质粒名称: | pCMV-GFP、pCMVGFP |
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出品公司: | Addgene |
目录编号: | 11153 |
存储实验室: | Connie Cepko |
载体骨架基本信息 |
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载体名称: | pCAGEN |
载体出品公司: | Clontech |
空载体大小: | 3084 bp |
载体类型: | 哺乳细胞表达 |
载体抗性: | 氨苄霉素 |
生长温度: | 37 ℃ |
宿主菌: | DH5a |
拷贝数: | 高拷贝 |
插入基因信息 |
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插入基因名称: | CMV promoter-EGFP |
基因又名: | cytomegalovirus immediate early promoter |
插入大小: | 1400 bp |
启动子: | CMV |
克隆方法: | 限制性内切酶 |
5’克隆位点: | Spe I(已破坏) |
3’克隆位点: | NotI (未破坏) |
5’测序引物 | CMV-F |
3’测序引物 | SV40pA-R |
备注: | 本载体插入基因EGFP,来自pEGFP-N1载体。 |
文献引用方法 |
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材料和方法部分: |
pCMV-GFP was a gift from Connie Cepko (Addgene plasmid # 11153 ; http://n2t.net/addgene:11153 ; RRID:Addgene_11153)
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参考文献部分: | Electroporation and RNA interference in the rodent retina in vivo and in vitro. Matsuda T, Cepko CL. Proc Natl Acad Sci U S A. 2004 Jan 6. 101(1):16-22. 10.1073/pnas.2235688100 PubMed 14603031 |
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pCMVGFP载体基本信息
pCMVGFP载体图谱质粒图谱和多克隆位点信息
pCMVGFP载体简介
pCMVGFP载体序列
LOCUS pCMV-GFP 4479 bp ds-DNA circular SYN 29-AUG-2023 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4479) AUTHORS biofeng.com TITLE Direct Submission JOURNAL Exported Tuesday, Aug 29, 2023 from SnapGene 4.3.6 to Vector NTI(R) format https://www.biofeng.com COMMENT VNTDATE|1031011200| COMMENT VNTDBDATE|1031011200| COMMENT VNTNAME|pCMV-GFP.dna| COMMENT VNTAUTHORNAME|biofeng.com| FEATURES Location/Qualifiers enhancer 58..361 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 362..565 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" misc_feature 588..668 /label=MCS /note="multiple cloning site" regulatory 670..679 /regulatory_class="other" /label=Kozak sequence /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 676..1395 /codon_start=1 /product="enhanced GFP" /label=EGFP /note="mammalian codon-optimized" polyA_signal 1547..1602 /label=beta-globin poly(A) signal /note="rabbit beta-globin polyadenylation signal (Gil and Proudfoot, 1987)" primer_bind complement(1963..1979) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" protein_bind 1987..2003 /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(2011..2041) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 2056..2077 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 2135..2331 /label=SV40 promoter /note="SV40 early promoter" rep_origin 2182..2317 /label=SV40 ori /note="SV40 origin of replication" polyA_signal 2337..2471 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" rep_origin complement(2710..3298) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(3469..4329) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" promoter complement(4330..4434) /gene="bla" /label=AmpR promoter ORIGIN 1 ctagtaatag taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt 61 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac 121 gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg 181 ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag 241 tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat 301 gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat 361 ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt 421 tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga 481 ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg 541 gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct agcgctaccg 601 gactcagatc tcgagctcaa gcttcgaatt ctgcagtcga cggtaccgcg ggcccgggat 661 ccaccggtcg ccaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc 721 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag 781 ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc 841 gtgccctggc ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac 901 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag 961 gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc 1021 gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc 1081 aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc 1141 gacaagcaga agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc 1201 agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg 1261 ctgcccgaca accactacct gagcacccag tccgccctga gcaaagaccc caacgagaag 1321 cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac 1381 gagctgtaca agtaaagcgg ccgcactcct caggtgcagg ctgcctatca gaaggtggtg 1441 gctggtgtgg ccaatgccct ggctcacaaa taccactgag atctttttcc ctctgccaaa 1501 aattatgggg acatcatgaa gccccttgag catctgactt ctggctaata aaggaaattt 1561 attttcattg caatagtgtg ttggaatttt ttgtgtctct cactcggaag gacatatggg 1621 agggcaaatc atttaaaaca tcagaatgag tatttggttt agagtttggc aacatatgcc 1681 catatgctgg ctgccatgaa caaaggttgg ctataaagag gtcatcagta tatgaaacag 1741 ccccctgctg tccattcctt attccataga aaagccttga cttgaggtta gatttttttt 1801 atattttgtt ttgtgttatt tttttcttta acatccctaa aattttcctt acatgtttta 1861 ctagccagat ttttcctcct ctcctgacta ctcccagtca tagctgtccc tcttctctta 1921 tggagatccc tcgacctgca gcccaagctt ggcgtaatca tggtcatagc tgtttcctgt 1981 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa 2041 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc 2101 tttccagtcg ggaaacctgt cgtgccagcg gatccgcatc tcaattagtc agcaaccata 2161 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg 2221 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag 2281 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctaact 2341 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata 2401 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc 2461 atgtctggat ccgctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat 2521 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg 2581 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc 2641 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt 2701 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag 2761 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc 2821 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc 2881 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt 2941 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt 3001 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc 3061 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa 3121 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa 3181 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg 3241 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga 3301 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg 3361 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg 3421 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt 3481 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact 3541 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat 3601 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg 3661 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg 3721 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat 3781 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc 3841 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt 3901 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc 3961 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga 4021 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc 4081 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa 4141 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta 4201 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg 4261 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg 4321 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat 4381 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt 4441 tccccgaaaa gtgccacctg ggtcgacatt gattattga //
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