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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：
2年
- 英文名：
Plasmid#83842
- 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：pLL3.7m-mTurquoise2-SLBP(18-126)-IRES-H1-mMaroon1人源基因质粒
别称: Plasmid#83842
启动子：CMV
复制子: pUC
原核抗性: Amp
克隆菌株: DH5a
培养条件: 37度
质粒属性
载体宿主：哺乳细胞
载体用途：蛋白表达
基因种属：人
基因类型：ORF
原核抗性：Amp
真核抗性：
荧光蛋白：
质粒简介：详询
质粒图谱
质粒序列
LOCUS Exported 9431 bp ds-DNA circular SYN 27-SEP-2017
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9431)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Sep 27, 2017 from MiaoLingPlasmid
http://www.miaolingbio.com
FEATURES Location/Qualifiers
source 1..9431
/organism="synthetic DNA construct"
/mol_type="other DNA"
misc_feature 101..562
/label=IRES
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
CDS 556..1902
/codon_start=1
/label=H1-mMaroon1
/translation="MIMATEFMTENSTSAPAAKPKRAKASKKSTDHPKYSDMIVAAIQA
EKNRAGSSRQSIQKYIKSHYKVGENADSQIKLSIKRLVTTGVLKQTKGVGASGSFRLAK
GDEPKRSVAFKKTKKEVKKVATPKKAAKPKKAASKAPSKKPKATPVKKAKKKPAATPKK
AKKPKVVKVKPVKASKPKKAKTVKPKAKSSAKRGSKKKSGLRSGGSGGMVSKGEELIKE
NMHTKLYLTGTVNNHYFECTAEGEGKPYEGTQTNRIKVVRGGPLPFAFDILAPCFMYGS
KTFINHPPDIPDYFKQSFPEGFTWERTTVYEDGGTLTATQDTSLQDGCLIYNVQVRGEN
FPSNGPVMQKKTLGWEASTETLYPADGSLEGRLDWALKLVGGGHLHCRLETTYRSKKPA
KNLKMPGVYFIDRRLERIKEADNETYVEQHEVAVARYCDLPSKLGHKLN"
protein_bind 1930..1963
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT)."
misc_feature 2019..2607
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
LTR 2849..3029
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
rep_origin complement(3091..3679)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(3850..4710)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(4711..4815)
/gene="bla"
/label=AmpR promoter
enhancer 4906..5209
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 5211..5409
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
LTR 5427..5607
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 5654..5779
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 6276..6509
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
misc_feature 7036..7153
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
protein_bind 7599..7632
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT)."
enhancer 7748..8051
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 8052..8255
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
CDS 8291..9007
/codon_start=1
/product="enhanced monomeric variant of CFP (Goedhart et
al., 2012)"
/label=mTurquoise2
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYFSDNVYITADKQKNGIK
ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
misc_feature 9008..9400
/label=SLBP(18-126)
ORIGIN
1 cccccccccc taacgttact ggccgaagcc gcttggaata aggccggtgt gcgtttgtct
61 atatgttatt ttccaccata ttgccgtctt ttggcaatgt gagggcccgg aaacctggcc
121 ctgtcttctt gacgagcatt cctaggggtc tttcccctct cgccaaagga atgcaaggtc
181 tgttgaatgt cgtgaaggaa gcagttcctc tggaagcttc ttgaagacaa acaacgtctg
241 tagcgaccct ttgcaggcag cggaaccccc cacctggcga caggtgcctc tgcggccaaa
301 agccacgtgt ataagataca cctgcaaagg cggcacaacc ccagtgccac gttgtgagtt
361 ggatagttgt ggaaagagtc aaatggctct cctcaagcgt attcaacaag gggctgaagg
421 atgcccagaa ggtaccccat tgtatgggat ctgatctggg gcctcggtgc acatgcttta
481 catgtgttta gtcgaggtta aaaaacgtct aggccccccg aaccacgggg acgtggtttt
541 cctttgaaaa acacgatgat aatggccaca gaattcatga ccgagaactc cacctccgcc
601 ccggcggcga agcccaaacg ggccaaggct tccaagaagt ccacggacca ccccaagtat
661 tcagacatga tcgtggctgc tatccaggca gagaagaacc gtgccggctc ctcgcgccag
721 tccatccaaa agtatatcaa gagccactac aaggtgggtg agaacgccga ctcccagatc
781 aagttgtcca tcaagcgcct agtgaccacc ggtgttctca agcaaaccaa aggggtgggc
841 gcctcggggt ccttcaggct ggccaagggc gatgagccca aaaggtcggt ggctttcaag
901 aagaccaaga aggaagtcaa gaaagtggcc actccaaaga aggcagccaa gcccaagaag
