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- 详细信息
- 询价记录
- 技术资料
- 保存条件：
-20
- 保质期：
2年
- 英文名：
Plasmid#12251;pMDLg/pRRE
- 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：pMDLg-pRRE慢病毒包装质粒
别称: Plasmid#12251;pMDLg/pRRE
质粒属性
质粒简介
This 3rd generation LV packaging plasmid includes gag, coding for the virion main structural proteins; pol, responsible for the retrovirus-specific enzymes; and RRE, a binding site for the Rev protein which facilitates export of the RNA from the nucleus. This plasmid does not include Rev. The third generation packaging system offers maximal biosafety but is more cumbersome, as it involves the transfection of four different plasmids in the producer cells.
质粒图谱
质粒序列
LOCUS Exported 8890 bp ds-DNA circular SYN 01-SEP-2016
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS Untitled 5
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8890)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Thursday, September 1, 2016 from SnapGene Viewer 3.1.4
http://www.miaolingbio.com
FEATURES Location/Qualifiers
source 1..8890
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 22..401
/note="CMV enhancer"
/note="human cytomegalovirus immediate early enhancer"
promoter 402..605
/note="CMV promoter"
/note="human cytomegalovirus (CMV) immediate early
promoter"
intron 739..1214
/note="beta-globin intron"
/note="internally truncated intron from human beta-globin"
CDS 1298..2800
/codon_start=1
/gene="gag"
/product="gag protein from human immunodeficiency virus 1"
/note="HIV-1 gag"
/translation="MGARASVLSGGELDRWEKIRLRPGGKKKYKLKHIVWASRELERFA
VNPGLLETSEGCRQILGQLQPSLQTGSEELRSLYNTVATLYCVHQRIEIKDTKEALDKI
EEEQNKSKKKAQQAAADTGHSNQVSQNYPIVQNIQGQMVHQAISPRTLNAWVKVVEEKA
FSPEVIPMFSALSEGATPQDLNTMLNTVGGHQAAMQMLKETINEEAAEWDRVHPVHAGP
IAPGQMREPRGSDIAGTTSTLQEQIGWMTHNPPIPVGEIYKRWIILGLNKIVRMYSPTS
ILDIRQGPKEPFRDYVDRFYKTLRAEQASQEVKNWMTETLLVQNANPDCKTILKALGPG
ATLEEMMTACQGVGGPGHKARVLAEAMSQVTNPATIMIQKGNFRNQRKTVKCFNCGKEG
HIAKNCRAPRKKGCWKCGKEGHQMKDCTERQANFLGKIWPSHKGRPGNFLQSRPEPTAP
PEESFRFGEETTTPSQKQEPIDKELYPLASLRSLFGSDPSSQ"
CDS 2593..5604
/codon_start=1
/gene="pol"
/product="pol protein from human immunodeficiency virus 1"
/note="HIV-1 pol"
/translation="FFREDLAFPQGKAREFSSEQTRANSPTRRELQVWGRDNNSLSEAG
ADRQGTVSFSFPQITLWQRPLVTIKIGGQLKEALLDTGADDTVLEEMNLPGRWKPKMIG
GIGGFIKVRQYDQILIEICGHKAIGTVLVGPTPVNIIGRNLLTQIGCTLNFPISPIETV
PVKLKPGMDGPKVKQWPLTEEKIKALVEICTEMEKEGKISKIGPENPYNTPVFAIKKKD
STKWRKLVDFRELNKRTQDFWEVQLGIP-HPAGLKQKKSVTVLDVGDAYFSVPLDKDFRK
YTAFTIPSINNETPGIRYQYNVLPQGWKGSPAIFQCSMTKILEPFRKQNPDIVIYQYMD
DLYVGSDLEIGQHRTKIEELRQHLLRWGFTTPDKKHQKEPPFLWMGYELHPDKWTVQPI
VLPEKDSWTVNDIQKLVGKLNWASQIYAGIKVRQLCKLLRGTKALTEVVPLTEEAELEL
AENREILKEPVHGVYYDPSKDLIAEIQKQGQGQWTYQIYQEPFKNLKTGKYARMKGAHT
NDVKQLTEAVQKIATESIVIWGKTPKFKLPIQKETWEAWWTEYWQATWIPEWEFVNTPP
LVKLWYQLEKEPIIGAETFYVDGAANRETKLGKAGYVTDRGRQKVVPLTDTTNQKTELQ
AIHLALQDSGLEVNIVTDSQYALGIIQAQPDKSESELVSQIIEQLIKKEKVYLAWVPAH
KGIGGNEQVDKLVSAGIRKVLFLDGIDKAQEEHEKYHSNWRAMASDFNLPPVVAKEIVA
SCDKCQLKGEAMHGQVDCSPGIWQLDCTHLEGKVILVAVHVASGYIEAEVIPAETGQET
AYFLLKLAGRWPVKTVHTDNGSNFTSTTVKAACWWAGIKQEFGIPYNPQSQGVIESMNK
ELKKIIGQVRDQAEHLKTAVQMAVFIHNFKRKGGIGGYSAGERIVDIIATDIQTKELQK
QITKIQNFRVYYRDSRDPVWKGPAKLLWKGEGAVVIQDNSDIKVVPRRKAKIIRDYGKQ
MAGDDCVASRQDED"
misc_feature 5289..5406
/note="cPPT/CTS"
/note="central polypurine tract and central termination
sequence of HIV-1"
misc_feature 5629..5862
/note="RRE"
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
polyA_site 6093..6487
/note="beta-globin poly(A)"
/note="human beta-globin polyadenylation signal"
rep_origin complement(6997..7585)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(7756..8616)
