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- 详细信息
- 技术资料
- 保存条件：
-20
- 保质期：
2年
- 英文名：
Plasmid#18982;pHIV-H2B-mRFP
- 库存：
100
- 供应商：
上海烜雅生物科技有限公司
- 规格：
干粉/液体
基本信息
名称：pHIV-H2BmRFP慢病毒表达质粒
别称: Plasmid#18982;pHIV-H2B-mRFP
质粒属性
质粒简介
PHIV-H2BmRFP background vector HIV-Zsgreen, which is a high copy plasmid, is a slow virus vector, in the transformation of the cloned strain is Stbl3, at 37 C. The culture of the plasmid containing ampicillin.
质粒图谱
质粒序列
LOCUS Exported 8045 bp ds-DNA circular SYN 12-JUL-2018
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pHIV-H2BmRFP
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8045)
AUTHORS Triple Threat
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..8045
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 238..617
/note="CMV enhancer"
/note="human cytomegalovirus immediate early enhancer"
promoter 619..817
/note="CMV promoter"
/note="human cytomegalovirus (CMV) immediate early
promoter"
LTR 835..1015
/note="5' LTR (truncated)"
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 1062..1187
/note="HIV-1 Psi"
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1686..1919
/note="RRE"
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
misc_feature 2446..2563
/note="cPPT/CTS"
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 2627..3805
/note="EF-1-alpha promoter"
/note="strong constitutive promoter for human elongation
factor EF-1-alpha"
misc_feature 3851..4437
/note="IRES2"
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
misc_feature 4438..4818
/note="H2B"
CDS 4837..5514
/codon_start=1
/product="monomeric derivative of DsRed (Campbell et al.,
2002)"
/note="mRFP1"
/note="mammalian codon-optimized"
/translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
TGA"
LTR 6051..6231
/note="5' LTR (truncated)"
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
rep_origin complement(6293..6881)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(7052..7912)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/note="AmpR"
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(7913..8017)
/gene="bla"
/note="AmpR promoter"
ORIGIN
1 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
61 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
121 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
181 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac
241 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
301 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
361 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
421 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
481 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
541 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
601 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
661 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
721 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
781 gcggtaggcg tgtacggtgg gaggtctata taagcagcgc gttttgcctg tactgggtct
841 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
901 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
961 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc
1021 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc
1081 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa
1141 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg
1201 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata
1261 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc
1321 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga
1381 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc
1441 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca
1501 aaagtaagac caccgcacag caagcggccg gccgcgctga tcttcagacc tggaggagga
1561 gatatgaggg acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca
1621 ttaggagtag cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg
1681 ggaataggag ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcg
1741 tcaatgacgc tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac
1801 aatttgctga gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc
1861 aagcagctcc aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg
1921 gggatttggg gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt
1981 tggagtaata aatctctgga acagatttgg aatcacacga cctggatgga gtgggacaga
2041 gaaattaaca attacacaag cttaatacac tccttaattg aagaatcgca aaaccagcaa
2101 gaaaagaatg aacaagaatt attggaatta gataaatggg caagtttgtg gaattggttt
2161 aacataacaa attggctgtg gtatataaaa ttattcataa tgatagtagg aggcttggta
2221 ggtttaagaa tagtttttgc tgtactttct atagtgaata gagttaggca gggatattca
2281 ccattatcgt ttcagaccca cctcccaacc ccgaggggac ccgacaggcc cgaaggaata
2341 gaagaagaag gtggagagag agacagagac agatccattc gattagtgaa cggatcggca
2401 ctgcgtgcgc caattctgca gacaaatggc agtattcatc cacaatttta aaagaaaagg
2461 ggggattggg gggtacagtg caggggaaag aatagtagac ataatagcaa cagacataca
2521 aactaaagaa ttacaaaaac aaattacaaa aattcaaaat tttcgggttt attacaggga
2581 cagcagagat ccagtttggt tagtaccggg cccgctctag ccgtgaggct ccggtgcccg
2641 tcagtgggca gagcgcacat cgcccacagt ccccgagaag ttggggggag gggtcggcaa
2701 ttgaaccggt gcctagagaa ggtggcgcgg ggtaaactgg gaaagtgatg tcgtgtactg
2761 gctccgcctt tttcccgagg gtgggggaga accgtatata agtgcagtag tcgccgtgaa
2821 cgttcttttt cgcaacgggt ttgccgccag aacacaggta agtgccgtgt gtggttcccg
2881 cgggcctggc ctctttacgg gttatggccc ttgcgtgcct tgaattactt ccacctggct
2941 gcagtacgtg attcttgatc ccgagcttcg ggttggaagt gggtgggaga gttcgaggcc
3001 ttgcgcttaa ggagcccctt cgcctcgtgc ttgagttgag gcctggcctg ggcgctgggg
3061 ccgccgcgtg cgaatctggt ggcaccttcg cgcctgtctc gctgctttcg ataagtctct
3121 agccatttaa aatttttgat gacctgctgc gacgcttttt ttctggcaag atagtcttgt
3181 aaatgcgggc caagatctgc acactggtat ttcggttttt ggggccgcgg gcggcgacgg
3241 ggcccgtgcg tcccagcgca catgttcggc gaggcggggc ctgcgagcgc ggccaccgag
3301 aatcggacgg gggtagtctc aagctggccg gcctgctctg gtgcctggcc tcgcgccgcc
3361 gtgtatcgcc ccgccctggg cggcaaggct ggcccggtcg gcaccagttg cgtgagcgga
3421 aagatggccg cttcccggcc ctgctgcagg gagctcaaaa tggaggacgc ggcgctcggg
3481 agagcgggcg ggtgagtcac ccacacaaag gaaaagggcc tttccgtcct cagccgtcgc
3541 ttcatgtgac tccacggagt accgggcgcc gtccaggcac ctcgattagt tctcgagctt
3601 ttggagtacg tcgtctttag gttgggggga ggggttttat gcgatggagt ttccccacac
3661 tgagtgggtg gagactgaag ttaggccagc ttggcacttg atgtaattct ccttggaatt
3721 tgcccttttt gagtttggat cttggttcat tctcaagcct cagacagtgg ttcaaagttt
3781 ttttcttcca tttcaggtgt cgtgagcggc cgctgagtta actattctag agtacccggg
3841 ctaggatccg cccctctccc tccccccccc ctaacgttac tggccgaagc cgcttggaat
3901 aaggccggtg tgcgtttgtc tatatgttat tttccaccat attgccgtct tttggcaatg
3961 tgagggcccg gaaacctggc cctgtcttct tgacgagcat tcctaggggt ctttcccctc
4021 tcgccaaagg aatgcaaggt ctgttgaatg tcgtgaagga agcagttcct ctggaagctt
4081 cttgaagaca aacaacgtct gtagcgaccc tttgcaggca gcggaacccc ccacctggcg
4141 acaggtgcct ctgcggccaa aagccacgtg tataagatac acctgcaaag gcggcacaac
4201 cccagtgcca cgttgtgagt tggatagttg tggaaagagt caaatggctc tcctcaagcg
4261 tattcaacaa ggggctgaag gatgcccaga aggtacccca ttgtatggga tctgatctgg