961 gctgcctcca aagccccaag caagaaaccc aaagccaccc ctgtcaagaa ggccaagaag
1021 aagccggctg ccacgcccaa gaaagccaaa aagcccaagg ttgtcaaagt caaaccagtc
1081 aaggcctcca aacccaagaa ggccaaaacc gtgaagccca aagccaagtc gagtgccaag
1141 agggggagca agaagaagtc cggactcaga tctggaggct ccggaggcat ggtgagcaag
1201 ggcgaggagc tgatcaagga gaacatgcac accaagctgt acctgacagg caccgtgaac
1261 aaccactact tcgaatgcac cgccgaaggg gagggcaagc cctacgaggg cacccagacc
1321 aacaggatca aggtggtgcg cggaggcccc ctgccgttcg cattcgacat cctggccccc
1381 tgctttatgt acgggagcaa gaccttcatt aaccatccac ctgacattcc cgattatttt
1441 aagcagtcct tccctgaggg cttcacatgg gagagaacca ccgtttacga agacgggggc
1501 acactcactg ctacacagga caccagcctc caggacggct gcctgatcta caacgtccaa
1561 gtcagagggg agaacttccc atccaacggc cctgtgatgc agaagaaaac actcggctgg
1621 gaggccagta ccgagaccct gtaccccgct gacggcagcc tggaaggcag actggactgg
1681 gccctgaagc tcgtgggcgg gggccacctg cactgcagac tggaaaccac atacagatcc
1741 aagaaacccg caaagaacct caagatgccc ggcgtctatt ttattgacag gagactggaa
1801 agaatcaagg aggccgacaa tgagacctac gtcgagcagc acgaggtggc tgtggccaga
1861 tactgcgacc tccctagcaa actggggcac aaacttaatt gagtttaaac gaattcgtcg
1921 agggacctaa taacttcgta tagcatacat tatacgaagt tatacatgtt taagggttcc
1981 ggttccacta ggtacaattc gatatcaagc ttatcgataa tcaacctctg gattacaaaa
2041 tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg
2101 ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct
2161 tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg
2221 gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct
2281 gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg
2341 ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg
2401 tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc acctggattc
2461 tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc
2521 gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc
2581 ggatctccct ttgggccgcc tccccgcatc gataccgtcg acctcgatcg agacctagaa
2641 aaacatggag caatcacaag tagcaataca gcagctacca atgctgattg tgcctggcta
2701 gaagcacaag aggaggagga ggtgggtttt ccagtcacac ctcaggtacc tttaagacca
2761 atgacttaca aggcagctga cgcgtgattt taaaagaaaa ggggggactg gaagggctaa
2821 ttcactccca ctgctttttg cctgtactgg gtctctctgg ttagaccaga tctgagcctg
2881 ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt
2941 gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc
3001 cttttagtca gtgtggaaaa tctctagcag catgtgagca aaaggccagc aaaaggccag
3061 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
3121 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
3181 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
3241 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
3301 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
3361 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
3421 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
3481 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
3541 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
3601 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
3661 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
3721 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
3781 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
3841 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
3901 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
3961 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
4021 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
4081 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
4141 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
4201 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
4261 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
4321 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
4381 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
4441 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