/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(8617..8721)
/gene="bla"
/note="AmpR promoter"
ORIGIN
1 gtccaacatt accgccatgt tgacattgat tattgactag ttattaatag taatcaatta
61 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
121 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
181 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
241 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
301 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
361 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
421 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
481 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
541 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
601 gagctcgttt agtgaaccgt cagatcgcct ggagacgcca tccacgctgt tttgacctcc
661 atagaagaca ccgggaccga tccagcctcc cctcgaagct tacatgtggt accgagctcg
721 gatcctgaga acttcagggt gagtctatgg gacccttgat gttttctttc cccttctttt
781 ctatggttaa gttcatgtca taggaagggg agaagtaaca gggtacacat attgaccaaa
841 tcagggtaat tttgcatttg taattttaaa aaatgctttc ttcttttaat atactttttt
901 gtttatctta tttctaatac tttccctaat ctctttcttt cagggcaata atgatacaat
961 gtatcatgcc tctttgcacc attctaaaga ataacagtga taatttctgg gttaaggcaa
1021 tagcaatatt tctgcatata aatatttctg catataaatt gtaactgatg taagaggttt
1081 catattgcta atagcagcta caatccagct accattctgc ttttatttta tggttgggat
1141 aaggctggat tattctgagt ccaagctagg cccttttgct aatcatgttc atacctctta
1201 tcttcctccc acagctcctg ggcaacgtgc tggtctgtgt gctggcccat cactttggca
1261 aagcacgtga gatctgaatt cgagatctgc cgccgccatg ggtgcgagag cgtcagtatt
1321 aagcggggga gaattagatc gatgggaaaa aattcggtta aggccagggg gaaagaaaaa
1381 atataaatta aaacatatag tatgggcaag cagggagcta gaacgattcg cagttaatcc
1441 tggcctgtta gaaacatcag aaggctgtag acaaatactg ggacagctac aaccatccct
1501 tcagacagga tcagaagaac ttagatcatt atataataca gtagcaaccc tctattgtgt
1561 gcatcaaagg atagagataa aagacaccaa ggaagcttta gacaagatag aggaagagca
1621 aaacaaaagt aagaaaaaag cacagcaagc agcagctgac acaggacaca gcaatcaggt
1681 cagccaaaat taccctatag tgcagaacat ccaggggcaa atggtacatc aggccatatc
1741 acctagaact ttaaatgcat gggtaaaagt agtagaagag aaggctttca gcccagaagt
1801 gatacccatg ttttcagcat tatcagaagg agccacccca caagatttaa acaccatgct
1861 aaacacagtg gggggacatc aagcagccat gcaaatgtta aaagagacca tcaatgagga
1921 agctgcagaa tgggatagag tgcatccagt gcatgcaggg cctattgcac caggccagat
1981 gagagaacca aggggaagtg acatagcagg aactactagt acccttcagg aacaaatagg
2041 atggatgaca cataatccac ctatcccagt aggagaaatc tataaaagat ggataatcct
2101 gggattaaat aaaatagtaa gaatgtatag ccctaccagc attctggaca taagacaagg
2161 accaaaggaa ccctttagag actatgtaga ccgattctat aaaactctaa gagccgagca
2221 agcttcacaa gaggtaaaaa attggatgac agaaaccttg ttggtccaaa atgcgaaccc
2281 agattgtaag actattttaa aagcattggg accaggagcg acactagaag aaatgatgac
2341 agcatgtcag ggagtggggg gacccggcca taaagcaaga gttttggctg aagcaatgag
2401 ccaagtaaca aatccagcta ccataatgat acagaaaggc aattttagga accaaagaaa
2461 gactgttaag tgtttcaatt gtggcaaaga agggcacata gccaaaaatt gcagggcccc
2521 taggaaaaag ggctgttgga aatgtggaaa ggaaggacac caaatgaaag attgtactga
2581 gagacaggct aattttttag ggaagatctg gccttcccac aagggaaggc cagggaattt
2641 tcttcagagc agaccagagc caacagcccc accagaagag agcttcaggt ttggggaaga
2701 gacaacaact ccctctcaga agcaggagcc gatagacaag gaactgtatc ctttagcttc
2761 cctcagatca ctctttggca gcgacccctc gtcacaataa agataggggg gcaattaaag
2821 gaagctctat tagatacagg agcagatgat acagtattag aagaaatgaa tttgccagga
2881 agatggaaac caaaaatgat agggggaatt ggaggtttta tcaaagtaag acagtatgat
2941 cagatactca tagaaatctg cggacataaa gctataggta cagtattagt aggacctaca
3001 cctgtcaaca taattggaag aaatctgttg actcagattg gctgcacttt aaattttccc
3061 attagtccta ttgagactgt accagtaaaa ttaaagccag gaatggatgg cccaaaagtt
