质粒名称: | pLL3.7m-Clover-Geminin(1-110)-IRES-mKO2-Cdt(30-120) |
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出品公司: | 爱迪基因 |
目录编号: | 83841 |
存储实验室: | Michael Lin |
载体骨架基本信息 |
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载体名称: | pLL3.7m |
载体抗性: | 氨苄青霉素 |
空载体大小: | 6500 bp |
完整质粒大小: | 9036 bp |
载体类型: | 哺乳动物表达,慢病毒 |
高拷贝/低拷贝: | 高拷贝 |
生长条件: | 37 ℃ |
插入基因信息 |
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插入基因名称: | Clover-Geminin(1-110)-IRES-mKO2-Cdt(30-120) |
物种来源: | H. sapiens (human), Synthetic |
插入大小: | 2649 bp |
启动子 | CMV |
克隆方法: | 限制性内切酶 |
5' 克隆位点: | NheI(未破坏) |
3' 克隆位点: | EcoRI (未破坏) |
5' 测序引物 | agctggtttagtgaaccgtcagatc |
3' 测序引物 | ggaaccggaacccttaaaca |
文献引用方法 |
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材料和方法部分: | pLL3.7m-Clover-Geminin(1-110)-IRES-mKO2-Cdt(30-120) was a gift from Michael Lin (Addgene plasmid # 83841 ; http://n2t.net/addgene:83841 ; RRID:Addgene_83841) |
参考文献部分: | Fluorescent indicators for simultaneous reporting of all four cell cycle phases. Bajar BT, Lam AJ, Badiee RK, Oh YH, Chu J, Zhou XX, Kim N, Kim BB, Chung M, Yablonovitch AL, Cruz BF, Kulalert K, Tao JJ, Meyer T, Su XD, Lin MZ. Nat Methods. 2016 Oct 31. doi: 10.1038/nmeth.4045. 10.1038/nmeth.4045 PubMed 27798610 |
pLL3.7m-Clover-Geminin(1-110)-IRES-mKO2-Cdt(30-120)
买家导航
载体基本信息
载体图谱质粒图谱和多克隆位点信息
载体简介
载体序列
LOCUS pLL3.7m-Clover-Geminin(1-110)-IRES-mKO2-Cdt(30-120) 9033 bp ds-DNA circular SYN 06-AUG-2018 DEFINITION Fluorescent probe for G1/S transition and M/G1 transition. ACCESSION . VERSION . KEYWORDS pLL3.7m-Clover-Geminin(1-110)-IRES-mKO2-Cdt(30-120) SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9033) AUTHORS Bajar BT, Lam AJ, Badiee RK, Oh YH, Chu J, Zhou XX, Kim N, Kim BB, Chung M, Yablonovitch AL, Cruz BF, Kulalert K, Tao JJ, Meyer T, Su XD, Lin MZ TITLE Fluorescent indicators for simultaneous reporting of all four cell cycle phases. JOURNAL Nat Methods. 2016 Oct 31. doi: 10.1038/nmeth.4045. PUBMED 27798610 REFERENCE 2 (bases 1 to 9033) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9033 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 63..366 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 368..566 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" LTR 584..764 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 811..936 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 1433..1666 /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 2193..2310 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" primer_bind 2600..2620 /label=mU6-F /note="Mouse U6 promoter, forward primer" primer_bind 2698..2714 /label=KS primer /note="common sequencing primer, one of multiple similar variants" protein_bind 2756..2789 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." enhancer 2905..3208 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 3209..3412 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" CDS 3444..4160 /codon_start=1 /product="bright green-yellow fluorescent protein derived from GFP (Lam et al., 2012)" /label=Clover /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL KFICTTGKLPVPWPTLVTTFGYGVACFSRYPDHMKQHDFFKSAMPEGYVQERTISFKDD GTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIK ANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSHQSALSKDPNEKRDHMVLL EFVTAAGITHGMDELYK" primer_bind complement(3489..3510) /label=EGFP-N /note="EGFP, reverse primer" primer_bind 4097..4118 /label=EGFP-C /note="EGFP, forward primer" misc_feature 4525..5097 /label=IRES /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" primer_bind complement(4692..4709) /label=IRES reverse /note="IRES internal ribosome entry site, reverse primer. Also called pCDH-rev" primer_bind 4919..4938 /label=IRES-F /note="IRES internal ribosome entry site, forward primer" protein_bind 6120..6153 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." misc_feature 6209..6797 