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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_001108867.2
- 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-prmt5-r-IRES-Puro大鼠基因质粒
别称: NM_001108867.2
启动子：CMV
复制子: pUC
终止子: WPRE
质粒分类: 基因文库质粒；大鼠基因质粒；大鼠真核质粒
质粒大小：10741bp
质粒标签：N-HA
原核抗性: Amp
筛选标记：Puro
克隆菌株: Stbl3
培养条件: 37度
表达宿主: 293T等哺乳细胞
质粒属性
载体宿主：哺乳细胞,慢病毒
载体用途：蛋白表达
基因种属：大鼠
基因类型：ORF
原核抗性：Amp
真核抗性：Puro
荧光蛋白：
质粒简介
pLVX-prmt5-r-IRES-Puro质粒是在pLVX-IRES-Puro背景载体上通过EcoR I 和NotI 酶切位点克隆进入大鼠prmt5基因，CMV启动子驱动prmt5大鼠源基因表达。pLVX-prmt5-r-IRES-Puro质粒由武汉淼灵生物科技有限公司构建并分享至淼灵质粒平台。
Rattus norvegicus lysine demethylase 1A (Kdm1a), mRNA.NCBI Reference Sequence: NM_001130098.1.
质粒图谱
质粒序列
LOCUS Exported 10741 bp ds-DNA circular SYN 15-JUN-2017
DEFINITION synthetic circular DNA
FEATURES Location/Qualifiers
source 1..10741
/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..2144
/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 2803..5435
/label=Kdm1aT-r
misc_feature 5444..6017
/label=IRES
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
CDS 6050..6649
/codon_start=1
/gene="pac from Streptomyces alboniger"
/product="puromycin N-acetyltransferase"
/label=Puro
/note="confers resistance to puromycin"
/translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
misc_feature 6663..7251
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
CDS complement(7134..7145)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
LTR 7458..8091
/label=3' LTR
/note="3' long terminal repeat (LTR) from HIV-1"
primer_bind complement(8219..8235)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 8243..8259
/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(8267..8297)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 8312..8333
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(8621..9209)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(9380..10240)
/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(10241..10345)
/gene="bla"
/label=AmpR promoter
polyA_signal 10393..10527
/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 gtgaattcat gttgtctggg
2821 aagaaggcgg cggcggcggc agcggcggcg gcggcagcag cggctgggac agaggccggg
2881 cccggggcgg cgggcggcgc cgagaacggc tctgaggtgg ccgcgccgcc cgcgggcctg
2941 gcgggccccg ccgacattgc tacgggggcg gcgggcgagc gcactccccg aaagaaggag
3001 cctccgcggg cctcgccgcc cggaggcctg gccgagccgc ctgggtccgc cgggcctcag
3061 gcggggccca cagccgggcc cggctccgct acacccatgg agacaggaat agccgagacc
3121 ccggagggcc gacggaccag ccgacgcaag cgggcgaagg tagaatacag agaaatggat
3181 gaaagcttgg ccaacctctc agaagatgaa tattattcag aagaagaaag aaatgccaaa
3241 gcagagaagg agaagaagct tcccccgcca cctcctcaag ccccacctga ggaggaaaat
3301 gaaagtgagc ccgaggagcc gtctgggcaa gcaggaggac ttcaagacga cagttctgga
3361 gggtatggag acggccaagc atcaggtgtg gagggtgcag ctttccaaag cagacttccc
3421 catgaccgaa tgacctctca ggaagcagcg tgtttcccag acatcatcag tgggcctcag
3481 cagacacaga aggtttttct gttcatcagg aatcgcacat tgcagttatg gttggacaac
3541 ccaaagattc agctgacatt tgaagctacg ctccagcagt tagaagcacc ttataacagt
3601 gacactgtac ttgtccaccg agttcatagt tacttagagc gccacggtct tatcaacttc
3661 ggcatctaca agaggataaa acctttacca attaaaaaga caggaaaggt gattattata
3721 ggctcaggtg tttctggctt ggcagcagct cgacaactac agagttttgg gatggatgtc
3781 acacttctgg aagccaggga tcgcgtaggt ggacgggtcg ctacatttcg aaaaggaaac
3841 tatgtagctg atcttggagc catggtagta acaggtcttg gaggaaatcc catggctgtc
3901 gtcagcaaac aagtaaatat ggaactggcc aagatcaagc aaaaatgccc actttatgaa
3961 gccaatggac aagctgttcc aaaagagaaa gatgaaatgg tagaacaaga atttaaccgg
4021 ttgcttgaag ccacctctta ccttagtcat cagttagact tcaacgtcct caataataaa
4081 cctgtgtccc ttggccaggc gttggaggtt gtcattcagc tacaagaaaa gcacgtcaaa
4141 gatgagcaga ttgaacactg gaagaagata gtgaaaactc aggaggaatt gaaagagctt
4201 ctcaataaga tggtaaactt gaaggagaaa attaaagagc tccaccagca atacaaagaa
4261 gcttcagaag tgaagccgcc cagagatatt acagctgagt tcctagtgaa aagcaagcac