3121 aaacaatggc cattgacaga agaaaaaata aaagcattag tagaaatttg tacagaaatg
3181 gaaaaggaag gaaaaatttc aaaaattggg cctgaaaatc catacaatac tccagtattt
3241 gccataaaga aaaaagacag tactaaatgg agaaaattag tagatttcag agaacttaat
3301 aagagaactc aagatttctg ggaagttcaa ttaggaatac cacatcctgc agggttaaaa
3361 cagaaaaaat cagtaacagt actggatgtg ggcgatgcat atttttcagt tcccttagat
3421 aaagacttca ggaagtatac tgcatttacc atacctagta taaacaatga gacaccaggg
3481 attagatatc agtacaatgt gcttccacag ggatggaaag gatcaccagc aatattccag
3541 tgtagcatga caaaaatctt agagcctttt agaaaacaaa atccagacat agtcatctat
3601 caatacatgg atgatttgta tgtaggatct gacttagaaa tagggcagca tagaacaaaa
3661 atagaggaac tgagacaaca tctgttgagg tggggattta ccacaccaga caaaaaacat
3721 cagaaagaac ctccattcct ttggatgggt tatgaactcc atcctgataa atggacagta
3781 cagcctatag tgctgccaga aaaggacagc tggactgtca atgacataca gaaattagtg
3841 ggaaaattga attgggcaag tcagatttat gcagggatta aagtaaggca attatgtaaa
3901 cttcttaggg gaaccaaagc actaacagaa gtagtaccac taacagaaga agcagagcta
3961 gaactggcag aaaacaggga gattctaaaa gaaccggtac atggagtgta ttatgaccca
4021 tcaaaagact taatagcaga aatacagaag caggggcaag gccaatggac atatcaaatt
4081 tatcaagagc catttaaaaa tctgaaaaca ggaaagtatg caagaatgaa gggtgcccac
4141 actaatgatg tgaaacaatt aacagaggca gtacaaaaaa tagccacaga aagcatagta
4201 atatggggaa agactcctaa atttaaatta cccatacaaa aggaaacatg ggaagcatgg
4261 tggacagagt attggcaagc cacctggatt cctgagtggg agtttgtcaa tacccctccc
4321 ttagtgaagt tatggtacca gttagagaaa gaacccataa taggagcaga aactttctat
4381 gtagatgggg cagccaatag ggaaactaaa ttaggaaaag caggatatgt aactgacaga
4441 ggaagacaaa aagttgtccc cctaacggac acaacaaatc agaagactga gttacaagca
4501 attcatctag ctttgcagga ttcgggatta gaagtaaaca tagtgacaga ctcacaatat
4561 gcattgggaa tcattcaagc acaaccagat aagagtgaat cagagttagt cagtcaaata
4621 atagagcagt taataaaaaa ggaaaaagtc tacctggcat gggtaccagc acacaaagga
4681 attggaggaa atgaacaagt agataaattg gtcagtgctg gaatcaggaa agtactattt
4741 ttagatggaa tagataaggc ccaagaagaa catgagaaat atcacagtaa ttggagagca
4801 atggctagtg attttaacct accacctgta gtagcaaaag aaatagtagc cagctgtgat
4861 aaatgtcagc taaaagggga agccatgcat ggacaagtag actgtagccc aggaatatgg
4921 cagctagatt gtacacattt agaaggaaaa gttatcttgg tagcagttca tgtagccagt
4981 ggatatatag aagcagaagt aattccagca gagacagggc aagaaacagc atacttcctc
5041 ttaaaattag caggaagatg gccagtaaaa acagtacata cagacaatgg cagcaatttc
5101 accagtacta cagttaaggc cgcctgttgg tgggcgggga tcaagcagga atttggcatt
5161 ccctacaatc cccaaagtca aggagtaata gaatctatga ataaagaatt aaagaaaatt
5221 ataggacagg taagagatca ggctgaacat cttaagacag cagtacaaat ggcagtattc
5281 atccacaatt ttaaaagaaa aggggggatt ggggggtaca gtgcagggga aagaatagta
5341 gacataatag caacagacat acaaactaaa gaattacaaa aacaaattac aaaaattcaa
5401 aattttcggg tttattacag ggacagcaga gatccagttt ggaaaggacc agcaaagctc
5461 ctctggaaag gtgaaggggc agtagtaata caagataata gtgacataaa agtagtgcca
5521 agaagaaaag caaagatcat cagggattat ggaaaacaga tggcaggtga tgattgtgtg
5581 gcaagtagac aggatgagga ttaacacatg gaattccgga gcggccgcag gagctttgtt
5641 ccttgggttc ttgggagcag caggaagcac tatgggcgca gcgtcaatga cgctgacggt
5701 acaggccaga caattattgt ctggtatagt gcagcagcag aacaatttgc tgagggctat
5761 tgaggcgcaa cagcatctgt tgcaactcac agtctggggc atcaagcagc tccaggcaag
5821 aatcctggct gtggaaagat acctaaagga tcaacagctc ctggggattt ggggttgctc
5881 tggaaaactc atttgcacca ctgctgtgcc ttggaatgct agttggagta ataaatctct
5941 ggaacagatt tggaatcaca cgacctggat ggagtgggac agagaaatta acaattacac
6001 aagcttccgc ggaattcacc ccaccagtgc aggctgccta tcagaaagtg gtggctggtg
6061 tggctaatgc cctggcccac aagtatcact aagctcgctt tcttgctgtc caatttctat
6121 taaaggttcc tttgttccct aagtccaact actaaactgg gggatattat gaagggcctt