4501 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
4561 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
4621 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
4681 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
4741 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
4801 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacactagtt attaatagta
4861 atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac
4921 ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac
4981 gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt
5041 acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat
5101 tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga
5161 ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt
5221 ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca
5281 ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg
5341 tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta
5401 tataagcagc gcgttttgcc tgtactgggt ctctctggtt agaccagatc tgagcctggg
5461 agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc
5521 ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct
5581 tttagtcagt gtggaaaatc tctagcagtg gcgcccgaac agggacttga aagcgaaagg
5641 gaaaccagag gagctctctc gacgcaggac tcggcttgct gaagcgcgca cggcaagagg
5701 cgaggggcgg cgactggtga gtacgccaaa aattttgact agcggaggct agaaggagag
5761 agatgggtgc gagagcgtca gtattaagcg ggggagaatt agatcgcgat gggaaaaaat
5821 tcggttaagg ccagggggaa agaaaaaata taaattaaaa catatagtat gggcaagcag
5881 ggagctagaa cgattcgcag ttaatcctgg cctgttagaa acatcagaag gctgtagaca
5941 aatactggga cagctacaac catcccttca gacaggatca gaagaactta gatcattata
6001 taatacagta gcaaccctct attgtgtgca tcaaaggata gagataaaag acaccaagga
6061 agctttagac aagatagagg aagagcaaaa caaaagtaag accaccgcac agcaagcggc
6121 cggccgctga tcttcagacc tggaggagga gatatgaggg acaattggag aagtgaatta
6181 tataaatata aagtagtaaa aattgaacca ttaggagtag cacccaccaa ggcaaagaga
6241 agagtggtgc agagagaaaa aagagcagtg ggaataggag ctttgttcct tgggttcttg
6301 ggagcagcag gaagcactat gggcgcagcg tcaatgacgc tgacggtaca ggccagacaa
6361 ttattgtctg gtatagtgca gcagcagaac aatttgctga gggctattga ggcgcaacag
6421 catctgttgc aactcacagt ctggggcatc aagcagctcc aggcaagaat cctggctgtg
6481 gaaagatacc taaaggatca acagctcctg gggatttggg gttgctctgg aaaactcatt
6541 tgcaccactg ctgtgccttg gaatgctagt tggagtaata aatctctgga acagatttgg
6601 aatcacacga cctggatgga gtgggacaga gaaattaaca attacacaag cttaatacac
6661 tccttaattg aagaatcgca aaaccagcaa gaaaagaatg aacaagaatt attggaatta
6721 gataaatggg caagtttgtg gaattggttt aacataacaa attggctgtg gtatataaaa
6781 ttattcataa tgatagtagg aggcttggta ggtttaagaa tagtttttgc tgtactttct
6841 atagtgaata gagttaggca gggatattca ccattatcgt ttcagaccca cctcccaacc
6901 ccgaggggac ccgacaggcc cgaaggaata gaagaagaag gtggagagag agacagagac
6961 agatccattc gattagtgaa cggatcggca ctgcgtgcgc caattctgca gacaaatggc
7021 agtattcatc cacaatttta aaagaaaagg ggggattggg gggtacagtg caggggaaag
7081 aatagtagac ataatagcaa cagacataca aactaaagaa ttacaaaaac aaattacaaa
7141 aattcaaaat tttcgggttt attacaggga cagcagagat ccagtttggt tagtaccggg
7201 cccgctctag agatccgacg cgccatctct aggcccgcgc cggccccctc gcacagactt
7261 gtgggagaag ctcggctact cccctgcccc ggttaatttg catataatat ttcctagtaa
7321 ctatagaggc ttaatgtgcg ataaaagaca gataatctgt tctttttaat actagctaca
7381 ttttacatga taggcttgga tttctataag agatacaaat actaaattat tattttaaaa
7441 aacagcacaa aaggaaactc accctaactg taaagtaatt gtgtgttttg agactataaa
7501 tatcccttgg agaaaagcct tgttaacgcg cggtgaccct cgaggtcgac ggtatcgata
7561 agctcgcttc acgagattcc agcaggtcga gggacctaat aacttcgtat agcatacatt
7621 atacgaagtt atattaaggg ttccaagctt aagcggccgc gtggataacc gtattaccgc
7681 catgcattag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
7741 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
7801 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
