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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：
pLP-1;pGag/Pol
- 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：pLP1慢病毒包装质粒
别称: pLP-1;pGag/Pol
质粒属性
质粒简介
PLP1 (Proteolipid Protein 1) is a Protein Coding gene. Diseases associated with PLP1 include pelizaeus-merzbacher disease and spastic paraplegia 2, x-linked. Among its related pathways are Glial Cell Differentiation and SIDS Susceptibility Pathways. GO annotations related to this gene include structural molecule activity and structural constituent of myelin sheath. An important paralog of this gene is GPM6B
质粒图谱
质粒序列
LOCUS Exported 8889 bp ds-DNA circular SYN 27-AUG-2016
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS Untitled 32
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8889)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Saturday, August 27, 2016 from SnapGene Viewer 3.1.4
http://www.miaolingbio.com
FEATURES Location/Qualifiers
source 1..8889
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 79..458
/note="CMV enhancer"
/note="human cytomegalovirus immediate early enhancer"
promoter 459..662
/note="CMV promoter"
/note="human cytomegalovirus (CMV) immediate early
promoter"
intron 796..1271
/note="beta-globin intron"
/note="internally truncated intron from human beta-globin"
CDS 1355..2857
/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 2650..5661
/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 5346..5463
/note="cPPT/CTS"
/note="central polypurine tract and central termination
sequence of HIV-1"
misc_feature 5686..5919
/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 6150..6544
/note="beta-globin poly(A)"
/note="human beta-globin polyadenylation signal"
rep_origin complement(7054..7642)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(7813..8673)
/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(8674..8778)
/gene="bla"
/note="AmpR promoter"
ORIGIN
1 ttggcccatt gcatacgttg tatccatatc ataatatgta catttatatt ggctcatgtc
61 caacattacc gccatgttga cattgattat tgactagtta ttaatagtaa tcaattacgg
121 ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc
181 cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca
241 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg
301 cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg
361 acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt
421 ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca
481 tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg
541 tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact
601 ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag
661 ctcgtttagt gaaccgtcag atcgcctgga gacgccatcc acgctgtttt gacctccata
721 gaagacaccg ggaccgatcc agcctcccct cgaagcttac atgtggtacc gagctcggat
781 cctgagaact tcagggtgag tctatgggac ccttgatgtt ttctttcccc ttcttttcta
841 tggttaagtt catgtcatag gaaggggaga agtaacaggg tacacatatt gaccaaatca
901 gggtaatttt gcatttgtaa ttttaaaaaa tgctttcttc ttttaatata cttttttgtt
961 tatcttattt ctaatacttt ccctaatctc tttctttcag ggcaataatg atacaatgta
1021 tcatgcctct ttgcaccatt ctaaagaata acagtgataa tttctgggtt aaggcaatag
1081 caatatttct gcatataaat atttctgcat ataaattgta actgatgtaa gaggtttcat
1141 attgctaata gcagctacaa tccagctacc attctgcttt tattttatgg ttgggataag
1201 gctggattat tctgagtcca agctaggccc ttttgctaat catgttcata cctcttatct
1261 tcctcccaca gctcctgggc aacgtgctgg tctgtgtgct ggcccatcac tttggcaaag
1321 cacgtgagat ctgaattcga gatctgccgc cgccatgggt gcgagagcgt cagtattaag
