
4561 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
4621 tcaacacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
4681 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
4741 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
4801 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
4861 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
4921 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
4981 ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa
5041 aataggcgta tcacgaggcc ctttcgtctt caagaattct catgtttgac agcttatcat
5101 cgataagctg atcctcacag gccgcaccca gcttttcttc cgttgcccca gtagcatctc
5161 tgtctggtga ccttgaagag gaagaggagg ggtcccgaga atccccatcc ctaccgtcca
5221 gcaaaaaggg ggacgaggaa tttgaggcct ggcttgaggc tcaggacgca aatcttgagg
5281 atgttcagcg ggagttttcc gggctgcgag taattggtga tgaggacgag gatggttcgg
5341 aggatgggga attttcagac ctggatctgt ctgacagcga ccatgaaggg gatgagggtg
5401 ggggggctgt tggagggggc aggagtctgc actccctgta ttcactgagc gtcgtctaat
5461 aaagatgtct attgatctct tttagtgtga atcatgtctg acgaggggcc aggtacagga
5521 cctggaaatg gcctaggaga gaagggagac acatctggac cagaaggctc cggcggcagt
5581 ggacctcaaa gaagaggggg tgataaccat ggacgaggac ggggaagagg acgaggacga
5641 ggaggcggaa gaccaggagc cccgggcggc tcaggatcag ggccaagaca tagagatggt
5701 gtccggagac cccaaaaacg tccaagttgc attggctgca aagggaccca cggtggaaca
5761 ggagcaggag caggagcggg aggggcagga gcaggaggtg gaggccgggg tcgaggaggc
5821 agtggaggcc ggggtcgagg aggtagtgga ggccggggtc gaggaggtag tggaggccgc
5881 cggggtagag gacgtgaaag agccaggggg ggaagtcgtg aaagagccag ggggagaggt
5941 cgtggacgtg gagaaaagag gcccaggagt cccagtagtc agtcatcatc atccgggtct
6001 ccaccgcgca ggccccctcc aggtagaagg ccatttttcc accctgtagg ggaagccgat
6061 tattttgaat accaccaaga aggtggccca gatggtgagc ctgacgtgcc cccgggagcg
6121 atagagcagg gccccgcaga tgacccagga gaaggcccaa gcactggacc ccggggtcag
6181 ggtgatggag gcaggcgcaa aaaaggaggg tggtttggaa agcatcgtgg tcaaggaggt
6241 tccaacccga aatttgagaa cattgcagaa ggtttaagag ctctcctggc taggagtcac
6301 gtagaaagga ctaccgacga aggaacttgg gtcgccggtg tgttcgtata tggaggtagt
6361 aagacctccc tttacaacct aaggcgagga actgcccttg ctattccaca atgtcgtctt
6421 acaccattga gtcgtctccc ctttggaatg gcccctggac ccggcccaca acctggcccg
6481 ctaagggagt ccattgtctg ttatttcatg gtctttttac aaactcatat atttgctgag
6541 gttttgaagg atgcgattaa ggaccttgtt atgacaaagc ccgctcctac ctgcaatatc
6601 agggtgactg tgtgcagctt tgacgatgga gtagatttgc ctccctggtt tccacctatg
6661 gtggaagggg ctgccgcgga gggtgatgac ggagatgacg gagatgaagg aggtgatgga
6721 gatgagggtg aggaagggca ggagtgatgt aacttgttag gagacgccct caatcgtatt
6781 aaaagccgtg tattcccccg cactaaagaa taaatcccca gtagacatca tgcgtgctgt
6841 tggtgtattt ctggccatct gtcttgtcac cattttcgtc ctcccaacat ggggcaattg
6901 ggcataccca tgttgtcacg tcactcagct ccgcgctcaa caccttctcg cgttggaaaa
6961 cattagcgac atttacctgg tgagcaatca gacatgcgac ggctttagcc tggcctcctt
7021 aaattcacct aagaatggga gcaaccagca ggaaaaggac aagcagcgaa aattcacgcc
7081 cccttgggag gtggcggcat atgcaaagga tagcactccc actctactac tgggtatcat
7141 atgctgactg tatatgcatg aggatagcat atgctacccg gatacagatt aggatagcat
7201 atactaccca gatatagatt aggatagcat atgctaccca gatatagatt aggatagcct
7261 atgctaccca gatataaatt aggatagcat atactaccca gatatagatt aggatagcat