4321 ggcctcggtg cacatgcttt acatgtgttt agtcgaggtt aaaaaaacgt ctaggccccc
4381 cgaaccacgg ggacgtggtt ttcctttgaa aaacacgatg ataatatggc cacaaccatg
4441 tcaccagagc cagcgaagtc tgctcccgcc ccgaaaaagg gctccaagaa ggcggtgact
4501 aaggcgcaga agaaaggcgg caagaagcgc aagcgcagcc gcaaggagag ctattccatc
4561 tatgtgtaca aggttctgaa gcaggtccac cctgacaccg gcatttcgtc caaggccatg
4621 ggcatcatga attcgtttgt gaacgacatt ttcgagcgca tcgcaggtga ggcttcccgc
4681 ctggcgcatt acaacaagcg ctcgaccatc acctccaggg agatccagac ggccgtgcgc
4741 ctgctgctgc ctggggagtt ggccaagcac gccgtgtccg agggtactaa ggccatcacc
4801 aagtacacca gcgctaagga tccaccggtc gccaccatgg cctcctccga ggacgtcatc
4861 aaggagttca tgcgcttcaa ggtgcgcatg gagggctccg tgaacggcca cgagttcgag
4921 atcgagggcg agggcgaggg ccgcccctac gagggcaccc agaccgccaa gctgaaggtg
4981 accaagggcg gccccctgcc cttcgcctgg gacatcctgt cccctcagtt ccagtacggc
5041 tccaaggcct acgtgaagca ccccgccgac atccccgact acttgaagct gtccttcccc
5101 gagggcttca agtgggagcg cgtgatgaac ttcgaggacg gcggcgtggt gaccgtgacc
5161 caggactcct ccctgcagga cggcgagttc atctacaagg tgaagctgcg cggcaccaac
5221 ttcccctccg acggccccgt aatgcagaag aagaccatgg gctgggaggc ctccaccgag
5281 cggatgtacc ccgaggacgg cgccctgaag ggcgagatca agatgaggct gaagctgaag
5341 gacggcggcc actacgacgc cgaggtcaag accacctaca tggccaagaa gcccgtgcag
5401 ctgcccggcg cctacaagac cgacatcaag ctggacatca cctcccacaa cgaggactac
5461 accatcgtgg aacagtacga gcgcgccgag ggccgccact ccaccggcgc ctaagcggcc
5521 gcatcgatac cgtcgacctc gatcgagacc tagaaaaaca tggagcaatc acaagtagca
5581 atacagcagc taccaatgct gattgtgcct ggctagaagc acaagaggag gaggaggtgg
5641 gttttccagt cacacctcag gtacctttaa gaccaatgac ttacaaggca gctgtagatc
5701 ttagccactt tttaaaagaa aaggggggac tggaagggct aattcactcc caacgaagac
5761 aagatatcct tgatctgtgg atctaccaca cacaaggcta cttccctgat tggcagaact
5821 acacaccagg gccagggatc agatatccac tgacctttgg atggtgctac aagctagtac
5881 cagttgagca agagaaggta gaagaagcca atgaaggaga gaacacccgc ttgttacacc
5941 ctgtgagcct gcatgggatg gatgacccgg agagagaagt attagagtgg aggtttgaca
6001 gccgcctagc atttcatcac atggcccgag agctgcatcc ggactgtact gggtctctct
6061 ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc
6121 ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg
6181 gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc agcatgtgag
6241 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
6301 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
6361 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
6421 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
6481 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
6541 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
6601 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
6661 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
6721 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
6781 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
6841 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
6901 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
6961 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
7021 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
7081 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
7141 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
7201 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
7261 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
7321 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
7381 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
7441 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