1381 cgggggagaa ttagatcgat gggaaaaaat tcggttaagg ccagggggaa agaaaaaata
1441 taaattaaaa catatagtat gggcaagcag ggagctagaa cgattcgcag ttaatcctgg
1501 cctgttagaa acatcagaag gctgtagaca aatactggga cagctacaac catcccttca
1561 gacaggatca gaagaactta gatcattata taatacagta gcaaccctct attgtgtgca
1621 tcaaaggata gagataaaag acaccaagga agctttagac aagatagagg aagagcaaaa
1681 caaaagtaag aaaaaagcac agcaagcagc agctgacaca ggacacagca atcaggtcag
1741 ccaaaattac cctatagtgc agaacatcca ggggcaaatg gtacatcagg ccatatcacc
1801 tagaacttta aatgcatggg taaaagtagt agaagagaag gctttcagcc cagaagtgat
1861 acccatgttt tcagcattat cagaaggagc caccccacaa gatttaaaca ccatgctaaa
1921 cacagtgggg ggacatcaag cagccatgca aatgttaaaa gagaccatca atgaggaagc
1981 tgcagaatgg gatagagtgc atccagtgca tgcagggcct attgcaccag gccagatgag
2041 agaaccaagg ggaagtgaca tagcaggaac tactagtacc cttcaggaac aaataggatg
2101 gatgacacat aatccaccta tcccagtagg agaaatctat aaaagatgga taatcctggg
2161 attaaataaa atagtaagaa tgtatagccc taccagcatt ctggacataa gacaaggacc
2221 aaaggaaccc tttagagact atgtagaccg attctataaa actctaagag ccgagcaagc
2281 ttcacaagag gtaaaaaatt ggatgacaga aaccttgttg gtccaaaatg cgaacccaga
2341 ttgtaagact attttaaaag cattgggacc aggagcgaca ctagaagaaa tgatgacagc
2401 atgtcaggga gtggggggac ccggccataa agcaagagtt ttggctgaag caatgagcca
2461 agtaacaaat ccagctacca taatgataca gaaaggcaat tttaggaacc aaagaaagac
2521 tgttaagtgt ttcaattgtg gcaaagaagg gcacatagcc aaaaattgca gggcccctag
2581 gaaaaagggc tgttggaaat gtggaaagga aggacaccaa atgaaagatt gtactgagag
2641 acaggctaat tttttaggga agatctggcc ttcccacaag ggaaggccag ggaattttct
2701 tcagagcaga ccagagccaa cagccccacc agaagagagc ttcaggtttg gggaagagac
2761 aacaactccc tctcagaagc aggagccgat agacaaggaa ctgtatcctt tagcttccct
2821 cagatcactc tttggcagcg acccctcgtc acaataaaga taggggggca attaaaggaa
2881 gctctattag atacaggagc agatgataca gtattagaag aaatgaattt gccaggaaga
2941 tggaaaccaa aaatgatagg gggaattgga ggttttatca aagtaagaca gtatgatcag
3001 atactcatag aaatctgcgg acataaagct ataggtacag tattagtagg acctacacct
3061 gtcaacataa ttggaagaaa tctgttgact cagattggct gcactttaaa ttttcccatt
3121 agtcctattg agactgtacc agtaaaatta aagccaggaa tggatggccc aaaagttaaa
3181 caatggccat tgacagaaga aaaaataaaa gcattagtag aaatttgtac agaaatggaa
3241 aaggaaggaa aaatttcaaa aattgggcct gaaaatccat acaatactcc agtatttgcc
3301 ataaagaaaa aagacagtac taaatggaga aaattagtag atttcagaga acttaataag
3361 agaactcaag atttctggga agttcaatta ggaataccac atcctgcagg gttaaaacag
3421 aaaaaatcag taacagtact ggatgtgggc gatgcatatt tttcagttcc cttagataaa
3481 gacttcagga agtatactgc atttaccata cctagtataa acaatgagac accagggatt
3541 agatatcagt acaatgtgct tccacaggga tggaaaggat caccagcaat attccagtgt
3601 agcatgacaa aaatcttaga gccttttaga aaacaaaatc cagacatagt catctatcaa
3661 tacatggatg atttgtatgt aggatctgac ttagaaatag ggcagcatag aacaaaaata
3721 gaggaactga gacaacatct gttgaggtgg ggatttacca caccagacaa aaaacatcag
3781 aaagaacctc cattcctttg gatgggttat gaactccatc ctgataaatg gacagtacag
3841 cctatagtgc tgccagaaaa ggacagctgg actgtcaatg acatacagaa attagtggga
3901 aaattgaatt gggcaagtca gatttatgca gggattaaag taaggcaatt atgtaaactt
3961 cttaggggaa ccaaagcact aacagaagta gtaccactaa cagaagaagc agagctagaa
4021 ctggcagaaa acagggagat tctaaaagaa ccggtacatg gagtgtatta tgacccatca
4081 aaagacttaa tagcagaaat acagaagcag gggcaaggcc aatggacata tcaaatttat
4141 caagagccat ttaaaaatct gaaaacagga aagtatgcaa gaatgaaggg tgcccacact
4201 aatgatgtga aacaattaac agaggcagta caaaaaatag ccacagaaag catagtaata
4261 tggggaaaga ctcctaaatt taaattaccc atacaaaagg aaacatggga agcatggtgg
4321 acagagtatt ggcaagccac ctggattcct gagtgggagt ttgtcaatac ccctccctta