7861 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
7921 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
7981 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
8041 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
8101 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
8161 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
8221 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct
8281 agcaccaggt atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt
8341 cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga
8401 tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc
8461 ctggcccacc ctcgtgacca ccctgagctg gggcgtgcag tgcttcgccc gctaccccga
8521 ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg
8581 caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg
8641 cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat
8701 cctggggcac aagctggagt acaactactt cagcgacaac gtctatatca ccgccgacaa
8761 gcagaagaac ggcatcaagg ccaacttcaa gatccgccac aacatcgagg acggcggcgt
8821 gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc
8881 cgacaaccac tacctgagca cccagtccaa gctgagcaaa gaccccaacg agaagcgcga
8941 tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct
9001 gtacaagtac tcaatgttgc cttccctggg aggataccca tcacactggc gaccactcga
9061 ggccagcccg ccgtcccccg cgcgatggag cctgggacgg aagcgcagag ccgacggcag
9121 gcgctggagg cccgaagacg ccgaggaggc agagcaccgc ggcgccgagc gcagacccga
9181 gagctttacc actcctgaag gccctaaacc ccgttccaga tgctctgact gggcaagtgc
9241 agttgaagaa gatgaaatga ggaccagagt taacaaagaa atggcaagat ataaaaggaa
9301 actcctcatc aatgactttg gaagagagag aaaatcatca tcaggaagtt ctgattcaaa
9361 ggagtctatg tctactgtgc cggctgacga tcgaggatga agaaggatct tagcttaagg
9421 cccctctccc t
//
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：pLL3.7m-mTurquoise2-SLBP(18-126)-IRES-H1-mMaroon1人源基因质粒
别称: Plasmid#83842
启动子：CMV
复制子: pUC
原核抗性: Amp
克隆菌株: DH5a
培养条件: 37度
质粒属性
载体宿主：哺乳细胞
载体用途：蛋白表达
基因种属：人
基因类型：ORF
原核抗性：Amp
真核抗性：
荧光蛋白：
质粒简介：详询
质粒图谱
质粒序列
LOCUS Exported 9431 bp ds-DNA circular SYN 27-SEP-2017
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9431)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Sep 27, 2017 from MiaoLingPlasmid
http://www.miaolingbio.com
FEATURES Location/Qualifiers
source 1..9431
/organism="synthetic DNA construct"
/mol_type="other DNA"
misc_feature 101..562
/label=IRES
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
CDS 556..1902
/codon_start=1
/label=H1-mMaroon1
/translation="MIMATEFMTENSTSAPAAKPKRAKASKKSTDHPKYSDMIVAAIQA
EKNRAGSSRQSIQKYIKSHYKVGENADSQIKLSIKRLVTTGVLKQTKGVGASGSFRLAK
GDEPKRSVAFKKTKKEVKKVATPKKAAKPKKAASKAPSKKPKATPVKKAKKKPAATPKK
AKKPKVVKVKPVKASKPKKAKTVKPKAKSSAKRGSKKKSGLRSGGSGGMVSKGEELIKE
NMHTKLYLTGTVNNHYFECTAEGEGKPYEGTQTNRIKVVRGGPLPFAFDILAPCFMYGS
KTFINHPPDIPDYFKQSFPEGFTWERTTVYEDGGTLTATQDTSLQDGCLIYNVQVRGEN
FPSNGPVMQKKTLGWEASTETLYPADGSLEGRLDWALKLVGGGHLHCRLETTYRSKKPA
KNLKMPGVYFIDRRLERIKEADNETYVEQHEVAVARYCDLPSKLGHKLN"
protein_bind 1930..1963
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT)."
misc_feature 2019..2607
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
LTR 2849..3029
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
rep_origin complement(3091..3679)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(3850..4710)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(4711..4815)
/gene="bla"
/label=AmpR promoter
enhancer 4906..5209
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 5211..5409
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
LTR 5427..5607
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 5654..5779
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 6276..6509
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
misc_feature 7036..7153
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
protein_bind 7599..7632
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT)."