4321 agggacctga ctgccctctg caaggaatat gatgaattag ctgaaacaca aggaaagcta
4381 gaagaaaaac ttcaagaatt ggaagccaat cccccaagtg atgtatatct ctcatcaaga
4441 gacagacaaa tacttgactg gcattttgca aatcttgaat ttgccaacgc cacacctctc
4501 tctaccctct ctcttaaaca ttgggatcag gatgatgact tcgagtttac tggcagccac
4561 ctgacagtga ggaatggcta ctcgtgtgtg cctgtggctt tagctgaagg tttggacatt
4621 aaactgaaca cagcagtgcg gcaggttcgc tacacagcct caggatgtga agtgattgct
4681 gtgaacacac gctccacaag tcagaccttc atttataagt gtgacgcagt gctctgtacc
4741 cttcctttgg gagtgctgaa gcagcagcca ccagctgttc agtttgtgcc gcctcttcct
4801 gagtggaaaa catccgcagt ccaaaggatg ggatttggca accttaacaa ggtggtgcta
4861 tgttttgacc gtgtgttctg ggatccaagt gtcaatttgt ttgggcacgt tggcagtaca
4921 actgctagca ggggcgagct cttcctcttc tggaacttat ataaagctcc aatactcttg
4981 gctctggtag caggcgaagc tgctggcatt atggagaaca ttagtgatga tgtgattgtc
5041 ggccggtgcc tggccattct caaagggatt tttggcagca gtgcagtacc acagcccaag
5101 gaaactgtgg tgtctcgttg gcgtgctgat ccctgggccc ggggctccta ttcttatgtg
5161 gctgcaggat cttctggaaa tgactatgat ttaatggctc agccaatcac tcctggtccc
5221 tcaattccag gtgccccaca gccaattcca agactcttct ttgctggaga acatacaatc
5281 cgcaactacc cagctacggt ccatggtgct ctgctgagtg ggcttcgaga agcaggaagg
5341 attgccgacc agtttttggg agccatgtac acgttgcctc gtcaggccac accgggtgtc
5401 cctgcacagc agtccccaag tatgtgagcg gccgcggatc ccgcccctct ccctcccccc
5461 cccctaacgt tactggccga agccgcttgg aataaggccg gtgtgcgttt gtctatatgt
5521 tattttccac catattgccg tcttttggca atgtgagggc ccggaaacct ggccctgtct
5581 tcttgacgag cattcctagg ggtctttccc ctctcgccaa aggaatgcaa ggtctgttga
5641 atgtcgtgaa ggaagcagtt cctctggaag cttcttgaag acaaacaacg tctgtagcga
5701 ccctttgcag gcagcggaac cccccacctg gcgacaggtg cctctgcggc caaaagccac
5761 gtgtataaga tacacctgca aaggcggcac aaccccagtg ccacgttgtg agttggatag
5821 ttgtggaaag agtcaaatgg ctctcctcaa gcgtattcaa caaggggctg aaggatgccc
5881 agaaggtacc ccattgtatg ggatctgatc tggggcctcg gtgcacatgc tttacatgtg
5941 tttagtcgag gttaaaaaaa cgtctaggcc ccccgaacca cggggacgtg gttttccttt
6001 gaaaaacacg atgataagct tgccacaacc cacaaggaga cgaccttcca tgaccgagta
6061 caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc cgggccgtac gcaccctcgc
6121 cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc gacccggacc gccacatcga
6181 gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt
6241 gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc
6301 gggggcggtg ttcgccgaga tcggcccgcg catggccgag ttgagcggtt cccggctggc
6361 cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt
6421 cctggccacc gtcggcgtct cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt
6481 gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg agacctccgc
6541 gccccgcaac ctccccttct acgagcggct cggcttcacc gtcaccgccg acgtcgaggt
6601 gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctaga cgcgtctgga
6661 acaatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg
6721 ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc
6781 gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt
6841 tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca
6901 ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc
6961 ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc
7021 tgttgggcac tgacaattcc gtggtgttgt cggggaagct gacgtccttt ccatggctgc
7081 tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc
7141 tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc
7201 ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg cctggaatta
7261 attctgcagt cgagacctag aaaaacatgg agcaatcaca agtagcaata cagcagctac
7321 caatgctgat tgtgcctggc tagaagcaca agaggaggag gaggtgggtt ttccagtcac
7381 acctcaggta cctttaagac caatgactta caaggcagct gtagatctta gccacttttt
7441 aaaagaaaag aggggactgg aagggctaat tcactcccaa cgaagacaag atatccttga
7501 tctgtggatc taccacacac aaggctactt ccctgattag cagaactaca caccagggcc
7561 aggggtcaga tatccactga cctttggatg gtgctacaag ctagtaccag ttgagccaga