4381 gtgaagttat ggtaccagtt agagaaagaa cccataatag gagcagaaac tttctatgta
4441 gatggggcag ccaataggga aactaaatta ggaaaagcag gatatgtaac tgacagagga
4501 agacaaaaag ttgtccccct aacggacaca acaaatcaga agactgagtt acaagcaatt
4561 catctagctt tgcaggattc gggattagaa gtaaacatag tgacagactc acaatatgca
4621 ttgggaatca ttcaagcaca accagataag agtgaatcag agttagtcag tcaaataata
4681 gagcagttaa taaaaaagga aaaagtctac ctggcatggg taccagcaca caaaggaatt
4741 ggaggaaatg aacaagtaga taaattggtc agtgctggaa tcaggaaagt actattttta
4801 gatggaatag ataaggccca agaagaacat gagaaatatc acagtaattg gagagcaatg
4861 gctagtgatt ttaacctacc acctgtagta gcaaaagaaa tagtagccag ctgtgataaa
4921 tgtcagctaa aaggggaagc catgcatgga caagtagact gtagcccagg aatatggcag
4981 ctagattgta cacatttaga aggaaaagtt atcttggtag cagttcatgt agccagtgga
5041 tatatagaag cagaagtaat tccagcagag acagggcaag aaacagcata cttcctctta
5101 aaattagcag gaagatggcc agtaaaaaca gtacatacag acaatggcag caatttcacc
5161 agtactacag ttaaggccgc ctgttggtgg gcggggatca agcaggaatt tggcattccc
5221 tacaatcccc aaagtcaagg agtaatagaa tctatgaata aagaattaaa gaaaattata
5281 ggacaggtaa gagatcaggc tgaacatctt aagacagcag tacaaatggc agtattcatc
5341 cacaatttta aaagaaaagg ggggattggg gggtacagtg caggggaaag aatagtagac
5401 ataatagcaa cagacataca aactaaagaa ttacaaaaac aaattacaaa aattcaaaat
5461 tttcgggttt attacaggga cagcagagat ccagtttgga aaggaccagc aaagctcctc
5521 tggaaaggtg aaggggcagt agtaatacaa gataatagtg acataaaagt agtgccaaga
5581 agaaaagcaa agatcatcag ggattatgga aaacagatgg caggtgatga ttgtgtggca
5641 agtagacagg atgaggatta acacatggaa ttccggagcg gccgcaggag ctttgttcct
5701 tgggttcttg ggagcagcag gaagcactat gggcgcagcg tcaatgacgc tgacggtaca
5761 ggccagacaa ttattgtctg gtatagtgca gcagcagaac aatttgctga gggctattga
5821 ggcgcaacag catctgttgc aactcacagt ctggggcatc aagcagctcc aggcaagaat
5881 cctggctgtg gaaagatacc taaaggatca acagctcctg gggatttggg gttgctctgg
5941 aaaactcatt tgcaccactg ctgtgccttg gaatgctagt tggagtaata aatctctgga
6001 acagatttgg aatcacacga cctggatgga gtgggacaga gaaattaaca attacacaag
6061 cttccgcgga attcacccca ccagtgcagg ctgcctatca gaaagtggtg gctggtgtgg
6121 ctaatgccct ggcccacaag tatcactaag ctcgctttct tgctgtccaa tttctattaa
6181 aggttccttt gttccctaag tccaactact aaactggggg atattatgaa gggccttgag
6241 catctggatt ctgcctaata aaaaacattt attttcattg caatgatgta tttaaattat
6301 ttctgaatat tttactaaaa agggaatgtg ggaggtcagt gcatttaaaa cataaagaaa
6361 tgaagagcta gttcaaacct tgggaaaata cactatatct taaactccat gaaagaaggt
6421 gaggctgcaa acagctaatg cacattggca acagcccctg atgcctatgc cttattcatc
6481 cctcagaaaa ggattcaagt agaggcttga tttggaggtt aaagttttgc tatgctgtat
6541 tttacattac ttattgtttt agctgtcctc atgaatgtct tttcactacc catttgctta
6601 tcctgcatct ctcagccttg actccactca gttctcttgc ttagagatac cacctttccc
6661 ctgaagtgtt ccttccatgt tttacggcga gatggtttct cctcgcctgg ccactcagcc
6721 ttagttgtct ctgttgtctt atagaggtct acttgaagaa ggaaaaacag ggggcatggt
6781 ttgactgtcc tgtgagccct tcttccctgc ctcccccact cacagtgacc cggaatccct
6841 cgacatggca gtctagcact agtgcggccg cagatctgct tcctcgctca ctgactcgct
6901 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
6961 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
7021 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
7081 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
7141 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
7201 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
7261 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
7321 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
7381 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
