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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：
NM_001123377.1;DJ1;DJ-1
- 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：pLVX-PARK7-EGFP-IRES-Puro人源基因质粒
别称: NM_001123377.1;DJ1;DJ-1
复制子: pUC
原核抗性: Amp
筛选标记：Puro
克隆菌株: Stbl3
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：Puro
荧光蛋白：
质粒简介：详询
质粒图谱
质粒序列
LOCUS Exported 9413 bp ds-DNA circular SYN 26-SEP-2017
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9413)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Sep 26, 2017 from MiaoLingPlasmid
http://www.miaolingbio.com
FEATURES Location/Qualifiers
source 1..9413
/organism="synthetic DNA construct"
/mol_type="other DNA"
LTR 1..634
/label=3' LTR
/note="3' long terminal repeat (LTR) from HIV-1"
misc_feature 681..806
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1303..1536
/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 2028..2143
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1 (lacking the first T)"
enhancer 2201..2504
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 2505..2708
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
misc_feature 2815..3381
/label=PARK7
/note="Homo sapiens Parkinsonism associated deglycase
(PARK7), transcript variant 2, mRNA
NCBI Reference Sequence: NM_001123377.1
"
CDS 3388..4107
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
misc_feature 4116..4689
/label=IRES
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
CDS 4722..5321
/codon_start=1
/gene="pac from Streptomyces alboniger"
/product="puromycin N-acetyltransferase"
/label=PuroR
/note="confers resistance to puromycin"
/translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
misc_feature 5335..5923
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
CDS complement(5806..5817)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
LTR 6130..6763
/label=3' LTR
/note="3' long terminal repeat (LTR) from HIV-1"
protein_bind 6915..6931
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
promoter complement(6939..6969)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 6984..7005
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(7293..7881)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(8052..8912)
/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(8913..9017)
/gene="bla"
/label=AmpR promoter
polyA_signal 9065..9199
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
ORIGIN
1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccttt aaaagaaaag
2041 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac
2101 aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg
2161 acagcagaga tccagtttat cgataagctt gggagttccg cgttacataa cttacggtaa
2221 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
2281 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
2341 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
2401 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
2461 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc
2521 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca
2581 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta
2641 acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa
2701 gcagagctcg tttagtgaac cgtcagatcg cctggagacg ccatccacgc tgttttgacc
2761 tccatagaag acaccgactc tactagagga tctatttccg gtgaattcgc caccatggct
2821 tccaaaagag ctctggtcat cctggctaaa ggagcagagg aaatggagac ggtcatccct
2881 gtagatgtca tgaggcgagc tgggattaag gtcaccgttg caggcctggc tggaaaagac
2941 ccagtacagt gtagccgtga tgtggtcatt tgtcctgatg ccagccttga agatgcaaaa
3001 aaagagggac catatgatgt ggtggttcta ccaggaggta atctgggcgc acagaattta
3061 tctgagtctg ctgctgtgaa ggagatactg aaggagcagg aaaaccggaa gggcctgata
3121 gccgccatct gtgcaggtcc tactgctctg ttggctcatg aaataggttt tggaagtaaa
3181 gttacaacac accctcttgc taaagacaaa atgatgaatg gaggtcatta cacctactct
3241 gagaatcgtg tggaaaaaga cggcctgatt cttacaagcc gggggcctgg gaccagcttc
3301 gagtttgcgc ttgcaattgt tgaagccctg aatggcaagg aggtggcggc tcaagtgaag
3361 gctccacttg ttcttaaaga ctctagaatg gtgagcaagg gcgaggagct gttcaccggg
3421 gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc
3481 ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc
3541 ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc
3601 ttcagccgct accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa
3661 ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc
3721 gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc
3781 aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc
3841 tatatcatgg ccgacaagca gaagaacggc atcaaggtga acttcaagat ccgccacaac
3901 atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac
3961 ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccgccct gagcaaagac
4021 cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact
4081 ctcggcatgg acgagctgta caagtaagga tcccgcccct ctccctcccc cccccctaac
4141 gttactggcc gaagccgctt ggaataaggc cggtgtgcgt ttgtctatat gttattttcc
4201 accatattgc cgtcttttgg caatgtgagg gcccggaaac ctggccctgt cttcttgacg
4261 agcattccta ggggtctttc ccctctcgcc aaaggaatgc aaggtctgtt gaatgtcgtg
4321 aaggaagcag ttcctctgga agcttcttga agacaaacaa cgtctgtagc gaccctttgc
4381 aggcagcgga accccccacc tggcgacagg tgcctctgcg gccaaaagcc acgtgtataa
4441 gatacacctg caaaggcggc acaaccccag tgccacgttg tgagttggat agttgtggaa
4501 agagtcaaat ggctctcctc aagcgtattc aacaaggggc tgaaggatgc ccagaaggta
4561 ccccattgta tgggatctga tctggggcct cggtgcacat gctttacatg tgtttagtcg