7621 taaggtagaa gaggccaata aaggagagaa caccagcttg ttacaccctg tgagcctgca
7681 tgggatggat gacccggaga gagaagtgtt agagtggagg tttgacagcc gcctagcatt
7741 tcatcacgtg gcccgagagc tgcatccgga gtacttcaag aactgctgat atcgagcttg
7801 ctacaaggga ctttccgctg gggactttcc agggaggcgt ggcctgggcg ggactgggga
7861 gtggcgagcc ctcagatcct gcatataagc agctgctttt tgcctgtact gggtctctct
7921 ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc
7981 ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg
8041 gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc agtagtagtt
8101 catgtcatct tattattcag tatttataac ttgcaaagaa atgaatatca gagagtgaga
8161 ggccttgaca ttgctagcgt ttaccgtcga cctctagcta gagcttggcg taatcatggt
8221 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
8281 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
8341 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
8401 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
8461 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
8521 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
8581 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
8641 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
8701 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
8761 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
8821 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
8881 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
8941 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
9001 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
9061 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
9121 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
9181 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
9241 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
9301 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
9361 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
9421 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
9481 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
9541 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
9601 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
9661 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
9721 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
9781 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
9841 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
9901 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
9961 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
10021 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
10081 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
10141 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
10201 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
10261 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
10321 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg
10381 gatcgggaga tcaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
10441 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
10501 atcaatgtat cttatcatgt ctggatcaac tggataactc aagctaacca aaatcatccc
10561 aaacttccca ccccataccc tattaccact gccaattacc tgtggtttca tttactctaa
10621 acctgtgatt cctctgaatt attttcattt taaagaaatt gtatttgtta aatatgtact
10681 acaaacttag tagtttttaa agaaattgta tttgttaaat atgtactaca aacttagtag
10741 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：pLVX-prmt5-r-IRES-Puro大鼠基因质粒
别称: NM_001108867.2
启动子：CMV
复制子: pUC
终止子: WPRE
质粒分类: 基因文库质粒；大鼠基因质粒；大鼠真核质粒
质粒大小：10741bp
质粒标签：N-HA
原核抗性: Amp
筛选标记：Puro
克隆菌株: Stbl3
培养条件: 37度
表达宿主: 293T等哺乳细胞
质粒属性
载体宿主：哺乳细胞,慢病毒
载体用途：蛋白表达
基因种属：大鼠
基因类型：ORF
原核抗性：Amp
真核抗性：Puro
荧光蛋白：
质粒简介
pLVX-prmt5-r-IRES-Puro质粒是在pLVX-IRES-Puro背景载体上通过EcoR I 和NotI 酶切位点克隆进入大鼠prmt5基因，CMV启动子驱动prmt5大鼠源基因表达。pLVX-prmt5-r-IRES-Puro质粒由武汉淼灵生物科技有限公司构建并分享至淼灵质粒平台。
Rattus norvegicus lysine demethylase 1A (Kdm1a), mRNA.NCBI Reference Sequence: NM_001130098.1.