7441 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
7501 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
7561 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
7621 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
7681 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
7741 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
7801 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
7861 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
7921 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
7981 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
8041 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
8101 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
8161 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
8221 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
8281 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
8341 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
8401 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
8461 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
8521 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
8581 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
8641 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
8701 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
8761 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacggga tcccctgagg
8821 gggcccccat gggctagagg atccggcctc ggcctctgca taaataaaaa aaattagtca
8881 gccatgagc
//
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：pLP1慢病毒包装质粒
别称: pLP-1;pGag/Pol
| 启动子: | CMV |
|---|---|
| 复制子: | pUC |
| 终止子: | β-globin poly(A) |
| 质粒分类: | 病毒系列，慢病毒包装载体 |
| 质粒大小: | 8889bp |
| 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
PLP1 (Proteolipid Protein 1) is a Protein Coding gene. Diseases associated with PLP1 include pelizaeus-merzbacher disease and spastic paraplegia 2, x-linked. Among its related pathways are Glial Cell Differentiation and SIDS Susceptibility Pathways. GO annotations related to this gene include structural molecule activity and structural constituent of myelin sheath. An important paralog of this gene is GPM6B
质粒图谱
质粒序列
LOCUS Exported 8889 bp ds-DNA circular SYN 27-AUG-2016
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS Untitled 32
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8889)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Saturday, August 27, 2016 from SnapGene Viewer 3.1.4
http://www.miaolingbio.com
FEATURES Location/Qualifiers
source 1..8889
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 79..458
/note="CMV enhancer"
/note="human cytomegalovirus immediate early enhancer"
promoter 459..662
/note="CMV promoter"
/note="human cytomegalovirus (CMV) immediate early
promoter"
intron 796..1271
/note="beta-globin intron"
/note="internally truncated intron from human beta-globin"
CDS 1355..2857
/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 2650..5661
/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 5346..5463
/note="cPPT/CTS"
/note="central polypurine tract and central termination
sequence of HIV-1"
misc_feature 5686..5919
/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 6150..6544
/note="beta-globin poly(A)"
/note="human beta-globin polyadenylation signal"
rep_origin complement(7054..7642)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(7813..8673)
/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(8674..8778)
/gene="bla"
/note="AmpR promoter"
ORIGIN
1 ttggcccatt gcatacgttg tatccatatc ataatatgta catttatatt ggctcatgtc