enhancer 7748..8051
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 8052..8255
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
CDS 8291..9007
/codon_start=1
/product="enhanced monomeric variant of CFP (Goedhart et
al., 2012)"
/label=mTurquoise2
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYFSDNVYITADKQKNGIK
ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
misc_feature 9008..9400
/label=SLBP(18-126)
ORIGIN
1 cccccccccc taacgttact ggccgaagcc gcttggaata aggccggtgt gcgtttgtct
61 atatgttatt ttccaccata ttgccgtctt ttggcaatgt gagggcccgg aaacctggcc
121 ctgtcttctt gacgagcatt cctaggggtc tttcccctct cgccaaagga atgcaaggtc
181 tgttgaatgt cgtgaaggaa gcagttcctc tggaagcttc ttgaagacaa acaacgtctg
241 tagcgaccct ttgcaggcag cggaaccccc cacctggcga caggtgcctc tgcggccaaa
301 agccacgtgt ataagataca cctgcaaagg cggcacaacc ccagtgccac gttgtgagtt
361 ggatagttgt ggaaagagtc aaatggctct cctcaagcgt attcaacaag gggctgaagg
421 atgcccagaa ggtaccccat tgtatgggat ctgatctggg gcctcggtgc acatgcttta
481 catgtgttta gtcgaggtta aaaaacgtct aggccccccg aaccacgggg acgtggtttt
541 cctttgaaaa acacgatgat aatggccaca gaattcatga ccgagaactc cacctccgcc
601 ccggcggcga agcccaaacg ggccaaggct tccaagaagt ccacggacca ccccaagtat
661 tcagacatga tcgtggctgc tatccaggca gagaagaacc gtgccggctc ctcgcgccag
721 tccatccaaa agtatatcaa gagccactac aaggtgggtg agaacgccga ctcccagatc
781 aagttgtcca tcaagcgcct agtgaccacc ggtgttctca agcaaaccaa aggggtgggc
841 gcctcggggt ccttcaggct ggccaagggc gatgagccca aaaggtcggt ggctttcaag
901 aagaccaaga aggaagtcaa gaaagtggcc actccaaaga aggcagccaa gcccaagaag
961 gctgcctcca aagccccaag caagaaaccc aaagccaccc ctgtcaagaa ggccaagaag
1021 aagccggctg ccacgcccaa gaaagccaaa aagcccaagg ttgtcaaagt caaaccagtc
1081 aaggcctcca aacccaagaa ggccaaaacc gtgaagccca aagccaagtc gagtgccaag
1141 agggggagca agaagaagtc cggactcaga tctggaggct ccggaggcat ggtgagcaag
1201 ggcgaggagc tgatcaagga gaacatgcac accaagctgt acctgacagg caccgtgaac
1261 aaccactact tcgaatgcac cgccgaaggg gagggcaagc cctacgaggg cacccagacc
1321 aacaggatca aggtggtgcg cggaggcccc ctgccgttcg cattcgacat cctggccccc
1381 tgctttatgt acgggagcaa gaccttcatt aaccatccac ctgacattcc cgattatttt
1441 aagcagtcct tccctgaggg cttcacatgg gagagaacca ccgtttacga agacgggggc
1501 acactcactg ctacacagga caccagcctc caggacggct gcctgatcta caacgtccaa
1561 gtcagagggg agaacttccc atccaacggc cctgtgatgc agaagaaaac actcggctgg
1621 gaggccagta ccgagaccct gtaccccgct gacggcagcc tggaaggcag actggactgg
1681 gccctgaagc tcgtgggcgg gggccacctg cactgcagac tggaaaccac atacagatcc
1741 aagaaacccg caaagaacct caagatgccc ggcgtctatt ttattgacag gagactggaa
1801 agaatcaagg aggccgacaa tgagacctac gtcgagcagc acgaggtggc tgtggccaga
1861 tactgcgacc tccctagcaa actggggcac aaacttaatt gagtttaaac gaattcgtcg
1921 agggacctaa taacttcgta tagcatacat tatacgaagt tatacatgtt taagggttcc
1981 ggttccacta ggtacaattc gatatcaagc ttatcgataa tcaacctctg gattacaaaa
2041 tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg
2101 ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct
2161 tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg
2221 gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct
2281 gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg
2341 ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg
2401 tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc acctggattc
2461 tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc
2521 gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc
2581 ggatctccct ttgggccgcc tccccgcatc gataccgtcg acctcgatcg agacctagaa
2641 aaacatggag caatcacaag tagcaataca gcagctacca atgctgattg tgcctggcta
2701 gaagcacaag aggaggagga ggtgggtttt ccagtcacac ctcaggtacc tttaagacca
2761 atgacttaca aggcagctga cgcgtgattt taaaagaaaa ggggggactg gaagggctaa
2821 ttcactccca ctgctttttg cctgtactgg gtctctctgg ttagaccaga tctgagcctg
2881 ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt
2941 gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc
3001 cttttagtca gtgtggaaaa tctctagcag catgtgagca aaaggccagc aaaaggccag
3061 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
3121 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
3181 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
3241 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
3301 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
3361 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
3421 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
3481 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
3541 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
3601 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
3661 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
3721 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
3781 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
3841 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
3901 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
3961 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
4021 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
4081 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
4141 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
4201 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
4261 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
4321 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
4381 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
4441 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
4501 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
4561 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
4621 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
4681 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
4741 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
4801 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacactagtt attaatagta
4861 atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac
4921 ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac
4981 gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt
5041 acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat
5101 tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga
5161 ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt
5221 ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca
5281 ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg
5341 tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta
5401 tataagcagc gcgttttgcc tgtactgggt ctctctggtt agaccagatc tgagcctggg
5461 agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc
5521 ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct
5581 tttagtcagt gtggaaaatc tctagcagtg gcgcccgaac agggacttga aagcgaaagg
5641 gaaaccagag gagctctctc gacgcaggac tcggcttgct gaagcgcgca cggcaagagg
5701 cgaggggcgg cgactggtga gtacgccaaa aattttgact agcggaggct agaaggagag
5761 agatgggtgc gagagcgtca gtattaagcg ggggagaatt agatcgcgat gggaaaaaat
5821 tcggttaagg ccagggggaa agaaaaaata taaattaaaa catatagtat gggcaagcag
5881 ggagctagaa cgattcgcag ttaatcctgg cctgttagaa acatcagaag gctgtagaca
5941 aatactggga cagctacaac catcccttca gacaggatca gaagaactta gatcattata
6001 taatacagta gcaaccctct attgtgtgca tcaaaggata gagataaaag acaccaagga
6061 agctttagac aagatagagg aagagcaaaa caaaagtaag accaccgcac agcaagcggc
6121 cggccgctga tcttcagacc tggaggagga gatatgaggg acaattggag aagtgaatta
6181 tataaatata aagtagtaaa aattgaacca ttaggagtag cacccaccaa ggcaaagaga
6241 agagtggtgc agagagaaaa aagagcagtg ggaataggag ctttgttcct tgggttcttg
6301 ggagcagcag gaagcactat gggcgcagcg tcaatgacgc tgacggtaca ggccagacaa
6361 ttattgtctg gtatagtgca gcagcagaac aatttgctga gggctattga ggcgcaacag
6421 catctgttgc aactcacagt ctggggcatc aagcagctcc aggcaagaat cctggctgtg