4621 aggttaaaaa aacgtctagg ccccccgaac cacggggacg tggttttcct ttgaaaaaca
4681 cgatgataag cttgccacaa cccacaagga gacgaccttc catgaccgag tacaagccca
4741 cggtgcgcct cgccacccgc gacgacgtcc cccgggccgt acgcaccctc gccgccgcgt
4801 tcgccgacta ccccgccacg cgccacaccg tcgacccgga ccgccacatc gagcgggtca
4861 ccgagctgca agaactcttc ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg
4921 cggacgacgg cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg
4981 tgttcgccga gatcggcccg cgcatggccg agttgagcgg ttcccggctg gccgcgcagc
5041 aacagatgga aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca
5101 ccgtcggcgt ctcgcccgac caccagggca agggtctggg cagcgccgtc gtgctccccg
5161 gagtggaggc ggccgagcgc gccggggtgc ccgccttcct ggagacctcc gcgccccgca
5221 acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag
5281 gaccgcgcac ctggtgcatg acccgcaagc ccggtgccta gacgcgtctg gaacaatcaa
5341 cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt
5401 acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct
5461 ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc
5521 gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg
5581 ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc
5641 acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc
5701 actgacaatt ccgtggtgtt gtcggggaag ctgacgtcct ttccatggct gctcgcctgt
5761 gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca
5821 gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt
5881 cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcctggaat taattctgca
5941 gtcgagacct agaaaaacat ggagcaatca caagtagcaa tacagcagct accaatgctg
6001 attgtgcctg gctagaagca caagaggagg aggaggtggg ttttccagtc acacctcagg
6061 tacctttaag accaatgact tacaaggcag ctgtagatct tagccacttt ttaaaagaaa
6121 agaggggact ggaagggcta attcactccc aacgaagaca agatatcctt gatctgtgga
6181 tctaccacac acaaggctac ttccctgatt agcagaacta cacaccaggg ccaggggtca
6241 gatatccact gacctttgga tggtgctaca agctagtacc agttgagcca gataaggtag
6301 aagaggccaa taaaggagag aacaccagct tgttacaccc tgtgagcctg catgggatgg
6361 atgacccgga gagagaagtg ttagagtgga ggtttgacag ccgcctagca tttcatcacg
6421 tggcccgaga gctgcatccg gagtacttca agaactgctg atatcgagct tgctacaagg
6481 gactttccgc tggggacttt ccagggaggc gtggcctggg cgggactggg gagtggcgag
6541 ccctcagatc ctgcatataa gcagctgctt tttgcctgta ctgggtctct ctggttagac
6601 cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa
6661 agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag
6721 agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtagtag ttcatgtcat
6781 cttattattc agtatttata acttgcaaag aaatgaatat cagagagtga gaggccttga
6841 cattgctagc gtttaccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg
6901 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
6961 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
7021 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
7081 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
7141 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
7201 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
7261 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
7321 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
7381 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
7441 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
7501 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
7561 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
7621 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
7681 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
7741 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
7801 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
7861 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
7921 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
7981 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
8041 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
8101 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
8161 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
8221 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
8281 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
8341 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
8401 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
8461 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
8521 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
8581 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
8641 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
8701 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
8761 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
8821 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
8881 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
8941 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
9001 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga cggatcggga
9061 gatcaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
9121 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
9181 atcttatcat gtctggatca actggataac tcaagctaac caaaatcatc ccaaacttcc
9241 caccccatac cctattacca ctgccaatta cctgtggttt catttactct aaacctgtga
9301 ttcctctgaa ttattttcat tttaaagaaa ttgtatttgt taaatatgta ctacaaactt
9361 agtagttttt aaagaaattg tatttgttaa atatgtacta caaacttagt agt
//
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：pLVX-PARK7-EGFP-IRES-Puro人源基因质粒
别称: NM_001123377.1;DJ1;DJ-1
复制子: pUC
原核抗性: Amp
筛选标记：Puro
克隆菌株: Stbl3
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：Puro
荧光蛋白：
质粒简介：详询
质粒图谱