61 caacattacc gccatgttga cattgattat tgactagtta ttaatagtaa tcaattacgg
121 ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc
181 cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca
241 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg
301 cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg
361 acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt
421 ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca
481 tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg
541 tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact
601 ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag
661 ctcgtttagt gaaccgtcag atcgcctgga gacgccatcc acgctgtttt gacctccata
721 gaagacaccg ggaccgatcc agcctcccct cgaagcttac atgtggtacc gagctcggat
781 cctgagaact tcagggtgag tctatgggac ccttgatgtt ttctttcccc ttcttttcta
841 tggttaagtt catgtcatag gaaggggaga agtaacaggg tacacatatt gaccaaatca
901 gggtaatttt gcatttgtaa ttttaaaaaa tgctttcttc ttttaatata cttttttgtt
961 tatcttattt ctaatacttt ccctaatctc tttctttcag ggcaataatg atacaatgta
1021 tcatgcctct ttgcaccatt ctaaagaata acagtgataa tttctgggtt aaggcaatag
1081 caatatttct gcatataaat atttctgcat ataaattgta actgatgtaa gaggtttcat
1141 attgctaata gcagctacaa tccagctacc attctgcttt tattttatgg ttgggataag
1201 gctggattat tctgagtcca agctaggccc ttttgctaat catgttcata cctcttatct
1261 tcctcccaca gctcctgggc aacgtgctgg tctgtgtgct ggcccatcac tttggcaaag
1321 cacgtgagat ctgaattcga gatctgccgc cgccatgggt gcgagagcgt cagtattaag
1381 cgggggagaa ttagatcgat gggaaaaaat tcggttaagg ccagggggaa agaaaaaata
1441 taaattaaaa catatagtat gggcaagcag ggagctagaa cgattcgcag ttaatcctgg
1501 cctgttagaa acatcagaag gctgtagaca aatactggga cagctacaac catcccttca
1561 gacaggatca gaagaactta gatcattata taatacagta gcaaccctct attgtgtgca
1621 tcaaaggata gagataaaag acaccaagga agctttagac aagatagagg aagagcaaaa
1681 caaaagtaag aaaaaagcac agcaagcagc agctgacaca ggacacagca atcaggtcag
1741 ccaaaattac cctatagtgc agaacatcca ggggcaaatg gtacatcagg ccatatcacc
1801 tagaacttta aatgcatggg taaaagtagt agaagagaag gctttcagcc cagaagtgat
1861 acccatgttt tcagcattat cagaaggagc caccccacaa gatttaaaca ccatgctaaa
1921 cacagtgggg ggacatcaag cagccatgca aatgttaaaa gagaccatca atgaggaagc
1981 tgcagaatgg gatagagtgc atccagtgca tgcagggcct attgcaccag gccagatgag
2041 agaaccaagg ggaagtgaca tagcaggaac tactagtacc cttcaggaac aaataggatg
2101 gatgacacat aatccaccta tcccagtagg agaaatctat aaaagatgga taatcctggg
2161 attaaataaa atagtaagaa tgtatagccc taccagcatt ctggacataa gacaaggacc
2221 aaaggaaccc tttagagact atgtagaccg attctataaa actctaagag ccgagcaagc
2281 ttcacaagag gtaaaaaatt ggatgacaga aaccttgttg gtccaaaatg cgaacccaga
2341 ttgtaagact attttaaaag cattgggacc aggagcgaca ctagaagaaa tgatgacagc
2401 atgtcaggga gtggggggac ccggccataa agcaagagtt ttggctgaag caatgagcca
2461 agtaacaaat ccagctacca taatgataca gaaaggcaat tttaggaacc aaagaaagac
2521 tgttaagtgt ttcaattgtg gcaaagaagg gcacatagcc aaaaattgca gggcccctag
2581 gaaaaagggc tgttggaaat gtggaaagga aggacaccaa atgaaagatt gtactgagag
2641 acaggctaat tttttaggga agatctggcc ttcccacaag ggaaggccag ggaattttct
2701 tcagagcaga ccagagccaa cagccccacc agaagagagc ttcaggtttg gggaagagac
2761 aacaactccc tctcagaagc aggagccgat agacaaggaa ctgtatcctt tagcttccct
2821 cagatcactc tttggcagcg acccctcgtc acaataaaga taggggggca attaaaggaa
2881 gctctattag atacaggagc agatgataca gtattagaag aaatgaattt gccaggaaga
2941 tggaaaccaa aaatgatagg gggaattgga ggttttatca aagtaagaca gtatgatcag
3001 atactcatag aaatctgcgg acataaagct ataggtacag tattagtagg acctacacct
3061 gtcaacataa ttggaagaaa tctgttgact cagattggct gcactttaaa ttttcccatt