6481 gaaagatacc taaaggatca acagctcctg gggatttggg gttgctctgg aaaactcatt
6541 tgcaccactg ctgtgccttg gaatgctagt tggagtaata aatctctgga acagatttgg
6601 aatcacacga cctggatgga gtgggacaga gaaattaaca attacacaag cttaatacac
6661 tccttaattg aagaatcgca aaaccagcaa gaaaagaatg aacaagaatt attggaatta
6721 gataaatggg caagtttgtg gaattggttt aacataacaa attggctgtg gtatataaaa
6781 ttattcataa tgatagtagg aggcttggta ggtttaagaa tagtttttgc tgtactttct
6841 atagtgaata gagttaggca gggatattca ccattatcgt ttcagaccca cctcccaacc
6901 ccgaggggac ccgacaggcc cgaaggaata gaagaagaag gtggagagag agacagagac
6961 agatccattc gattagtgaa cggatcggca ctgcgtgcgc caattctgca gacaaatggc
7021 agtattcatc cacaatttta aaagaaaagg ggggattggg gggtacagtg caggggaaag
7081 aatagtagac ataatagcaa cagacataca aactaaagaa ttacaaaaac aaattacaaa
7141 aattcaaaat tttcgggttt attacaggga cagcagagat ccagtttggt tagtaccggg
7201 cccgctctag agatccgacg cgccatctct aggcccgcgc cggccccctc gcacagactt
7261 gtgggagaag ctcggctact cccctgcccc ggttaatttg catataatat ttcctagtaa
7321 ctatagaggc ttaatgtgcg ataaaagaca gataatctgt tctttttaat actagctaca
7381 ttttacatga taggcttgga tttctataag agatacaaat actaaattat tattttaaaa
7441 aacagcacaa aaggaaactc accctaactg taaagtaatt gtgtgttttg agactataaa
7501 tatcccttgg agaaaagcct tgttaacgcg cggtgaccct cgaggtcgac ggtatcgata
7561 agctcgcttc acgagattcc agcaggtcga gggacctaat aacttcgtat agcatacatt
7621 atacgaagtt atattaaggg ttccaagctt aagcggccgc gtggataacc gtattaccgc
7681 catgcattag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
7741 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
7801 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
7861 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
7921 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
7981 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
8041 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
8101 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
8161 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
8221 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct
8281 agcaccaggt atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt
8341 cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga
8401 tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc
8461 ctggcccacc ctcgtgacca ccctgagctg gggcgtgcag tgcttcgccc gctaccccga
8521 ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg
8581 caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg
8641 cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat
8701 cctggggcac aagctggagt acaactactt cagcgacaac gtctatatca ccgccgacaa
8761 gcagaagaac ggcatcaagg ccaacttcaa gatccgccac aacatcgagg acggcggcgt
8821 gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc
8881 cgacaaccac tacctgagca cccagtccaa gctgagcaaa gaccccaacg agaagcgcga
8941 tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct
9001 gtacaagtac tcaatgttgc cttccctggg aggataccca tcacactggc gaccactcga
9061 ggccagcccg ccgtcccccg cgcgatggag cctgggacgg aagcgcagag ccgacggcag
9121 gcgctggagg cccgaagacg ccgaggaggc agagcaccgc ggcgccgagc gcagacccga
9181 gagctttacc actcctgaag gccctaaacc ccgttccaga tgctctgact gggcaagtgc
9241 agttgaagaa gatgaaatga ggaccagagt taacaaagaa atggcaagat ataaaaggaa
9301 actcctcatc aatgactttg gaagagagag aaaatcatca tcaggaagtt ctgattcaaa
9361 ggagtctatg tctactgtgc cggctgacga tcgaggatga agaaggatct tagcttaagg
9421 cccctctccc t
//
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
| P4830/pLVX-IRES-Puro-TGFBR2人源基因质粒 |
| P4831/pLVX-AcGFP1-TEFM人源基因质粒 |
| P0711/pCDNA3.1-HA-SP1人源基因质粒 |
| P0912/pEGFP-CTNNB1人源基因质粒 |
| P0927/pCAGGS-HA-hUBE1L人源基因质粒 |
| P4832/pCAGGS-HA-TRIM25人源基因质粒 |
| P0968/pCDNA3.1-TNFAIP8L3-HA人源基因质粒 |
| P0992/pCMV-N-HA-SIRT2人源基因质粒 |
| P0994/p3×FLAG-CMV-SNX26人源基因质粒 |
| P0778/pEGFP-hsp27 wt FL人源基因质粒 |
| P0790/pCMV-HA-MDM2人源基因质粒 |
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