pLV3-EF1a-CD47-scFv-3×FLAG-PGK

pLV3-EF1a-CD47-scFv-3×FLAG-PGK-CopGFP-Puro plasmid

PVT86354

pLV3-EF1a-CD47-scFv-3×FLAG-PGK-CopGFP-Puro plasmid Information

Plasmid name: pLV3-EF1a-CD47-scFv-3×FLAG-PGK-CopGFP-Puro
Alias:
Gene lenght: 1014bp
Host: Mammalian cells,lentivirus
Use(s): Protein Expression
Fragment type: ORF
Species: Virus
Prokaryotic resistance: Amp
Screening marker: Puro
Fluorescent labeling: Green
Promoter: EF1a
Replicon: pUC
Growth Strain(s): Stbl3
Growth Temperature: 37°C
Vector backbone: pLV3-EF1a-MCS-3×FLAG-PGK-CopGFP-Puro
Forward primer: M13F-72
Reverse primer:
Copy number: High
Induction:

pLV3-EF1a-CD47-scFv-3×FLAG-PGK-CopGFP-Puro plasmid Description

pLV3-EF1a-CD47-scFv-3×FLAG-PGK-CopGFP-Puro This plasmid is suitable for overexpression in mammalian cells. The plasmid contains the EF1a promoter, which can initiate the transcription of the CD47-scFv gene, promoting the C-terminal fusion expression of the CD47-scFv with a 3×FLAG tag, facilitating the detection, localization, and purification of the expressed fusion protein using FLAG antibodies. Additionally, the plasmid carries HIV-1 Ψ and 3'/5' LTR elements for lentivirus packaging, allowing for stable integration of the gene into host cells. The plasmid also carries the CopGFP green fluorescent protein, driven by the PGK promoter, for observing the expression and localization of the target gene. The plasmid contains a Puro resistance gene for selecting host cells expressing the target gene.

pLV3-EF1a-CD47-scFv-3×FLAG-PGK-CopGFP-Puro plasmid Sequence

1 ACGCGTGTAG TCTTATGCAA TACTCTTGTA GTCTTGCAAC ATGGTAACGA TGAGTTAGCA 61 ACATGCCTTA CAAGGAGAGA AAAAGCACCG TGCATGCCGA TTGGTGGAAG TAAGGTGGTA 121 CGATCGTGCC TTATTAGGAA GGCAACAGAC GGGTCTGACA TGGATTGGAC GAACCACTGA 181 ATTGCCGCAT TGCAGAGATA TTGTATTTAA GTGCCTAGCT CGATACAATA AACGGGTCTC 241 TCTGGTTAGA CCAGATCTGA GCCTGGGAGC TCTCTGGCTA ACTAGGGAAC CCACTGCTTA 301 AGCCTCAATA AAGCTTGCCT TGAGTGCTTC AAGTAGTGTG TGCCCGTCTG TTGTGTGACT 361 CTGGTAACTA GAGATCCCTC AGACCCTTTT AGTCAGTGTG GAAAATCTCT AGCAGTGGCG 421 CCCGAACAGG GACCTGAAAG CGAAAGGGAA ACCAGAGCTC TCTCGACGCA GGACTCGGCT 481 TGCTGAAGCG CGCACGGCAA GAGGCGAGGG GCGGCGACTG GTGAGTACGC CAAAAATTTT 541 