6181 gagcatctgg attctgccta ataaaaaaca tttattttca ttgcaatgat gtatttaaat
6241 tatttctgaa tattttacta aaaagggaat gtgggaggtc agtgcattta aaacataaag
6301 aaatgaagag ctagttcaaa ccttgggaaa atacactata tcttaaactc catgaaagaa
6361 ggtgaggctg caaacagcta atgcacattg gcaacagccc ctgatgccta tgccttattc
6421 atccctcaga aaaggattca agtagaggct tgatttggag gttaaagttt tgctatgctg
6481 tattttacat tacttattgt tttagctgtc ctcatgaatg tcttttcact acccatttgc
6541 ttatcctgca tctctcagcc ttgactccac tcagttctct tgcttagaga taccaccttt
6601 cccctgaagt gttccttcca tgttttacgg cgagatggtt tctcctcgcc tggccactca
6661 gccttagttg tctctgttgt cttatagagg tctacttgaa gaaggaaaaa cagggggcat
6721 ggtttgactg tcctgtgagc ccttcttccc tgcctccccc actcacagtg acccggaatc
6781 cctcgacatg gcagtctagc actagtgcgg ccgcagatct gcttcctcgc tcactgactc
6841 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
6901 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
6961 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
7021 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
7081 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
7141 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
7201 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
7261 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
7321 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
7381 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
7441 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
7501 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
7561 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
7621 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
7681 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
7741 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
7801 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
7861 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
7921 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
7981 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
8041 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
8101 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
8161 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
8221 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
8281 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
8341 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
8401 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
8461 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
8521 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
8581 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
8641 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
8701 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tggatcccct
8761 gagggggccc ccatgggcta gaggatccgg cctcggcctc tgcataaata aaaaaaatta
8821 gtcagccatg agcttggccc attgcatacg ttgtatccat atcataatat gtacatttat
8881 attggctcat
//
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：pMDLg-pRRE慢病毒包装质粒
别称: Plasmid#12251;pMDLg/pRRE
| 启动子: | CMV |
|---|---|
| 复制子: | pUC |
| 终止子: | β-globin poly(A) |
| 质粒大小: | 8890bp |
| 原核抗性: | Amp |
| 克隆菌株: | DH5a |
| 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： | 包装辅助 |
| 基因种属： | 病毒 |
| 基因类型： | ORF |
| 原核抗性： | Amp |
| 筛选标记： | |
| 荧光蛋白： |
质粒简介
This 3rd generation LV packaging plasmid includes gag, coding for the virion main structural proteins; pol, responsible for the retrovirus-specific enzymes; and RRE, a binding site for the Rev protein which facilitates export of the RNA from the nucleus. This plasmid does not include Rev. The third generation packaging system offers maximal biosafety but is more cumbersome, as it involves the transfection of four different plasmids in the producer cells.