7501 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
7561 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
7621 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
7681 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
7741 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
7801 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
7861 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
7921 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
7981 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
8041 ctgac
//
质粒菌株产品操作说明书
一、扩增流程
收到产品后，请先根据产品管壁标签来判断产品形式，并在扩增前准确查找该质粒菌株的抗性、感受态和培养温度。
1、质粒干粉（常温运输，存于-20度，90天保质期，请务必转化挑单克隆培养，不要直接使用和测序）
①收到质粒干粉后请先5000rpm离心1min，再加入20μl ddH2O去离子水溶解质粒；
②取1支100μl 感受态于冰上解冻10min，加入2μl质粒，再冰浴30min后，42℃热激60s，不要搅动，再冰浴2min；（从第二步开始均要在超净工作台中无菌操作）
③加入900μl无抗的LB液体培养基，180rpm震荡37℃培养45min；
④6000rpm离心5min，仅留100μl上清液重悬细菌沉淀，并涂布至目标质粒抗性的LB平板上；（可使用本平台的平板涂布专用玻璃珠进行涂布，可以比传统涂布方法获得更多转化子）
⑤将平板正向培养1h，再倒置37℃培养14h。如果要求是30度则培养20h；
（菌落过多则将质粒稀释后再转化。没有菌落则加入10μl质粒转化。另不要直接转表达感受态，要先转克隆感受态，重提质粒后再导入表达感受态）
⑥挑取单菌落至LB液体培养基中，加入对应抗生素，220rpm震荡培养14h，根据实验需要和质粒提取试剂盒说明书提取质粒。
2、甘油菌种（冰袋运输，存于-80℃，保质期90天，请务必划线挑单克隆培养）
四区划线后挑单菌落培养，酵母菌需要先液体复苏再四区划线，再挑单菌落液体培养。
3、穿刺菌种（冰袋运输，存于4℃，保质期7天）
穿刺接种，液体培养后四区划线，再挑单菌落液体培养。
4、菌落平板（冰袋运输，存于4℃，保质期7天）
直接挑取单菌落至液体培养基中。
5、液体质粒（冰袋运输，存于-20℃，保质期90天）
单独提取的液体质粒收到后可直接使用。
6、滤纸质粒（常温运输，存于-20度，90天保质期，请务必转化挑单克隆培养，不要直接使用和测序）
收到货后将滤纸画圈部分剪下放入EP管中，加100ul无菌水将滤纸浸湿并浸泡5min，吸取5ul质粒转化，离心全涂。
二、转化图片
名称：pHIV-H2BmRFP慢病毒表达质粒
别称: Plasmid#18982;pHIV-H2B-mRFP
| 启动子: | EF1a |
|---|---|
| 复制子: | pUC |
| 质粒分类: | 病毒系列，慢病毒克隆载体 |
| 质粒大小: | 8045bp |
| 原核抗性: | Amp |
| 克隆菌株: | Stbl3 |
| 培养条件: | 37度 |
| 表达宿主: | 哺乳细胞 |
| 诱导方式: | 无须诱导，瞬时表达 |
| 5'测序引物: | CMV-F：CGCAAATGGGCGGTAGGCGTG |
| 3'测序引物: | 根据序列设计引物 |
质粒属性
| 载体宿主： | 哺乳细胞,慢病毒 |
|---|---|
| 载体用途： | 蛋白表达 |
| 基因种属： | 空载体 |
| 基因类型： | ORF |
| 原核抗性： | Amp |
| 筛选标记： | |
| 荧光蛋白： | 红色 |
质粒简介
PHIV-H2BmRFP background vector HIV-Zsgreen, which is a high copy plasmid, is a slow virus vector, in the transformation of the cloned strain is Stbl3, at 37 C. The culture of the plasmid containing ampicillin.
质粒图谱
质粒序列
LOCUS Exported 8045 bp ds-DNA circular SYN 12-JUL-2018
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pHIV-H2BmRFP
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8045)
AUTHORS Triple Threat
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..8045
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 238..617
/note="CMV enhancer"
/note="human cytomegalovirus immediate early enhancer"
promoter 619..817
/note="CMV promoter"
/note="human cytomegalovirus (CMV) immediate early
promoter"
LTR 835..1015
/note="5' LTR (truncated)"
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 1062..1187
/note="HIV-1 Psi"
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1686..1919
/note="RRE"
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
misc_feature 2446..2563
/note="cPPT/CTS"
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 2627..3805
/note="EF-1-alpha promoter"
/note="strong constitutive promoter for human elongation
factor EF-1-alpha"
misc_feature 3851..4437
/note="IRES2"
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
misc_feature 4438..4818
/note="H2B"
CDS 4837..5514
/codon_start=1
/product="monomeric derivative of DsRed (Campbell et al.,
2002)"
/note="mRFP1"
/note="mammalian codon-optimized"
/translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
TGA"
LTR 6051..6231
/note="5' LTR (truncated)"
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
