pCMV-GFP

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相关技术服务:质粒构建    基因合成    质粒大提

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pCMVGFP载体基本信息

质粒名称: pCMV-GFP、pCMVGFP
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
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
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
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)
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

pCMVGFP载体图谱质粒图谱和多克隆位点信息

pCMV-GFP载体图谱

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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