质粒图谱
质粒序列
LOCUS Exported 8890 bp ds-DNA circular SYN 01-SEP-2016
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS Untitled 5
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8890)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Thursday, September 1, 2016 from SnapGene Viewer 3.1.4
http://www.miaolingbio.com
FEATURES Location/Qualifiers
source 1..8890
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 22..401
/note="CMV enhancer"
/note="human cytomegalovirus immediate early enhancer"
promoter 402..605
/note="CMV promoter"
/note="human cytomegalovirus (CMV) immediate early
promoter"
intron 739..1214
/note="beta-globin intron"
/note="internally truncated intron from human beta-globin"
CDS 1298..2800
/codon_start=1
/gene="gag"
/product="gag protein from human immunodeficiency virus 1"
/note="HIV-1 gag"
/translation="MGARASVLSGGELDRWEKIRLRPGGKKKYKLKHIVWASRELERFA
VNPGLLETSEGCRQILGQLQPSLQTGSEELRSLYNTVATLYCVHQRIEIKDTKEALDKI
EEEQNKSKKKAQQAAADTGHSNQVSQNYPIVQNIQGQMVHQAISPRTLNAWVKVVEEKA
FSPEVIPMFSALSEGATPQDLNTMLNTVGGHQAAMQMLKETINEEAAEWDRVHPVHAGP
IAPGQMREPRGSDIAGTTSTLQEQIGWMTHNPPIPVGEIYKRWIILGLNKIVRMYSPTS
ILDIRQGPKEPFRDYVDRFYKTLRAEQASQEVKNWMTETLLVQNANPDCKTILKALGPG
ATLEEMMTACQGVGGPGHKARVLAEAMSQVTNPATIMIQKGNFRNQRKTVKCFNCGKEG
HIAKNCRAPRKKGCWKCGKEGHQMKDCTERQANFLGKIWPSHKGRPGNFLQSRPEPTAP
PEESFRFGEETTTPSQKQEPIDKELYPLASLRSLFGSDPSSQ"
CDS 2593..5604
/codon_start=1
/gene="pol"
/product="pol protein from human immunodeficiency virus 1"
/note="HIV-1 pol"
/translation="FFREDLAFPQGKAREFSSEQTRANSPTRRELQVWGRDNNSLSEAG
ADRQGTVSFSFPQITLWQRPLVTIKIGGQLKEALLDTGADDTVLEEMNLPGRWKPKMIG
GIGGFIKVRQYDQILIEICGHKAIGTVLVGPTPVNIIGRNLLTQIGCTLNFPISPIETV
PVKLKPGMDGPKVKQWPLTEEKIKALVEICTEMEKEGKISKIGPENPYNTPVFAIKKKD
STKWRKLVDFRELNKRTQDFWEVQLGIP-HPAGLKQKKSVTVLDVGDAYFSVPLDKDFRK
YTAFTIPSINNETPGIRYQYNVLPQGWKGSPAIFQCSMTKILEPFRKQNPDIVIYQYMD
DLYVGSDLEIGQHRTKIEELRQHLLRWGFTTPDKKHQKEPPFLWMGYELHPDKWTVQPI
VLPEKDSWTVNDIQKLVGKLNWASQIYAGIKVRQLCKLLRGTKALTEVVPLTEEAELEL
AENREILKEPVHGVYYDPSKDLIAEIQKQGQGQWTYQIYQEPFKNLKTGKYARMKGAHT
NDVKQLTEAVQKIATESIVIWGKTPKFKLPIQKETWEAWWTEYWQATWIPEWEFVNTPP
LVKLWYQLEKEPIIGAETFYVDGAANRETKLGKAGYVTDRGRQKVVPLTDTTNQKTELQ
AIHLALQDSGLEVNIVTDSQYALGIIQAQPDKSESELVSQIIEQLIKKEKVYLAWVPAH
KGIGGNEQVDKLVSAGIRKVLFLDGIDKAQEEHEKYHSNWRAMASDFNLPPVVAKEIVA
SCDKCQLKGEAMHGQVDCSPGIWQLDCTHLEGKVILVAVHVASGYIEAEVIPAETGQET
AYFLLKLAGRWPVKTVHTDNGSNFTSTTVKAACWWAGIKQEFGIPYNPQSQGVIESMNK
ELKKIIGQVRDQAEHLKTAVQMAVFIHNFKRKGGIGGYSAGERIVDIIATDIQTKELQK
QITKIQNFRVYYRDSRDPVWKGPAKLLWKGEGAVVIQDNSDIKVVPRRKAKIIRDYGKQ
MAGDDCVASRQDED"
misc_feature 5289..5406
/note="cPPT/CTS"
/note="central polypurine tract and central termination
sequence of HIV-1"
misc_feature 5629..5862
/note="RRE"
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
polyA_site 6093..6487
/note="beta-globin poly(A)"
/note="human beta-globin polyadenylation signal"
rep_origin complement(6997..7585)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(7756..8616)
/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(8617..8721)
/gene="bla"
/note="AmpR promoter"
ORIGIN
1 gtccaacatt accgccatgt tgacattgat tattgactag ttattaatag taatcaatta
61 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
121 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
181 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
241 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
301 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