rep_origin complement(6293..6881)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(7052..7912)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/note="AmpR"
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(7913..8017)
/gene="bla"
/note="AmpR promoter"
ORIGIN
1 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
61 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
121 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
181 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac
241 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
301 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
361 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
421 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
481 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
541 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
601 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
661 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
721 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
781 gcggtaggcg tgtacggtgg gaggtctata taagcagcgc gttttgcctg tactgggtct
841 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
901 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
961 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc
1021 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc
1081 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa
1141 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg
1201 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata
1261 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc
1321 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga
1381 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc
1441 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca
1501 aaagtaagac caccgcacag caagcggccg gccgcgctga tcttcagacc tggaggagga
1561 gatatgaggg acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca
1621 ttaggagtag cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg
1681 ggaataggag ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcg
1741 tcaatgacgc tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac
1801 aatttgctga gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc
1861 aagcagctcc aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg
1921 gggatttggg gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt
1981 tggagtaata aatctctgga acagatttgg aatcacacga cctggatgga gtgggacaga
2041 gaaattaaca attacacaag cttaatacac tccttaattg aagaatcgca aaaccagcaa
2101 gaaaagaatg aacaagaatt attggaatta gataaatggg caagtttgtg gaattggttt
2161 aacataacaa attggctgtg gtatataaaa ttattcataa tgatagtagg aggcttggta
2221 ggtttaagaa tagtttttgc tgtactttct atagtgaata gagttaggca gggatattca
2281 ccattatcgt ttcagaccca cctcccaacc ccgaggggac ccgacaggcc cgaaggaata
2341 gaagaagaag gtggagagag agacagagac agatccattc gattagtgaa cggatcggca
2401 ctgcgtgcgc caattctgca gacaaatggc agtattcatc cacaatttta aaagaaaagg
2461 ggggattggg gggtacagtg caggggaaag aatagtagac ataatagcaa cagacataca
2521 aactaaagaa ttacaaaaac aaattacaaa aattcaaaat tttcgggttt attacaggga
2581 cagcagagat ccagtttggt tagtaccggg cccgctctag ccgtgaggct ccggtgcccg
2641 tcagtgggca gagcgcacat cgcccacagt ccccgagaag ttggggggag gggtcggcaa
2701 ttgaaccggt gcctagagaa ggtggcgcgg ggtaaactgg gaaagtgatg tcgtgtactg
2761 gctccgcctt tttcccgagg gtgggggaga accgtatata agtgcagtag tcgccgtgaa
2821 cgttcttttt cgcaacgggt ttgccgccag aacacaggta agtgccgtgt gtggttcccg
2881 cgggcctggc ctctttacgg gttatggccc ttgcgtgcct tgaattactt ccacctggct
2941 gcagtacgtg attcttgatc ccgagcttcg ggttggaagt gggtgggaga gttcgaggcc