3121 agtcctattg agactgtacc agtaaaatta aagccaggaa tggatggccc aaaagttaaa
3181 caatggccat tgacagaaga aaaaataaaa gcattagtag aaatttgtac agaaatggaa
3241 aaggaaggaa aaatttcaaa aattgggcct gaaaatccat acaatactcc agtatttgcc
3301 ataaagaaaa aagacagtac taaatggaga aaattagtag atttcagaga acttaataag
3361 agaactcaag atttctggga agttcaatta ggaataccac atcctgcagg gttaaaacag
3421 aaaaaatcag taacagtact ggatgtgggc gatgcatatt tttcagttcc cttagataaa
3481 gacttcagga agtatactgc atttaccata cctagtataa acaatgagac accagggatt
3541 agatatcagt acaatgtgct tccacaggga tggaaaggat caccagcaat attccagtgt
3601 agcatgacaa aaatcttaga gccttttaga aaacaaaatc cagacatagt catctatcaa
3661 tacatggatg atttgtatgt aggatctgac ttagaaatag ggcagcatag aacaaaaata
3721 gaggaactga gacaacatct gttgaggtgg ggatttacca caccagacaa aaaacatcag
3781 aaagaacctc cattcctttg gatgggttat gaactccatc ctgataaatg gacagtacag
3841 cctatagtgc tgccagaaaa ggacagctgg actgtcaatg acatacagaa attagtggga
3901 aaattgaatt gggcaagtca gatttatgca gggattaaag taaggcaatt atgtaaactt
3961 cttaggggaa ccaaagcact aacagaagta gtaccactaa cagaagaagc agagctagaa
4021 ctggcagaaa acagggagat tctaaaagaa ccggtacatg gagtgtatta tgacccatca
4081 aaagacttaa tagcagaaat acagaagcag gggcaaggcc aatggacata tcaaatttat
4141 caagagccat ttaaaaatct gaaaacagga aagtatgcaa gaatgaaggg tgcccacact
4201 aatgatgtga aacaattaac agaggcagta caaaaaatag ccacagaaag catagtaata
4261 tggggaaaga ctcctaaatt taaattaccc atacaaaagg aaacatggga agcatggtgg
4321 acagagtatt ggcaagccac ctggattcct gagtgggagt ttgtcaatac ccctccctta
4381 gtgaagttat ggtaccagtt agagaaagaa cccataatag gagcagaaac tttctatgta
4441 gatggggcag ccaataggga aactaaatta ggaaaagcag gatatgtaac tgacagagga
4501 agacaaaaag ttgtccccct aacggacaca acaaatcaga agactgagtt acaagcaatt
4561 catctagctt tgcaggattc gggattagaa gtaaacatag tgacagactc acaatatgca
4621 ttgggaatca ttcaagcaca accagataag agtgaatcag agttagtcag tcaaataata
4681 gagcagttaa taaaaaagga aaaagtctac ctggcatggg taccagcaca caaaggaatt
4741 ggaggaaatg aacaagtaga taaattggtc agtgctggaa tcaggaaagt actattttta
4801 gatggaatag ataaggccca agaagaacat gagaaatatc acagtaattg gagagcaatg
4861 gctagtgatt ttaacctacc acctgtagta gcaaaagaaa tagtagccag ctgtgataaa
4921 tgtcagctaa aaggggaagc catgcatgga caagtagact gtagcccagg aatatggcag
4981 ctagattgta cacatttaga aggaaaagtt atcttggtag cagttcatgt agccagtgga
5041 tatatagaag cagaagtaat tccagcagag acagggcaag aaacagcata cttcctctta
5101 aaattagcag gaagatggcc agtaaaaaca gtacatacag acaatggcag caatttcacc
5161 agtactacag ttaaggccgc ctgttggtgg gcggggatca agcaggaatt tggcattccc
5221 tacaatcccc aaagtcaagg agtaatagaa tctatgaata aagaattaaa gaaaattata
5281 ggacaggtaa gagatcaggc tgaacatctt aagacagcag tacaaatggc agtattcatc
5341 cacaatttta aaagaaaagg ggggattggg gggtacagtg caggggaaag aatagtagac
5401 ataatagcaa cagacataca aactaaagaa ttacaaaaac aaattacaaa aattcaaaat
5461 tttcgggttt attacaggga cagcagagat ccagtttgga aaggaccagc aaagctcctc
5521 tggaaaggtg aaggggcagt agtaatacaa gataatagtg acataaaagt agtgccaaga
5581 agaaaagcaa agatcatcag ggattatgga aaacagatgg caggtgatga ttgtgtggca
5641 agtagacagg atgaggatta acacatggaa ttccggagcg gccgcaggag ctttgttcct
5701 tgggttcttg ggagcagcag gaagcactat gggcgcagcg tcaatgacgc tgacggtaca
5761 ggccagacaa ttattgtctg gtatagtgca gcagcagaac aatttgctga gggctattga
5821 ggcgcaacag catctgttgc aactcacagt ctggggcatc aagcagctcc aggcaagaat
5881 cctggctgtg gaaagatacc taaaggatca acagctcctg gggatttggg gttgctctgg
5941 aaaactcatt tgcaccactg ctgtgccttg gaatgctagt tggagtaata aatctctgga
6001 acagatttgg aatcacacga cctggatgga gtgggacaga gaaattaaca attacacaag