361 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
421 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
481 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
541 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
601 gagctcgttt agtgaaccgt cagatcgcct ggagacgcca tccacgctgt tttgacctcc
661 atagaagaca ccgggaccga tccagcctcc cctcgaagct tacatgtggt accgagctcg
721 gatcctgaga acttcagggt gagtctatgg gacccttgat gttttctttc cccttctttt
781 ctatggttaa gttcatgtca taggaagggg agaagtaaca gggtacacat attgaccaaa
841 tcagggtaat tttgcatttg taattttaaa aaatgctttc ttcttttaat atactttttt
901 gtttatctta tttctaatac tttccctaat ctctttcttt cagggcaata atgatacaat
961 gtatcatgcc tctttgcacc attctaaaga ataacagtga taatttctgg gttaaggcaa
1021 tagcaatatt tctgcatata aatatttctg catataaatt gtaactgatg taagaggttt
1081 catattgcta atagcagcta caatccagct accattctgc ttttatttta tggttgggat
1141 aaggctggat tattctgagt ccaagctagg cccttttgct aatcatgttc atacctctta
1201 tcttcctccc acagctcctg ggcaacgtgc tggtctgtgt gctggcccat cactttggca
1261 aagcacgtga gatctgaatt cgagatctgc cgccgccatg ggtgcgagag cgtcagtatt
1321 aagcggggga gaattagatc gatgggaaaa aattcggtta aggccagggg gaaagaaaaa
1381 atataaatta aaacatatag tatgggcaag cagggagcta gaacgattcg cagttaatcc
1441 tggcctgtta gaaacatcag aaggctgtag acaaatactg ggacagctac aaccatccct
1501 tcagacagga tcagaagaac ttagatcatt atataataca gtagcaaccc tctattgtgt
1561 gcatcaaagg atagagataa aagacaccaa ggaagcttta gacaagatag aggaagagca
1621 aaacaaaagt aagaaaaaag cacagcaagc agcagctgac acaggacaca gcaatcaggt
1681 cagccaaaat taccctatag tgcagaacat ccaggggcaa atggtacatc aggccatatc
1741 acctagaact ttaaatgcat gggtaaaagt agtagaagag aaggctttca gcccagaagt
1801 gatacccatg ttttcagcat tatcagaagg agccacccca caagatttaa acaccatgct
1861 aaacacagtg gggggacatc aagcagccat gcaaatgtta aaagagacca tcaatgagga
1921 agctgcagaa tgggatagag tgcatccagt gcatgcaggg cctattgcac caggccagat
1981 gagagaacca aggggaagtg acatagcagg aactactagt acccttcagg aacaaatagg
2041 atggatgaca cataatccac ctatcccagt aggagaaatc tataaaagat ggataatcct
2101 gggattaaat aaaatagtaa gaatgtatag ccctaccagc attctggaca taagacaagg
2161 accaaaggaa ccctttagag actatgtaga ccgattctat aaaactctaa gagccgagca
2221 agcttcacaa gaggtaaaaa attggatgac agaaaccttg ttggtccaaa atgcgaaccc
2281 agattgtaag actattttaa aagcattggg accaggagcg acactagaag aaatgatgac
2341 agcatgtcag ggagtggggg gacccggcca taaagcaaga gttttggctg aagcaatgag
2401 ccaagtaaca aatccagcta ccataatgat acagaaaggc aattttagga accaaagaaa
2461 gactgttaag tgtttcaatt gtggcaaaga agggcacata gccaaaaatt gcagggcccc
2521 taggaaaaag ggctgttgga aatgtggaaa ggaaggacac caaatgaaag attgtactga
2581 gagacaggct aattttttag ggaagatctg gccttcccac aagggaaggc cagggaattt
2641 tcttcagagc agaccagagc caacagcccc accagaagag agcttcaggt ttggggaaga
2701 gacaacaact ccctctcaga agcaggagcc gatagacaag gaactgtatc ctttagcttc
2761 cctcagatca ctctttggca gcgacccctc gtcacaataa agataggggg gcaattaaag
2821 gaagctctat tagatacagg agcagatgat acagtattag aagaaatgaa tttgccagga
2881 agatggaaac caaaaatgat agggggaatt ggaggtttta tcaaagtaag acagtatgat
2941 cagatactca tagaaatctg cggacataaa gctataggta cagtattagt aggacctaca
3001 cctgtcaaca taattggaag aaatctgttg actcagattg gctgcacttt aaattttccc
3061 attagtccta ttgagactgt accagtaaaa ttaaagccag gaatggatgg cccaaaagtt
3121 aaacaatggc cattgacaga agaaaaaata aaagcattag tagaaatttg tacagaaatg
3181 gaaaaggaag gaaaaatttc aaaaattggg cctgaaaatc catacaatac tccagtattt
3241 gccataaaga aaaaagacag tactaaatgg agaaaattag tagatttcag agaacttaat