3001 ttgcgcttaa ggagcccctt cgcctcgtgc ttgagttgag gcctggcctg ggcgctgggg
3061 ccgccgcgtg cgaatctggt ggcaccttcg cgcctgtctc gctgctttcg ataagtctct
3121 agccatttaa aatttttgat gacctgctgc gacgcttttt ttctggcaag atagtcttgt
3181 aaatgcgggc caagatctgc acactggtat ttcggttttt ggggccgcgg gcggcgacgg
3241 ggcccgtgcg tcccagcgca catgttcggc gaggcggggc ctgcgagcgc ggccaccgag
3301 aatcggacgg gggtagtctc aagctggccg gcctgctctg gtgcctggcc tcgcgccgcc
3361 gtgtatcgcc ccgccctggg cggcaaggct ggcccggtcg gcaccagttg cgtgagcgga
3421 aagatggccg cttcccggcc ctgctgcagg gagctcaaaa tggaggacgc ggcgctcggg
3481 agagcgggcg ggtgagtcac ccacacaaag gaaaagggcc tttccgtcct cagccgtcgc
3541 ttcatgtgac tccacggagt accgggcgcc gtccaggcac ctcgattagt tctcgagctt
3601 ttggagtacg tcgtctttag gttgggggga ggggttttat gcgatggagt ttccccacac
3661 tgagtgggtg gagactgaag ttaggccagc ttggcacttg atgtaattct ccttggaatt
3721 tgcccttttt gagtttggat cttggttcat tctcaagcct cagacagtgg ttcaaagttt
3781 ttttcttcca tttcaggtgt cgtgagcggc cgctgagtta actattctag agtacccggg
3841 ctaggatccg cccctctccc tccccccccc ctaacgttac tggccgaagc cgcttggaat
3901 aaggccggtg tgcgtttgtc tatatgttat tttccaccat attgccgtct tttggcaatg
3961 tgagggcccg gaaacctggc cctgtcttct tgacgagcat tcctaggggt ctttcccctc
4021 tcgccaaagg aatgcaaggt ctgttgaatg tcgtgaagga agcagttcct ctggaagctt
4081 cttgaagaca aacaacgtct gtagcgaccc tttgcaggca gcggaacccc ccacctggcg
4141 acaggtgcct ctgcggccaa aagccacgtg tataagatac acctgcaaag gcggcacaac
4201 cccagtgcca cgttgtgagt tggatagttg tggaaagagt caaatggctc tcctcaagcg
4261 tattcaacaa ggggctgaag gatgcccaga aggtacccca ttgtatggga tctgatctgg
4321 ggcctcggtg cacatgcttt acatgtgttt agtcgaggtt aaaaaaacgt ctaggccccc
4381 cgaaccacgg ggacgtggtt ttcctttgaa aaacacgatg ataatatggc cacaaccatg
4441 tcaccagagc cagcgaagtc tgctcccgcc ccgaaaaagg gctccaagaa ggcggtgact
4501 aaggcgcaga agaaaggcgg caagaagcgc aagcgcagcc gcaaggagag ctattccatc
4561 tatgtgtaca aggttctgaa gcaggtccac cctgacaccg gcatttcgtc caaggccatg
4621 ggcatcatga attcgtttgt gaacgacatt ttcgagcgca tcgcaggtga ggcttcccgc
4681 ctggcgcatt acaacaagcg ctcgaccatc acctccaggg agatccagac ggccgtgcgc
4741 ctgctgctgc ctggggagtt ggccaagcac gccgtgtccg agggtactaa ggccatcacc
4801 aagtacacca gcgctaagga tccaccggtc gccaccatgg cctcctccga ggacgtcatc
4861 aaggagttca tgcgcttcaa ggtgcgcatg gagggctccg tgaacggcca cgagttcgag
4921 atcgagggcg agggcgaggg ccgcccctac gagggcaccc agaccgccaa gctgaaggtg
4981 accaagggcg gccccctgcc cttcgcctgg gacatcctgt cccctcagtt ccagtacggc
5041 tccaaggcct acgtgaagca ccccgccgac atccccgact acttgaagct gtccttcccc
5101 gagggcttca agtgggagcg cgtgatgaac ttcgaggacg gcggcgtggt gaccgtgacc
5161 caggactcct ccctgcagga cggcgagttc atctacaagg tgaagctgcg cggcaccaac
5221 ttcccctccg acggccccgt aatgcagaag aagaccatgg gctgggaggc ctccaccgag
5281 cggatgtacc ccgaggacgg cgccctgaag ggcgagatca agatgaggct gaagctgaag
5341 gacggcggcc actacgacgc cgaggtcaag accacctaca tggccaagaa gcccgtgcag
5401 ctgcccggcg cctacaagac cgacatcaag ctggacatca cctcccacaa cgaggactac
5461 accatcgtgg aacagtacga gcgcgccgag ggccgccact ccaccggcgc ctaagcggcc
5521 gcatcgatac cgtcgacctc gatcgagacc tagaaaaaca tggagcaatc acaagtagca
5581 atacagcagc taccaatgct gattgtgcct ggctagaagc acaagaggag gaggaggtgg
5641 gttttccagt cacacctcag gtacctttaa gaccaatgac ttacaaggca gctgtagatc
5701 ttagccactt tttaaaagaa aaggggggac tggaagggct aattcactcc caacgaagac
5761 aagatatcct tgatctgtgg atctaccaca cacaaggcta cttccctgat tggcagaact
5821 acacaccagg gccagggatc agatatccac tgacctttgg atggtgctac aagctagtac
5881 cagttgagca agagaaggta gaagaagcca atgaaggaga gaacacccgc ttgttacacc