质粒图谱
质粒序列
LOCUS Exported 10741 bp ds-DNA circular SYN 15-JUN-2017
DEFINITION synthetic circular DNA
FEATURES Location/Qualifiers
source 1..10741
/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..2144
/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 2803..5435
/label=Kdm1aT-r
misc_feature 5444..6017
/label=IRES
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
CDS 6050..6649
/codon_start=1
/gene="pac from Streptomyces alboniger"
/product="puromycin N-acetyltransferase"
/label=Puro
/note="confers resistance to puromycin"
/translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
misc_feature 6663..7251
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
CDS complement(7134..7145)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
LTR 7458..8091
/label=3' LTR
/note="3' long terminal repeat (LTR) from HIV-1"
primer_bind complement(8219..8235)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 8243..8259
/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(8267..8297)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 8312..8333
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(8621..9209)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(9380..10240)
/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(10241..10345)
/gene="bla"
/label=AmpR promoter
polyA_signal 10393..10527
/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 gtgaattcat gttgtctggg
2821 aagaaggcgg cggcggcggc agcggcggcg gcggcagcag cggctgggac agaggccggg
2881 cccggggcgg cgggcggcgc cgagaacggc tctgaggtgg ccgcgccgcc cgcgggcctg
2941 gcgggccccg ccgacattgc tacgggggcg gcgggcgagc gcactccccg aaagaaggag
3001 cctccgcggg cctcgccgcc cggaggcctg gccgagccgc ctgggtccgc cgggcctcag
3061 gcggggccca cagccgggcc cggctccgct acacccatgg agacaggaat agccgagacc
3121 ccggagggcc gacggaccag ccgacgcaag cgggcgaagg tagaatacag agaaatggat
3181 gaaagcttgg ccaacctctc agaagatgaa tattattcag aagaagaaag aaatgccaaa
3241 gcagagaagg agaagaagct tcccccgcca cctcctcaag ccccacctga ggaggaaaat
3301 gaaagtgagc ccgaggagcc gtctgggcaa gcaggaggac ttcaagacga cagttctgga
3361 gggtatggag acggccaagc atcaggtgtg gagggtgcag ctttccaaag cagacttccc
3421 catgaccgaa tgacctctca ggaagcagcg tgtttcccag acatcatcag tgggcctcag
3481 cagacacaga aggtttttct gttcatcagg aatcgcacat tgcagttatg gttggacaac
3541 ccaaagattc agctgacatt tgaagctacg ctccagcagt tagaagcacc ttataacagt
3601 gacactgtac ttgtccaccg agttcatagt tacttagagc gccacggtct tatcaacttc
3661 ggcatctaca agaggataaa acctttacca attaaaaaga caggaaaggt gattattata
3721 ggctcaggtg tttctggctt ggcagcagct cgacaactac agagttttgg gatggatgtc
3781 acacttctgg aagccaggga tcgcgtaggt ggacgggtcg ctacatttcg aaaaggaaac
3841 tatgtagctg atcttggagc catggtagta acaggtcttg gaggaaatcc catggctgtc
3901 gtcagcaaac aagtaaatat ggaactggcc aagatcaagc aaaaatgccc actttatgaa
3961 gccaatggac aagctgttcc aaaagagaaa gatgaaatgg tagaacaaga atttaaccgg
4021 ttgcttgaag ccacctctta ccttagtcat cagttagact tcaacgtcct caataataaa
4081 cctgtgtccc ttggccaggc gttggaggtt gtcattcagc tacaagaaaa gcacgtcaaa
4141 gatgagcaga ttgaacactg gaagaagata gtgaaaactc aggaggaatt gaaagagctt
4201 ctcaataaga tggtaaactt gaaggagaaa attaaagagc tccaccagca atacaaagaa
4261 gcttcagaag tgaagccgcc cagagatatt acagctgagt tcctagtgaa aagcaagcac
4321 agggacctga ctgccctctg caaggaatat gatgaattag ctgaaacaca aggaaagcta
4381 gaagaaaaac ttcaagaatt ggaagccaat cccccaagtg atgtatatct ctcatcaaga
4441 gacagacaaa tacttgactg gcattttgca aatcttgaat ttgccaacgc cacacctctc
4501 tctaccctct ctcttaaaca ttgggatcag gatgatgact tcgagtttac tggcagccac
4561 ctgacagtga ggaatggcta ctcgtgtgtg cctgtggctt tagctgaagg tttggacatt
4621 aaactgaaca cagcagtgcg gcaggttcgc tacacagcct caggatgtga agtgattgct