3301 aagagaactc aagatttctg ggaagttcaa ttaggaatac cacatcctgc agggttaaaa
3361 cagaaaaaat cagtaacagt actggatgtg ggcgatgcat atttttcagt tcccttagat
3421 aaagacttca ggaagtatac tgcatttacc atacctagta taaacaatga gacaccaggg
3481 attagatatc agtacaatgt gcttccacag ggatggaaag gatcaccagc aatattccag
3541 tgtagcatga caaaaatctt agagcctttt agaaaacaaa atccagacat agtcatctat
3601 caatacatgg atgatttgta tgtaggatct gacttagaaa tagggcagca tagaacaaaa
3661 atagaggaac tgagacaaca tctgttgagg tggggattta ccacaccaga caaaaaacat
3721 cagaaagaac ctccattcct ttggatgggt tatgaactcc atcctgataa atggacagta
3781 cagcctatag tgctgccaga aaaggacagc tggactgtca atgacataca gaaattagtg
3841 ggaaaattga attgggcaag tcagatttat gcagggatta aagtaaggca attatgtaaa
3901 cttcttaggg gaaccaaagc actaacagaa gtagtaccac taacagaaga agcagagcta
3961 gaactggcag aaaacaggga gattctaaaa gaaccggtac atggagtgta ttatgaccca
4021 tcaaaagact taatagcaga aatacagaag caggggcaag gccaatggac atatcaaatt
4081 tatcaagagc catttaaaaa tctgaaaaca ggaaagtatg caagaatgaa gggtgcccac
4141 actaatgatg tgaaacaatt aacagaggca gtacaaaaaa tagccacaga aagcatagta
4201 atatggggaa agactcctaa atttaaatta cccatacaaa aggaaacatg ggaagcatgg
4261 tggacagagt attggcaagc cacctggatt cctgagtggg agtttgtcaa tacccctccc
4321 ttagtgaagt tatggtacca gttagagaaa gaacccataa taggagcaga aactttctat
4381 gtagatgggg cagccaatag ggaaactaaa ttaggaaaag caggatatgt aactgacaga
4441 ggaagacaaa aagttgtccc cctaacggac acaacaaatc agaagactga gttacaagca
4501 attcatctag ctttgcagga ttcgggatta gaagtaaaca tagtgacaga ctcacaatat
4561 gcattgggaa tcattcaagc acaaccagat aagagtgaat cagagttagt cagtcaaata
4621 atagagcagt taataaaaaa ggaaaaagtc tacctggcat gggtaccagc acacaaagga
4681 attggaggaa atgaacaagt agataaattg gtcagtgctg gaatcaggaa agtactattt
4741 ttagatggaa tagataaggc ccaagaagaa catgagaaat atcacagtaa ttggagagca
4801 atggctagtg attttaacct accacctgta gtagcaaaag aaatagtagc cagctgtgat
4861 aaatgtcagc taaaagggga agccatgcat ggacaagtag actgtagccc aggaatatgg
4921 cagctagatt gtacacattt agaaggaaaa gttatcttgg tagcagttca tgtagccagt
4981 ggatatatag aagcagaagt aattccagca gagacagggc aagaaacagc atacttcctc
5041 ttaaaattag caggaagatg gccagtaaaa acagtacata cagacaatgg cagcaatttc
5101 accagtacta cagttaaggc cgcctgttgg tgggcgggga tcaagcagga atttggcatt
5161 ccctacaatc cccaaagtca aggagtaata gaatctatga ataaagaatt aaagaaaatt
5221 ataggacagg taagagatca ggctgaacat cttaagacag cagtacaaat ggcagtattc
5281 atccacaatt ttaaaagaaa aggggggatt ggggggtaca gtgcagggga aagaatagta
5341 gacataatag caacagacat acaaactaaa gaattacaaa aacaaattac aaaaattcaa
5401 aattttcggg tttattacag ggacagcaga gatccagttt ggaaaggacc agcaaagctc
5461 ctctggaaag gtgaaggggc agtagtaata caagataata gtgacataaa agtagtgcca
5521 agaagaaaag caaagatcat cagggattat ggaaaacaga tggcaggtga tgattgtgtg
5581 gcaagtagac aggatgagga ttaacacatg gaattccgga gcggccgcag gagctttgtt
5641 ccttgggttc ttgggagcag caggaagcac tatgggcgca gcgtcaatga cgctgacggt
5701 acaggccaga caattattgt ctggtatagt gcagcagcag aacaatttgc tgagggctat
5761 tgaggcgcaa cagcatctgt tgcaactcac agtctggggc atcaagcagc tccaggcaag
5821 aatcctggct gtggaaagat acctaaagga tcaacagctc ctggggattt ggggttgctc
5881 tggaaaactc atttgcacca ctgctgtgcc ttggaatgct agttggagta ataaatctct
5941 ggaacagatt tggaatcaca cgacctggat ggagtgggac agagaaatta acaattacac
6001 aagcttccgc ggaattcacc ccaccagtgc aggctgccta tcagaaagtg gtggctggtg
6061 tggctaatgc cctggcccac aagtatcact aagctcgctt tcttgctgtc caatttctat
6121 taaaggttcc tttgttccct aagtccaact actaaactgg gggatattat gaagggcctt