5941 ctgtgagcct gcatgggatg gatgacccgg agagagaagt attagagtgg aggtttgaca
6001 gccgcctagc atttcatcac atggcccgag agctgcatcc ggactgtact gggtctctct
6061 ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc
6121 ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg
6181 gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc agcatgtgag
6241 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
6301 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
6361 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
6421 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
6481 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
6541 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
6601 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
6661 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
6721 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
6781 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
6841 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
6901 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
6961 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
7021 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
7081 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
7141 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
7201 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
7261 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
7321 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
7381 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
7441 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
7501 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
7561 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
7621 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
7681 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
7741 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
7801 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
7861 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
7921 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
7981 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
8041 ctgac
//
质粒菌株产品操作说明书
一、扩增流程
收到产品后，请先根据产品管壁标签来判断产品形式，并在扩增前准确查找该质粒菌株的抗性、感受态和培养温度。
1、质粒干粉（常温运输，存于-20度，90天保质期，请务必转化挑单克隆培养，不要直接使用和测序）
①收到质粒干粉后请先5000rpm离心1min，再加入20μl ddH2O去离子水溶解质粒；
②取1支100μl 感受态于冰上解冻10min，加入2μl质粒，再冰浴30min后，42℃热激60s，不要搅动，再冰浴2min；（从第二步开始均要在超净工作台中无菌操作）
③加入900μl无抗的LB液体培养基，180rpm震荡37℃培养45min；
④6000rpm离心5min，仅留100μl上清液重悬细菌沉淀，并涂布至目标质粒抗性的LB平板上；（可使用本平台的平板涂布专用玻璃珠进行涂布，可以比传统涂布方法获得更多转化子）
⑤将平板正向培养1h，再倒置37℃培养14h。如果要求是30度则培养20h；
（菌落过多则将质粒稀释后再转化。没有菌落则加入10μl质粒转化。另不要直接转表达感受态，要先转克隆感受态，重提质粒后再导入表达感受态）
⑥挑取单菌落至LB液体培养基中，加入对应抗生素，220rpm震荡培养14h，根据实验需要和质粒提取试剂盒说明书提取质粒。
2、甘油菌种（冰袋运输，存于-80℃，保质期90天，请务必划线挑单克隆培养）
四区划线后挑单菌落培养，酵母菌需要先液体复苏再四区划线，再挑单菌落液体培养。
3、穿刺菌种（冰袋运输，存于4℃，保质期7天）
穿刺接种，液体培养后四区划线，再挑单菌落液体培养。
4、菌落平板（冰袋运输，存于4℃，保质期7天）
直接挑取单菌落至液体培养基中。
5、液体质粒（冰袋运输，存于-20℃，保质期90天）
单独提取的液体质粒收到后可直接使用。
6、滤纸质粒（常温运输，存于-20度，90天保质期，请务必转化挑单克隆培养，不要直接使用和测序）
收到货后将滤纸画圈部分剪下放入EP管中，加100ul无菌水将滤纸浸湿并浸泡5min，吸取5ul质粒转化，离心全涂。
二、转化图片
| P2258/pSV40-Eif5a-m小鼠基因质粒 |
| P2259/pSV40-Tdrp-m小鼠基因质粒 |
| P2260/pSV40-Itm2b-m小鼠基因质粒 |
| P2261/pSV40-Mrpl38-m小鼠基因质粒 |
| P2262/pSV40-Rab6a-m小鼠基因质粒 |
| P2263/pSV40-Mdh1-m小鼠基因质粒 |
| P2264/pSV40-Sccpdh-m小鼠基因质粒 |
| P2265/pSV40-Haghl-m小鼠基因质粒 |
| P2266/pSV40-Fam19a1-m小鼠基因质粒 |
| P2267/pSV40-Fkbp4-m小鼠基因质粒 |
| P2268/pSV40-Gnas-m小鼠基因质粒 |
风险提示：丁香通仅作为第三方平台，为商家信息发布提供平台空间。用户咨询产品时请注意保护个人信息及财产安全，合理判断，谨慎选购商品，商家和用户对交易行为负责。对于医疗器械类产品，请先查证核实企业经营资质和医疗器械产品注册证情况。
pHIV-H2BmRFP慢病毒表达质粒
￥100 - 1000