GACTAGCGGA GGCTAGAAGG AGAGAGATGG GTGCGAGAGC GTCAGTATTA AGCGGGGGAG 601 AATTAGATCG CGATGGGAAA AAATTCGGTT AAGGCCAGGG GGAAAGAAAA AATATAAATT 661 AAAACATATA GTATGGGCAA GCAGGGAGCT AGAACGATTC GCAGTTAATC CTGGCCTGTT 721 AGAAACATCA GAAGGCTGTA GACAAATACT GGGACAGCTA CAACCATCCC TTCAGACAGG 781 ATCAGAAGAA CTTAGATCAT TATATAATAC AGTAGCAACC CTCTATTGTG TGCATCAAAG 841 GATAGAGATA AAAGACACCA AGGAAGCTTT AGACAAGATA GAGGAAGAGC AAAACAAAAG 901 TAAGACCACC GCACAGCAAG CGGCCACTGA TCTTCAGACC TGGAGGAGGA GATATGAGGG 961 ACAATTGGAG AAGTGAATTA TATAAATATA AAGTAGTAAA AATTGAACCA TTAGGAGTAG 1021 CACCCACCAA GGCAAAGAGA AGAGTGGTGC AGAGAGAAAA AAGAGCAGTG GGAATAGGAG 1081 CTTTGTTCCT TGGGTTCTTG GGAGCAGCAG GAAGCACTAT GGGCGCAGCC TCAATGACGC 1141 TGACGGTACA GGCCAGACAA TTATTGTCTG GTATAGTGCA GCAGCAGAAC AATTTGCTGA 1201 GGGCTATTGA GGCGCAACAG CATCTGTTGC AACTCACAGT CTGGGGCATC AAGCAGCTCC 1261 AGGCAAGAAT CCTGGCTGTG GAAAGATACC TAAAGGATCA ACAGCTCCTG GGGATTTGGG 1321 GTTGCTCTGG AAAACTCATT TGCACCACTG CTGTGCCTTG GAATGCTAGT TGGAGTAATA 1381 AATCTCTGGA ACAGATTGGA ATCACACGAC CTGGATGGAG TGGGACAGAG AAATTAACAA 1441 TTACACAAGC TTAATACACT CCTTAATTGA AGAATCGCAA AACCAGCAAG AAAAGAATGA 1501 ACAAGAATTA TTGGAATTAG ATAAATGGGC AAGTTTGTGG AATTGGTTTA ACATAACAAA 1561 TTGGCTGTGG TATATAAAAT TATTCATAAT GATAGTAGGA GGCTTGGTAG GTTTAAGAAT 1621 AGTTTTTGCT GTACTTTCTA TAGTGAATAG AGTTAGGCAG GGATATTCAC CATTATCGTT 1681 TCAGACCCAC CTCCCAACCC CGAGGGGACC CGACAGGCCC GAAGGAATAG AAGAAGAAGG 1741 TGGAGAGAGA GACAGAGACA GATCCATTCG ATTAGTGAAC GGATCTCGAC GGTTAACTTT 1801 TAAAAGAAAA GGGGGGATTG GGGGGTACAG TGCAGGGGAA AGAATAGTAG ACATAATAGC 1861 AACAGACATA CAAACTAAAG AATTACAAAA ACAAATTACA AAAATTCAAA ATTTTATCGA 1921 TACTAGTAAG GATCTGCGAT CGCTCCGGTG CCCGTCAGTG GGCAGAGCGC ACATCGCCCA 1981 CAGTCCCCGA GAAGTTGGGG GGAGGGGTCG GCAATTGAAC GGGTGCCTAG AGAAGGTGGC 2041 GCGGGGTAAA CTGGGAAAGT GATGTCGTGT ACTGGCTCCG CCTTTTTCCC GAGGGTGGGG 2101 GAGAACCGTA TATAAGTGCA GTAGTCGCCG TGAACGTTCT TTTTCGCAAC GGGTTTGCCG 2161 CCAGAACACA GCTGAAGCTT CGAGGGGCTC GCATCTCTCC TTCACGCGCC CGCCGCCCTA 2221 CCTGAGGCCG CCATCCACGC CGGTTGAGTC GCGTTCTGCC GCCTCCCGCC TGTGGTGCCT 2281 CCTGAACTGC GTCCGCCGTC TAGGTAAGTT TAAAGCTCAG GTCGAGACCG GGCCTTTGTC 2341 CGGCGCTCCC TTGGAGCCTA CCTAGACTCA GCCGGCTCTC CACGCTTTGC