6181 gagcatctgg attctgccta ataaaaaaca tttattttca ttgcaatgat gtatttaaat
6241 tatttctgaa tattttacta aaaagggaat gtgggaggtc agtgcattta aaacataaag
6301 aaatgaagag ctagttcaaa ccttgggaaa atacactata tcttaaactc catgaaagaa
6361 ggtgaggctg caaacagcta atgcacattg gcaacagccc ctgatgccta tgccttattc
6421 atccctcaga aaaggattca agtagaggct tgatttggag gttaaagttt tgctatgctg
6481 tattttacat tacttattgt tttagctgtc ctcatgaatg tcttttcact acccatttgc
6541 ttatcctgca tctctcagcc ttgactccac tcagttctct tgcttagaga taccaccttt
6601 cccctgaagt gttccttcca tgttttacgg cgagatggtt tctcctcgcc tggccactca
6661 gccttagttg tctctgttgt cttatagagg tctacttgaa gaaggaaaaa cagggggcat
6721 ggtttgactg tcctgtgagc ccttcttccc tgcctccccc actcacagtg acccggaatc
6781 cctcgacatg gcagtctagc actagtgcgg ccgcagatct gcttcctcgc tcactgactc
6841 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
6901 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
6961 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
7021 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
7081 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
7141 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
7201 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
7261 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
7321 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
7381 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
7441 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
7501 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
7561 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
7621 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
7681 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
7741 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
7801 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
7861 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
7921 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
7981 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
8041 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
8101 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
8161 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
8221 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
8281 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
8341 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
8401 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
8461 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
8521 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
8581 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
8641 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
8701 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tggatcccct
8761 gagggggccc ccatgggcta gaggatccgg cctcggcctc tgcataaata aaaaaaatta
8821 gtcagccatg agcttggccc attgcatacg ttgtatccat atcataatat gtacatttat
8881 attggctcat
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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
| P1941/pENTR223-CLEC4D人源基因质粒 |
| P1942/pENTR223-GNA11人源基因质粒 |
| P1943/pENTR223-FAD104人源基因质粒 |
| P1944/pENTR223-XAGE1A人源基因质粒 |
| P1945/pENTR223-PMS1-T477N人源基因质粒 |
| P1946/pENTR223-REG1A人源基因质粒 |
| P1947/pENTR223-IL20RB(427-936)人源基因质粒 |
| P1948/pENTR223-TMEM208人源基因质粒 |
| P1949/pENTR223-DPYD人源基因质粒 |
| P1950/pENTR223-ABHD4人源基因质粒 |
| P1951/pENTR223-SDHC-T491A-G499A人源基因质粒 |
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pMDLg-pRRE慢病毒包装质粒
￥100 - 1000