质粒序列
LOCUS Exported 9413 bp ds-DNA circular SYN 26-SEP-2017
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9413)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Sep 26, 2017 from MiaoLingPlasmid
http://www.miaolingbio.com
FEATURES Location/Qualifiers
source 1..9413
/organism="synthetic DNA construct"
/mol_type="other DNA"
LTR 1..634
/label=3' LTR
/note="3' long terminal repeat (LTR) from HIV-1"
misc_feature 681..806
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1303..1536
/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 2028..2143
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1 (lacking the first T)"
enhancer 2201..2504
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 2505..2708
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
misc_feature 2815..3381
/label=PARK7
/note="Homo sapiens Parkinsonism associated deglycase
(PARK7), transcript variant 2, mRNA
NCBI Reference Sequence: NM_001123377.1
"
CDS 3388..4107
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
misc_feature 4116..4689
/label=IRES
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
CDS 4722..5321
/codon_start=1
/gene="pac from Streptomyces alboniger"
/product="puromycin N-acetyltransferase"
/label=PuroR
/note="confers resistance to puromycin"
/translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
misc_feature 5335..5923
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
CDS complement(5806..5817)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
LTR 6130..6763
/label=3' LTR
/note="3' long terminal repeat (LTR) from HIV-1"
protein_bind 6915..6931
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
promoter complement(6939..6969)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 6984..7005
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(7293..7881)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(8052..8912)
/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(8913..9017)
/gene="bla"
/label=AmpR promoter
polyA_signal 9065..9199
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
ORIGIN
1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccttt aaaagaaaag
2041 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac
2101 aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg
2161 acagcagaga tccagtttat cgataagctt gggagttccg cgttacataa cttacggtaa
2221 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
2281 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
2341 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
2401 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
2461 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc
2521 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca
2581 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta
2641 acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa
2701 gcagagctcg tttagtgaac cgtcagatcg cctggagacg ccatccacgc tgttttgacc
2761 tccatagaag acaccgactc tactagagga tctatttccg gtgaattcgc caccatggct
2821 tccaaaagag ctctggtcat cctggctaaa ggagcagagg aaatggagac ggtcatccct
2881 gtagatgtca tgaggcgagc tgggattaag gtcaccgttg caggcctggc tggaaaagac
2941 ccagtacagt gtagccgtga tgtggtcatt tgtcctgatg ccagccttga agatgcaaaa
3001 aaagagggac catatgatgt ggtggttcta ccaggaggta atctgggcgc acagaattta
3061 tctgagtctg ctgctgtgaa ggagatactg aaggagcagg aaaaccggaa gggcctgata
3121 gccgccatct gtgcaggtcc tactgctctg ttggctcatg aaataggttt tggaagtaaa
3181 gttacaacac accctcttgc taaagacaaa atgatgaatg gaggtcatta cacctactct
3241 gagaatcgtg tggaaaaaga cggcctgatt cttacaagcc gggggcctgg gaccagcttc
3301 gagtttgcgc ttgcaattgt tgaagccctg aatggcaagg aggtggcggc tcaagtgaag
3361 gctccacttg ttcttaaaga ctctagaatg gtgagcaagg gcgaggagct gttcaccggg
3421 gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc
3481 ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc
3541 ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc
3601 ttcagccgct accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa
3661 ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc
3721 gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc
3781 aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc
3841 tatatcatgg ccgacaagca gaagaacggc atcaaggtga acttcaagat ccgccacaac
3901 atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac
3961 ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccgccct gagcaaagac
4021 cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact
4081 ctcggcatgg acgagctgta caagtaagga tcccgcccct ctccctcccc cccccctaac
4141 gttactggcc gaagccgctt ggaataaggc cggtgtgcgt ttgtctatat gttattttcc
4201 accatattgc cgtcttttgg caatgtgagg gcccggaaac ctggccctgt cttcttgacg
4261 agcattccta ggggtctttc ccctctcgcc aaaggaatgc aaggtctgtt gaatgtcgtg
4321 aaggaagcag ttcctctgga agcttcttga agacaaacaa cgtctgtagc gaccctttgc
4381 aggcagcgga accccccacc tggcgacagg tgcctctgcg gccaaaagcc acgtgtataa
4441 gatacacctg caaaggcggc acaaccccag tgccacgttg tgagttggat agttgtggaa
4501 agagtcaaat ggctctcctc aagcgtattc aacaaggggc tgaaggatgc ccagaaggta
4561 ccccattgta tgggatctga tctggggcct cggtgcacat gctttacatg tgtttagtcg
4621 aggttaaaaa aacgtctagg ccccccgaac cacggggacg tggttttcct ttgaaaaaca
4681 cgatgataag cttgccacaa cccacaagga gacgaccttc catgaccgag tacaagccca
4741 cggtgcgcct cgccacccgc gacgacgtcc cccgggccgt acgcaccctc gccgccgcgt
4801 tcgccgacta ccccgccacg cgccacaccg tcgacccgga ccgccacatc gagcgggtca
4861 ccgagctgca agaactcttc ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg
4921 cggacgacgg cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg
4981 tgttcgccga gatcggcccg cgcatggccg agttgagcgg ttcccggctg gccgcgcagc
5041 aacagatgga aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca
5101 ccgtcggcgt ctcgcccgac caccagggca agggtctggg cagcgccgtc gtgctccccg
5161 gagtggaggc ggccgagcgc gccggggtgc ccgccttcct ggagacctcc gcgccccgca
5221 acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag
5281 gaccgcgcac ctggtgcatg acccgcaagc ccggtgccta gacgcgtctg gaacaatcaa
5341 cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt
5401 acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct
5461 ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc
5521 gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg
5581 ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc
5641 acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc
5701 actgacaatt ccgtggtgtt gtcggggaag ctgacgtcct ttccatggct gctcgcctgt
5761 gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca
5821 gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt
5881 cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcctggaat taattctgca
5941 gtcgagacct agaaaaacat ggagcaatca caagtagcaa tacagcagct accaatgctg
6001 attgtgcctg gctagaagca caagaggagg aggaggtggg ttttccagtc acacctcagg
6061 tacctttaag accaatgact tacaaggcag ctgtagatct tagccacttt ttaaaagaaa
6121 agaggggact ggaagggcta attcactccc aacgaagaca agatatcctt gatctgtgga
6181 tctaccacac acaaggctac ttccctgatt agcagaacta cacaccaggg ccaggggtca
6241 gatatccact gacctttgga tggtgctaca agctagtacc agttgagcca gataaggtag
6301 aagaggccaa taaaggagag aacaccagct tgttacaccc tgtgagcctg catgggatgg
6361 atgacccgga gagagaagtg ttagagtgga ggtttgacag ccgcctagca tttcatcacg
6421 tggcccgaga gctgcatccg gagtacttca agaactgctg atatcgagct tgctacaagg
6481 gactttccgc tggggacttt ccagggaggc gtggcctggg cgggactggg gagtggcgag
6541 ccctcagatc ctgcatataa gcagctgctt tttgcctgta ctgggtctct ctggttagac
6601 cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa
6661 agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag
6721 agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtagtag ttcatgtcat
6781 cttattattc agtatttata acttgcaaag aaatgaatat cagagagtga gaggccttga
6841 cattgctagc gtttaccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg
6901 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
6961 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
7021 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
7081 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
7141 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
7201 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
7261 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
7321 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
7381 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
7441 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
7501 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
7561 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
7621 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
7681 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
7741 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
7801 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
7861 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
7921 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
7981 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
8041 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
8101 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
8161 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
8221 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
8281 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
8341 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
8401 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
8461 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
8521 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
8581 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
8641 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
8701 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
8761 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
8821 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
8881 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
8941 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
9001 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga cggatcggga
9061 gatcaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
9121 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
9181 atcttatcat gtctggatca actggataac tcaagctaac caaaatcatc ccaaacttcc
9241 caccccatac cctattacca ctgccaatta cctgtggttt catttactct aaacctgtga
9301 ttcctctgaa ttattttcat tttaaagaaa ttgtatttgt taaatatgta ctacaaactt
9361 agtagttttt aaagaaattg tatttgttaa atatgtacta caaacttagt agt
//
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
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文献和实验相关实验
-Puro 病毒质粒 Amp pLVX-shRNA 病毒质粒 Amp pLVX-IRES-Zsgreen1 病毒质粒 Amp pLVX-AcGFP-N1 病毒质粒
PYM30 ECGP PYM33 EBFP PYM39 EYFP pLVX-ShRNA1 pLVX-puro PLVX-ires-zsgreen1 PLVX-acgfp-n1 PLVX-dsred-monomer-n1
细胞株。此外， 载体中的pUC origin 能保证该载体在大肠杆菌中的复制， 而位于此表达盒上游的细菌启动子能驱动kanamycin抗性基因在大肠杆菌中的表达。 用途: 该表达载体EGFP上游有Nde1、Eco47111和Age1克隆位点，将外源基因扦入这些位点，将合成外源基因和EGFP的融合基因。 借此可确定外源基因在细胞内的表达和/或组织中的定位。 亦可用于检测克隆的启动子活性(取代CMV启动子,Acet1-Nhe1)。 3、pEGFT-Actin, 增强型绿色荧光蛋白/人肌
pLVX-PARK7-EGFP-IRES-Puro人源基因质粒
￥100 - 1000