CTGACCCTGC 2401 TTGCTCAACT CTACGTCTTT GTTTCGTTTT CTGTTCTGCG CCGTTACAGA TCCAAGCTGT 2461 GACCGGCGCC TACTCTAGAG CTAGCGAATT CGCCACCATG GCTCTGCCTG TGACTGCCCT 2521 GCTGCTGCCA CTGGCCCTGC TGCTGCATGC CGCCAGGCCT GAGGTGCAGC TGCTGGAGAG 2581 CGGTGGCGGC CTTGTGCAGC CAGGCGGGTC TCTGAGGCTG TCCTGCGCCG CCAGCGGCTT 2641 TACATTCTCC TCTTACGCTA TGAGCTGGGT CCGGCAGGCC CCAGGCAAAG GCCTTGAGTG 2701 GGTGAGTGCC ATCAGTGGCA GCGGCGGCAG CACATACTAC GCTGACTCCG TGAAGGGAAG 2761 ATTTACCATT AGCAGAGATA ACAGCAAAAA CACCCTGTAC CTGCAGATGA ACAGCCTGCG 2821 AGCCGAAGAC ACCGCCGTGT ACTACTGCGC AAAATCCTAC GGCGCTTTCG ACTATTGGGG 2881 CCAGGGGACC CTGGTGACCG TGAGCAGCGG TGGAGGCGGA TCAGGAGGCG GTGGATCTGG 2941 CGGTGGAGGT TCAGATATCC AGATGACCCA GAGCCCCAGC TCTCTGAGTG CCTCCGTGGG 3001 GGACAGAGTG ACCATCACAT GTCAGGCCTC TCAGGACATC AATAAGTACC TGAACTGGTA 3061 TCAGCAGAAG CCCGGTAAGG CCCCAAAACT GCTGATTTAT GGCGCCAGCA GACTGGAGAC 3121 AGGCGTGCCC AGCAGATTCA GCGGATCAGG AAGCGGAACG GACTTTACAT TTACCATCTC 3181 CAGCCTGCAG CCCGAGGACA TCGCCACTTA CTACTGCCAG CAGAAACATC CAAGGGGCCC 3241 AAGAACCTTC GGCCAGGGAA CAAAAGTGGA AATCAAGGAA CAAAAACTCA TCTCAGAAGA 3301 GGATCTGATC AGCAACAGCG TGATGTACTT CAGCAGCGTG GTGCCCGTGC TGCAGAAAGT 3361 GAACAGCACC ACCACCAAGC CTGTGCTGAG AACCCCCAGC CCTGTGCACC CTACCGGCAG 3421 CAGCCTGAAG GGCACCGGCA TCTACATCTG GGCGCCCTTG GCCGGGACTT GTGGGGTCCT 3481 TCTCCTGTCA CTGGTTATCA CCCTTTACTG CGGATCCGAC TACAAAGACC ATGACGGTGA 3541 TTATAAAGAT CATGACATCG ACTACAAGGA TGACGATGAC AAGTAGCGGC CGCTGGGTAG 3601 GGGAGGCGCT TTTCCCAAGG CAGTCTGGAG CATGCGCTTT AGCAGCCCCG CTGGGCACTT 3661 GGCGCTACAC AAGTGGCCTC TGGCCTCGCA CACATTCCAC ATCCACCGGT AGGCGCCAAC 3721 CGGCTCCGTT CTTTGGTGGC CCCTTCGCGC CACCTTCTAC TCCTCCCCTA GTCAGGAAGT 3781 TCCCCCCCGC CCCGCAGCTC GCGTCGTGCA GGACGTGACA AATGGAAGTA GCACGTCTCA 3841 CTAGTCTCGT GCAGATGGAC AGCACCGCTG AGCAATGGAA GCGGGTAGGC CTTTGGGGCA 3901 GCGGCCAATA GCAGCTTTGC TCCTTCGCTT TCTGGGCTCA GAGGCTGGGA AGGGGTGGGT 3961 CCGGGGGCGG GCTCAGGGGC GGGCTCAGGG GCGGGGCGGG CGCCCGAAGG TCCTCCGGAG 4021 GCCCGGCATT CTGCACGCTT CAAAAGCGCA CGTCTGCCGC GCTGTTCTCC TCTTCCTCAT 4081 CTCCGGGCCT TTCGACCTGC AGCCCAAGCT TACCATGGAG AGCGACGAGA GCGGCCTGCC 4141 CGCCATGGAG ATCGAGTGCC GCATCACCGG CACCCTGAAC GGCGTGGAGT TCGAGCTGGT 4201 GGGCGGCGGA GAGGGCACCC CCAAGCAGGG