4681 gtgaacacac gctccacaag tcagaccttc atttataagt gtgacgcagt gctctgtacc
4741 cttcctttgg gagtgctgaa gcagcagcca ccagctgttc agtttgtgcc gcctcttcct
4801 gagtggaaaa catccgcagt ccaaaggatg ggatttggca accttaacaa ggtggtgcta
4861 tgttttgacc gtgtgttctg ggatccaagt gtcaatttgt ttgggcacgt tggcagtaca
4921 actgctagca ggggcgagct cttcctcttc tggaacttat ataaagctcc aatactcttg
4981 gctctggtag caggcgaagc tgctggcatt atggagaaca ttagtgatga tgtgattgtc
5041 ggccggtgcc tggccattct caaagggatt tttggcagca gtgcagtacc acagcccaag
5101 gaaactgtgg tgtctcgttg gcgtgctgat ccctgggccc ggggctccta ttcttatgtg
5161 gctgcaggat cttctggaaa tgactatgat ttaatggctc agccaatcac tcctggtccc
5221 tcaattccag gtgccccaca gccaattcca agactcttct ttgctggaga acatacaatc
5281 cgcaactacc cagctacggt ccatggtgct ctgctgagtg ggcttcgaga agcaggaagg
5341 attgccgacc agtttttggg agccatgtac acgttgcctc gtcaggccac accgggtgtc
5401 cctgcacagc agtccccaag tatgtgagcg gccgcggatc ccgcccctct ccctcccccc
5461 cccctaacgt tactggccga agccgcttgg aataaggccg gtgtgcgttt gtctatatgt
5521 tattttccac catattgccg tcttttggca atgtgagggc ccggaaacct ggccctgtct
5581 tcttgacgag cattcctagg ggtctttccc ctctcgccaa aggaatgcaa ggtctgttga
5641 atgtcgtgaa ggaagcagtt cctctggaag cttcttgaag acaaacaacg tctgtagcga
5701 ccctttgcag gcagcggaac cccccacctg gcgacaggtg cctctgcggc caaaagccac
5761 gtgtataaga tacacctgca aaggcggcac aaccccagtg ccacgttgtg agttggatag
5821 ttgtggaaag agtcaaatgg ctctcctcaa gcgtattcaa caaggggctg aaggatgccc
5881 agaaggtacc ccattgtatg ggatctgatc tggggcctcg gtgcacatgc tttacatgtg
5941 tttagtcgag gttaaaaaaa cgtctaggcc ccccgaacca cggggacgtg gttttccttt
6001 gaaaaacacg atgataagct tgccacaacc cacaaggaga cgaccttcca tgaccgagta
6061 caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc cgggccgtac gcaccctcgc
6121 cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc gacccggacc gccacatcga
6181 gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt
6241 gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc
6301 gggggcggtg ttcgccgaga tcggcccgcg catggccgag ttgagcggtt cccggctggc
6361 cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt
6421 cctggccacc gtcggcgtct cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt
6481 gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg agacctccgc
6541 gccccgcaac ctccccttct acgagcggct cggcttcacc gtcaccgccg acgtcgaggt
6601 gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctaga cgcgtctgga
6661 acaatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg
6721 ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc
6781 gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt
6841 tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca
6901 ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc
6961 ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc
7021 tgttgggcac tgacaattcc gtggtgttgt cggggaagct gacgtccttt ccatggctgc
7081 tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc
7141 tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc
7201 ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg cctggaatta
7261 attctgcagt cgagacctag aaaaacatgg agcaatcaca agtagcaata cagcagctac
7321 caatgctgat tgtgcctggc tagaagcaca agaggaggag gaggtgggtt ttccagtcac
7381 acctcaggta cctttaagac caatgactta caaggcagct gtagatctta gccacttttt
7441 aaaagaaaag aggggactgg aagggctaat tcactcccaa cgaagacaag atatccttga
7501 tctgtggatc taccacacac aaggctactt ccctgattag cagaactaca caccagggcc
7561 aggggtcaga tatccactga cctttggatg gtgctacaag ctagtaccag ttgagccaga
7621 taaggtagaa gaggccaata aaggagagaa caccagcttg ttacaccctg tgagcctgca
7681 tgggatggat gacccggaga gagaagtgtt agagtggagg tttgacagcc gcctagcatt
7741 tcatcacgtg gcccgagagc tgcatccgga gtacttcaag aactgctgat atcgagcttg