7321 atgctaccca gatatagatt aggatagcct atgctaccca gatatagatt aggatagcat
7381 atgctaccca gatatagatt aggatagcat atgctatcca gatatttggg tagtatatgc
7441 tacccagata taaattagga tagcatatac taccctaatc tctattagga tagcatatgc
7501 tacccggata cagattagga tagcatatac tacccagata tagattagga tagcatatgc
7561 tacccagata tagattagga tagcctatgc tacccagata taaattagga tagcatatac
7621 tacccagata tagattagga tagcatatgc tacccagata tagattagga tagcctatgc
7681 tacccagata tagattagga tagcatatgc tatccagata tttgggtagt atatgctacc
7741 catggcaaca ttagcccacc gtgctctcag cgacctcgtg aatatgagga ccaacaaccc
7801 tgtgcttggc gctcaggcgc aagtgtgtgt aatttgtcct ccagatcgca gcaatcgcgc
7861 ccctatcttg gcccgcccac ctacttatgc aggtattccc cggggtgcca ttagtggttt
7921 tgtgggcaag tggtttgacc gcagtggtta gcggggttac aatcagccaa gttattacac
7981 ccttatttta cagtccaaaa ccgcagggcg gcgtgtgggg gctgacgcgt gcccccactc
8041 cacaatttca aaaaaaagag tggccacttg tctttgttta tgggccccat tggcgtggag
8101 ccccgtttaa ttttcggggg tgttagagac aaccagtgga gtccgctgct gtcggcgtcc
8161 actctctttc cccttgttac aaatagagtg taacaacatg gttcacctgt cttggtccct
8221 gcctgggaca catcttaata accccagtat catattgcac taggattatg tgttgcccat
8281 agccataaat tcgtgtgaga tggacatcca gtctttacgg cttgtcccca ccccatggat
8341 ttctattgtt aaagatattc agaatgtttc attcctacac tagtatttat tgcccaaggg
8401 gtttgtgagg gttatattgg tgtcatagca caatgccacc actgaacccc ccgtccaaat
8461 tttattctgg gggcgtcacc tgaaaccttg ttttcgagca cctcacatac accttactgt
8521 tcacaactca gcagttattc tattagctaa acgaaggaga atgaagaagc aggcgaagat
8581 tcaggagagt tcactgcccg ctccttgatc ttcagccact gcccttgtga ctaaaatggt
8641 tcactaccct cgtggaatcc tgaccccatg taaataaaac cgtgacagct catggggtgg
8701 gagatatcgc tgttccttag gaccctttta ctaaccctaa ttcgatagca tatgcttccc
8761 ttgggtaaca tatgctattg aattagggtt agtctggata gtatatacta ctacccggga
8821 agcatatgct acccgtttag ggttaacaag ggggccttat aaacactatt gctaatgccc
8881 tcttgagggt ccgcttatcg gtagctacac aggcccctct gattgacgtt ggtgtagcct
8941 cccgtagtct tcctgggccc ctgggaggta catgtccccc agcattggtg taagagcttc
9001 agccaagagt tacacataaa ggcaatgttg tgttgcagtc cacagactgc aaagtctgct
9061 ccaggatgaa agccactcag tgttggcaaa tgtgcacatc catttataag gatgtcaact
9121 acagtcagag aacccctttg tgtttggtcc ccccccgtgt cacatgtgga acagggccca
9181 gttggcaagt tgtaccaacc aactgaaggg attacatgca ctgccccgcg aagaaggggc
9241 agagatgtcg tagtcaggtt tagttcgtcc ggggcgggga tcgatcc
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文献和实验相关实验
Yale RNA prep (Rebecca D. Burdine, Michael J. Stern [modified by S.M. Hettenbach]) Add 8 ml of TRIZOL to 2 ml packed worms in 15 ml centrifuge tube. Vortex and invert tube to solubilize and lyse worms for at least 10 min.
，别人可以，自己确实不一定可以。 07 实验室的补助要涨了事实上，补助不降已经是学校和实验室对你最大的温柔。 08 等我搞科研发财了送外卖发财可能都比搞科研发财容易啊，发财了能怎么样，还不是做着用暴富的钱买进口试剂的梦。 09 我是老板带过的最有科研天赋的学生之一是的，每个人都会觉得他是其中之一，理论上，这样说没毛病。 10 师兄要发论文了，我帮过忙，能共一吧老板：共同通讯要不要啊，少年～～能坚持看完这篇推送的你，梦醒了吗？
A quick RNA mini-prep for Neurospora mycelial cultures
-prep RNA probed with ccg-1 DNA fragment. Total RNA from a series of transformants into bd A and frq7 A was examined for the presence of the ccg-1 gene transcript (Loros et al. 1989 Science 243:385-388). Each lane contains 10 µg of RNA (1/200
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pREP10
¥800 - 1200