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(6262..6282) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(6680..6691) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" primer_bind complement(6800..6816) /label=KS primer /note="common sequencing primer, one of multiple similar variants" LTR 7039..7219 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" rep_origin complement(7281..7869) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(7361..7380) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(8040..8900) /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" primer_bind 8663..8682 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(8901..9005) /gene="bla" /label=AmpR promoter ORIGIN 1 actagttatt aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc 61 cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca 121 ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt 181 caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg 241 ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag 301 tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt 361 accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg 421 ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa 481 cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt 541 gtacggtggg aggtctatat aagcagcgcg ttttgcctgt actgggtctc tctggttaga 601 ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata 661 aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta 721 gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg 781 gacttgaaag cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa 841 gcgcgcacgg caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc 901 ggaggctaga aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga 961 tcgcgatggg aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat 1021 atagtatggg caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca 1081 tcagaaggct gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa 1141 gaacttagat cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag 1201 ataaaagaca ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc 1261 accgcacagc aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca 1321 attggagaag tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac 1381 ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt 1441 tgttccttgg gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga 1501 cggtacaggc cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg 1561 ctattgaggc gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg 1621 caagaatcct ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt 1681 gctctggaaa actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat 1741 ctctggaaca gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt 1801 acacaagctt aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac 1861 aagaattatt ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt 1921 ggctgtggta tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag 1981 tttttgctgt actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc 2041 agacccacct cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg 2101 gagagagaga cagagacaga tccattcgat tagtgaacgg atcggcactg cgtgcgccaa 2161 ttctgcagac aaatggcagt attcatccac aattttaaaa gaaaaggggg gattgggggg 2221 tacagtgcag gggaaagaat agtagacata atagcaacag acatacaaac taaagaatta 2281 caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt acagggacag cagagatcca 2341 gtttggttag taccgggccc gctctagaga tccgacgcgc catctctagg cccgcgccgg 2401 ccccctcgca cagacttgtg ggagaagctc ggctactccc ctgccccggt taatttgcat 2461 ataatatttc ctagtaacta tagaggctta atgtgcgata aaagacagat aatctgttct 2521 ttttaatact agctacattt tacatgatag gcttggattt ctataagaga tacaaatact 2581 aaattattat tttaaaaaac agcacaaaag gaaactcacc ctaactgtaa agtaattgtg 2641 tgttttgaga ctataaatat cccttggaga aaagccttgt taacgcgcgg tgaccctcga 2701 ggtcgacggt atcgataagc tcgcttcacg agattccagc aggtcgaggg acctaataac 2761 ttcgtatagc atacattata cgaagttata ttaagggttc caagcttaag