CCGCATGACC AACAAGATGA AGAGCACCAA 4261 AGGCGCCCTG ACCTTCAGCC CCTACCTGCT GAGCCACGTG ATGGGCTACG GCTTCTACCA 4321 CTTCGGCACC TACCCCAGCG GCTACGAGAA CCCCTTCCTG CACGCCATCA ACAACGGCGG 4381 CTACACCAAC ACCCGCATCG AGAAGTACGA GGACGGCGGC GTGCTGCACG TGAGCTTCAG 4441 CTACCGCTAC GAGGCCGGCC GCGTGATCGG CGACTTCAAG GTGGTGGGCA CCGGCTTCCC 4501 CGAGGACAGC GTGATCTTCA CCGACAAGAT CATCCGCAGC AACGCCACCG TGGAGCACCT 4561 GCACCCCATG GGCGATAACG TGCTGGTGGG CAGCTTCGCC CGCACCTTCA GCCTGCGCGA 4621 CGGCGGCTAC TACAGCTTCG TGGTGGACAG CCACATGCAC TTCAAGAGCG CCATCCACCC 4681 CAGCATCCTG CAGAACGGGG GCCCCATGTT CGCCTTCCGC CGCGTGGAGG AGCTGCACAG 4741 CAACACCGAG CTGGGCATCG TGGAGTACCA GCACGCCTTC AAGACCCCCA TCGCCTTCGC 4801 CAGATCCCGC GCTCAGTCGT CCAATTCTGC CGTGGACGGC ACCGCCGGAC CCGGCTCCAC 4861 CGGATCTCGC GAGGGCAGAG GAAGTCTTCT AACATGCGGT GACGTGGAGG AGAATCCCGG 4921 CCCTATGACC GAGTACAAGC CCACGGTGCG CCTCGCCACC CGCGACGACG TCCCCAGGGC 4981 CGTACGCACC CTCGCCGCCG CGTTCGCCGA CTACCCCGCC ACGCGCCACA CCGTCGATCC 5041 GGACCGCCAC ATCGAGCGGG TCACCGAGCT GCAAGAACTC TTCCTCACGC GCGTCGGGCT 5101 CGACATCGGC AAGGTGTGGG TCGCGGACGA CGGCGCCGCG GTGGCGGTCT GGACCACGCC 5161 GGAGAGCGTC GAAGCGGGGG CGGTGTTCGC CGAGATCGGC CCGCGCATGG CCGAGTTGAG 5221 CGGTTCCCGG CTGGCCGCGC AGCAACAGAT GGAAGGCCTC CTGGCGCCGC ACCGGCCCAA 5281 GGAGCCCGCG TGGTTCCTGG CCACCGTCGG CGTCTCGCCC GACCACCAGG GCAAGGGTCT 5341 GGGCAGCGCC GTCGTGCTCC CCGGAGTGGA GGCGGCCGAG CGCGCCGGGG TGCCCGCCTT 5401 CCTGGAGACC TCCGCGCCCC GCAACCTCCC CTTCTACGAG CGGCTCGGCT TCACCGTCAC 5461 CGCCGACGTC GAGGTGCCCG AAGGACCGCG CACCTGGTGC ATGACCCGCA AGCCCGGTGC 5521 CTGAGTCGAC AATCAACCTC TGGATTACAA AATTTGTGAA AGATTGACTG GTATTCTTAA 5581 CTATGTTGCT CCTTTTACGC TATGTGGATA CGCTGCTTTA ATGCCTTTGT ATCATGCTAT 5641 TGCTTCCCGT ATGGCTTTCA TTTTCTCCTC CTTGTATAAA TCCTGGTTGC TGTCTCTTTA 5701 TGAGGAGTTG TGGCCCGTTG TCAGGCAACG TGGCGTGGTG TGCACTGTGT TTGCTGACGC 5761 AACCCCCACT GGTTGGGGCA TTGCCACCAC CTGTCAGCTC CTTTCCGGGA CTTTCGCTTT 5821 CCCCCTCCCT ATTGCCACGG CGGAACTCAT CGCCGCCTGC CTTGCCCGCT GCTGGACAGG 5881 GGCTCGGCTG TTGGGCACTG ACAATTCCGT GGTGTTGTCG GGGAAATCAT CGTCCTTTCC 5941 TTGGCTGCTC GCCTGTGTTG CCACCTGGAT TCTGCGCGGG ACGTCCTTCT GCTACGTCCC 6001 TTCGGCCCTC AATCCAGCGG ACCTTCCTTC CCGCGGCCTG CTGCCGGCTC TGCGGCCTCT 6061 TCCGCGTCTT