6061 cttccgcgga attcacccca ccagtgcagg ctgcctatca gaaagtggtg gctggtgtgg
6121 ctaatgccct ggcccacaag tatcactaag ctcgctttct tgctgtccaa tttctattaa
6181 aggttccttt gttccctaag tccaactact aaactggggg atattatgaa gggccttgag
6241 catctggatt ctgcctaata aaaaacattt attttcattg caatgatgta tttaaattat
6301 ttctgaatat tttactaaaa agggaatgtg ggaggtcagt gcatttaaaa cataaagaaa
6361 tgaagagcta gttcaaacct tgggaaaata cactatatct taaactccat gaaagaaggt
6421 gaggctgcaa acagctaatg cacattggca acagcccctg atgcctatgc cttattcatc
6481 cctcagaaaa ggattcaagt agaggcttga tttggaggtt aaagttttgc tatgctgtat
6541 tttacattac ttattgtttt agctgtcctc atgaatgtct tttcactacc catttgctta
6601 tcctgcatct ctcagccttg actccactca gttctcttgc ttagagatac cacctttccc
6661 ctgaagtgtt ccttccatgt tttacggcga gatggtttct cctcgcctgg ccactcagcc
6721 ttagttgtct ctgttgtctt atagaggtct acttgaagaa ggaaaaacag ggggcatggt
6781 ttgactgtcc tgtgagccct tcttccctgc ctcccccact cacagtgacc cggaatccct
6841 cgacatggca gtctagcact agtgcggccg cagatctgct tcctcgctca ctgactcgct
6901 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
6961 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
7021 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
7081 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
7141 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
7201 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
7261 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
7321 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
7381 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
7441 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
7501 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
7561 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
7621 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
7681 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
7741 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
7801 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
7861 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
7921 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
7981 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
8041 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
8101 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
8161 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
8221 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
8281 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
8341 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
8401 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
8461 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
8521 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
8581 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
8641 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
8701 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
8761 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacggga tcccctgagg
8821 gggcccccat gggctagagg atccggcctc ggcctctgca taaataaaaa aaattagtca
8881 gccatgagc
//
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
| P1778/pENTR223-IWS1人源基因质粒 |
| P1779/pENTR223-LINC00152人源基因质粒 |
| P1791/pENTR223-NBR2人源基因质粒 |
| P1792/pENTR223-ZNF706人源基因质粒 |
| P1793/pENTR223-NICE-5人源基因质粒 |
| P1794/pENTR223-ARL17B人源基因质粒 |
| P1795/pENTR223-COX6A2人源基因质粒 |
| P1859/pENTR223-MLLT11人源基因质粒 |
| P1796/pENTR223-MLLT11-G262C人源基因质粒 |
| P0609/pENTR223-FNDC5人源基因质粒 |
| P0611/pENTR223-RLN2人源基因质粒 |
| P0531/pENTR223-LGALS13人源基因质粒 |
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pLP1慢病毒包装质粒
￥100 - 1000