7801 ctacaaggga ctttccgctg gggactttcc agggaggcgt ggcctgggcg ggactgggga
7861 gtggcgagcc ctcagatcct gcatataagc agctgctttt tgcctgtact gggtctctct
7921 ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc
7981 ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg
8041 gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc agtagtagtt
8101 catgtcatct tattattcag tatttataac ttgcaaagaa atgaatatca gagagtgaga
8161 ggccttgaca ttgctagcgt ttaccgtcga cctctagcta gagcttggcg taatcatggt
8221 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
8281 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
8341 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
8401 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
8461 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
8521 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
8581 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
8641 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
8701 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
8761 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
8821 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
8881 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
8941 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
9001 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
9061 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
9121 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
9181 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
9241 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
9301 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
9361 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
9421 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
9481 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
9541 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
9601 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
9661 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
9721 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
9781 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
9841 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
9901 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
9961 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
10021 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
10081 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
10141 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
10201 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
10261 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
10321 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg
10381 gatcgggaga tcaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
10441 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
10501 atcaatgtat cttatcatgt ctggatcaac tggataactc aagctaacca aaatcatccc
10561 aaacttccca ccccataccc tattaccact gccaattacc tgtggtttca tttactctaa
10621 acctgtgatt cctctgaatt attttcattt taaagaaatt gtatttgtta aatatgtact
10681 acaaacttag tagtttttaa agaaattgta tttgttaaat atgtactaca aacttagtag
10741 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
| P3627/pCMV-SPORT6-OTULINL人源基因质粒 |
| P3628/pCMV-SPORT6-PDE9A（2同义突变）人源基因质粒 |
| P3629/pCMV-SPORT6-PEX11A（1同义突变）人源基因质粒 |
| P3630/pCMV-SPORT6-CXorf40B人源基因质粒 |
| P3631/pCMV-SPORT6-TMCO6人源基因质粒 |
| P3632/pCMV-SPORT6-SYF2人源基因质粒 |
| P3633/pCMV-SPORT6-PTMS人源基因质粒 |
| P3634/pCMV-SPORT6-STX8（2同义突变）人源基因质粒 |
| P3635/pCMV-SPORT6-APOC1人源基因质粒 |
| P3636/pCMV-SPORT6-GNA15人源基因质粒 |
| P3637/pCMV-SPORT6-RPL18人源基因质粒 |
| P3638/pCMV-SPORT6-NEIL1（1点突变）人源基因质粒 |
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pLVX-prmt5-r-IRES-Puro大鼠基因质粒
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