CGCCTTCGCC CTCAGACGAG TCGGATCTCC CTTTGGGCCG CCTCCCCGCC 6121 TGGTACCTTT AAGACCAATG ACTTACAAGG CAGCTGTAGA TCTTAGCCAC TTTTTAAAAG 6181 AAAAGGGGGG ACTGGAAGGG CTAATTCACT CCCAACGAAA ATAAGATCTG CTTTTTGCTT 6241 GTACTGGGTC TCTCTGGTTA GACCAGATCT GAGCCTGGGA GCTCTCTGGC TAACTAGGGA 6301 ACCCACTGCT TAAGCCTCAA TAAAGCTTGC CTTGAGTGCT TCAAGTAGTG TGTGCCCGTC 6361 TGTTGTGTGA CTCTGGTAAC TAGAGATCCC TCAGACCCTT TTAGTCAGTG TGGAAAATCT 6421 CTAGCAGTAG TAGTTCATGT CATCTTATTA TTCAGTATTT ATAACTTGCA AAGAAATGAA 6481 TATCAGAGAG TGAGAGGAAC TTGTTTATTG CAGCTTATAA TGGTTACAAA TAAAGCAATA 6541 GCATCACAAA TTTCACAAAT AAAGCATTTT TTTCACTGCA TTCTAGTTGT GGTTTGTCCA 6601 AACTCATCAA TGTATCTTAT CATGTCTGGC TCTAGCTATC CCGCCCCTAA CTCCGCCCAG 6661 TTCCGCCCAT TCTCCGCCCC ATGGCTGACT AATTTTTTTT ATTTATGCAG AGGCCGAGGC 6721 CGCCTCGGCC TCTGAGCTAT TCCAGAAGTA GTGAGGAGGC TTTTTTGGAG GCCTAGACTT 6781 TTGCAGAGAC GGCCCAAATT CGTAATCATG GTCATAGCTG TTTCCTGTGT GAAATTGTTA 6841 TCCGCTCACA ATTCCACACA ACATACGAGC CGGAAGCATA AAGTGTAAAG CCTGGGGTGC 6901 CTAATGAGTG AGCTAACTCA CATTAATTGC GTTGCGCTCA CTGCCCGCTT TCCAGTCGGG 6961 AAACCTGTCG TGCCAGCTGC ATTAATGAAT CGGCCAACGC GCGGGGAGAG GCGGTTTGCG 7021 TATTGGGCGC TCTTCCGCTT CCTCGCTCAC TGACTCGCTG CGCTCGGTCG TTCGGCTGCG 7081 GCGAGCGGTA TCAGCTCACT CAAAGGCGGT AATACGGTTA TCCACAGAAT CAGGGGATAA 7141 CGCAGGAAAG AACATGTGAG CAAAAGGCCA GCAAAAGGCC AGGAACCGTA AAAAGGCCGC 7201 GTTGCTGGCG TTTTTCCATA GGCTCCGCCC CCCTGACGAG CATCACAAAA ATCGACGCTC 7261 AAGTCAGAGG TGGCGAAACC CGACAGGACT ATAAAGATAC CAGGCGTTTC CCCCTGGAAG 7321 CTCCCTCGTG CGCTCTCCTG TTCCGACCCT GCCGCTTACC GGATACCTGT CCGCCTTTCT 7381 CCCTTCGGGA AGCGTGGCGC TTTCTCATAG CTCACGCTGT AGGTATCTCA GTTCGGTGTA 7441 GGTCGTTCGC TCCAAGCTGG GCTGTGTGCA CGAACCCCCC GTTCAGCCCG ACCGCTGCGC 7501 CTTATCCGGT AACTATCGTC TTGAGTCCAA CCCGGTAAGA CACGACTTAT CGCCACTGGC 7561 AGCAGCCACT GGTAACAGGA TTAGCAGAGC GAGGTATGTA GGCGGTGCTA CAGAGTTCTT 7621 GAAGTGGTGG CCTAACTACG GCTACACTAG AAGGACAGTA TTTGGTATCT GCGCTCTGCT 7681 GAAGCCAGTT ACCTTCGGAA AAAGAGTTGG TAGCTCTTGA TCCGGCAAAC AAACCACCGC 7741 TGGTAGCGGT GGTTTTTTTG TTTGCAAGCA GCAGATTACG CGCAGAAAAA AAGGATCTCA 7801 AGAAGATCCT TTGATCTTTT CTACGGGGTC TGACGCTCAG TGGAACGAAA ACTCACGTTA 7861 AGGGATTTTG GTCATGAGAT TATCAAAAAG GATCTTCACC TAGATCCTTT TAAATTAAAA 7921 ATGAAGTTTT AAATCAATCT AAAGTATATA TGAGTAAACT TGGTCTGACA GTTACCAATG 7981 CTTAATCAGT GAGGCACCTA TCTCAGCGAT CTGTCTATTT CGTTCATCCA TAGTTGCCTG 8041 ACTCCCCGTC GTGTAGATAA CTACGATACG GGAGGGCTTA CCATCTGGCC CCAGTGCTGC 8101 AATGATACCG CGAGACCCAC GCTCACCGGC TCCAGATTTA TCAGCAATAA ACCAGCCAGC 8161 CGGAAGGGCC GAGCGCAGAA GTGGTCCTGC AACTTTATCC GCCTCCATCC AGTCTATTAA 8221 TTGTTGCCGG GAAGCTAGAG TAAGTAGTTC GCCAGTTAAT AGTTTGCGCA ACGTTGTTGC 8281 CATTGCTACA GGCATCGTGG TGTCACGCTC GTCGTTTGGT ATGGCTTCAT TCAGCTCCGG 8341 TTCCCAACGA TCAAGGCGAG TTACATGATC CCCCATGTTG TGCAAAAAAG CGGTTAGCTC 8401 CTTCGGTCCT CCGATCGTTG TCAGAAGTAA GTTGGCCGCA GTGTTATCAC TCATGGTTAT 8461 GGCAGCACTG CATAATTCTC TTACTGTCAT GCCATCCGTA AGATGCTTTT CTGTGACTGG 8521 TGAGTACTCA ACCAAGTCAT TCTGAGAATA GTGTATGCGG CGACCGAGTT GCTCTTGCCC 8581 GGCGTCAATA CGGGATAATA CCGCGCCACA TAGCAGAACT TTAAAAGTGC TCATCATTGG 8641 AAAACGTTCT TCGGGGCGAA AACTCTCAAG GATCTTACCG CTGTTGAGAT CCAGTTCGAT 8701 GTAACCCACT CGTGCACCCA ACTGATCTTC AGCATCTTTT ACTTTCACCA GCGTTTCTGG 8761 GTGAGCAAAA ACAGGAAGGC AAAATGCCGC AAAAAAGGGA ATAAGGGCGA CACGGAAATG 8821 TTGAATACTC ATACTCTTCC TTTTTCAATA TTATTGAAGC ATTTATCAGG GTTATTGTCT 8881 CATGAGCGGA TACATATTTG AATGTATTTA GAAAAATAAA CAAATAGGGG TTCCGCGCAC 8941 ATTTCCCCGA AAAGTGCCAC CTGACGTCTA AGAAACCATT ATTATCATGA CATTAACCTA 9001 TAAAAATAGG CGTATCACGA GGCCCTTTCG TCTCGCGCGT TTCGGTGATG ACGGTGAAAA 9061 CCTCTGACAC ATGCAGCTCC CGGAGACGGT CACAGCTTGT CTGTAAGCGG ATGCCGGGAG 9121 CAGACAAGCC CGTCAGGGCG CGTCAGCGGG TGTTGGCGGG TGTCGGGGCT GGCTTAACTA 9181 TGCGGCATCA GAGCAGATTG TACTGAGAGT GCACCATATG CGGTGTGAAA TACCGCACAG 9241 ATGCGTAAGG AGAAAATACC GCATCAGGCG CCATTCGCCA TTCAGGCTGC GCAACTGTTG 9301 GGAAGGGCGA TCGGTGCGGG CCTCTTCGCT ATTACGCCAG CTGGCGAAAG GGGGATGTGC 9361 TGCAAGGCGA TTAAGTTGGG TAACGCCAGG GTTTTCCCAG TCACGACGTT GTAAAACGAC 9421 GGCCAGTGCC AAGCTG

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Caution:
1.This product is FOR RESEARCH USE ONLY!
2.The item is lyophilized form, Please take the powder plasmid by centrifugation at 5000rpm/min for 1min. Add 20μl ddH2O in to the tube of plasmid.Take 2ul plasmid into 100 μ L corresponding competent cell, centrifugal then full coating on plate.
3.Shipping temperature is 2-8℃.