cggccgcgtg 2821 gataaccgta ttaccgccat gcattagtta ttaatagtaa tcaattacgg ggtcattagt 2881 tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg 2941 accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc 3001 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc 3061 agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg 3121 gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat 3181 ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg 3241 tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag 3301 tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt 3361 gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctggtttagt 3421 gaaccgtcag atccgctagc accatggtga gcaagggcga ggagctgttc accggggtgg 3481 tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc gtccgcggcg 3541 agggcgaggg cgatgccacc aacggcaagc tgaccctgaa gttcatctgc accaccggca 3601 agctgcccgt gccctggccc accctcgtga ccaccttcgg ctacggcgtg gcctgcttca 3661 gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct 3721 acgtccagga gcgcaccatc tctttcaagg acgacggtac ctacaagacc cgcgccgagg 3781 tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg 3841 aggacggcaa catcctgggg cacaagctgg agtacaactt caacagccac aacgtctata 3901 tcacggccga caagcagaag aacggcatca aggctaactt caagatccgc cacaacgttg 3961 aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc 4021 ccgtgctgct gcccgacaac cactacctga gccatcagtc cgccctgagc aaagacccca 4081 acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg attacacatg 4141 gcatggacga gctgtacaag gatatccatg cacactggcg gccgctcgag atgaatccca 4201 gtatgaagca gaaacaagaa gaaatcaaag agaatataaa gaatagttct gtcccaagaa 4261 gaactctgaa gatgattcag ccttctgcat ctggatctct tgttggaaga gaaaatgagc 4321 tgtccgcagg cttgtccaaa aggaaacatc ggaatgacca cttaacatct acaacttcca 4381 gccctggggt tattgtccca gaatctagtg aaaataaaaa tcttggagga gtcacccagg 4441 agtcatttga tcttatgatt aaagaaaatc catcctctca gtattggaag gaagtggcag 4501 aaaaacggag aaaggcgctg taagcccctc tccctccccc ccccctaacg ttactggccg 4561 aagccgcttg gaataaggcc ggtgtgcgtt tgtctatatg ttattttcca ccatattgcc 4621 gtcttttggc aatgtgaggg cccggaaacc tggccctgtc ttcttgacga gcattcctag 4681 gggtctttcc cctctcgcca aaggaatgca aggtctgttg aatgtcgtga aggaagcagt 4741 tcctctggaa gcttcttgaa gacaaacaac gtctgtagcg accctttgca ggcagcggaa 4801 ccccccacct ggcgacaggt gcctctgcgg ccaaaagcca cgtgtataag atacacctgc 4861 aaaggcggca caaccccagt gccacgttgt gagttggata gttgtggaaa gagtcaaatg 4921 gctctcctca agcgtattca acaaggggct gaaggatgcc cagaaggtac cccattgtat 4981 gggatctgat ctggggcctc ggtgcacatg ctttacatgt gtttagtcga ggttaaaaaa 5041 cgtctaggcc ccccgaacca cggggacgtg gttttccttt gaaaaacacg atgataatgg 5101 ccacagaatt catggtgagc gtgatcaagc ccgagatgaa gatgaggtac tacatggacg 5161 gctccgtcaa tgggcatgag ttcacaatcg agggtgaggg cacaggcaga ccttacgagg 5221 gacatcagga gatgacactg cgcgtcacaa tggccgaggg cgggccaatg cctttcgcct 5281 tcgacctggt gtcccacgtg ttctgttacg gccacagagt ttttaccaag tacccagaag 5341 agatcccaga ctatttcaag caggcctttc ctgagggcct gtcctgggag aggtccctgg 5401 agttcgagga cggcggctcc gcctccgtga gcgcccacat cagcctgagg ggcaacacct 5461 tctaccacaa gtccaagttc accggcgtga acttccccgc cgacggcccc atcatgcaga 5521 accagagcgt ggactgggag ccctccaccg agaagatcac cgccagcgac ggcgtgctga 5581 agggcgacgt gaccatgtac ctgaagctgg agggcggcgg caaccacaag tgccagatga 5641 agaccaccta caaggccgcc aaggagatcc tggagatgcc cggcgaccac tacatcggcc 5701 acaggctggt gaggaagacc gagggcaaca tcaccgagca ggtggaggac gccgtggccc 5761 actactcaat gttgccttcc ctgggcgata tccatgcaca ctggcggccg ctcgagccca 5821 gccccgccag gcccgcactc cgcgccccgg cctccgctac cagtggcagc cgcaagcgcg 5881 cccgcccgcc cgccgccccc ggacgcgacc aggccaggcc accggcccgc aggagactgc 5941 ggctgtcggt ggacgaggtt tccagcccca gtacccccga ggccccagac atcccagcct 6001 gcccttctcc gggccagaag ataaagaaat ccaccccggc agcaggtcag ccgccccacc 6061 tgacatccgc gcaggaccag gacaccatct gagtttaaac gaattcgtcg agggacctaa 6121 taacttcgta tagcatacat tatacgaagt tatacatgtt taagggttcc ggttccacta 6181 ggtacaattc gatatcaagc ttatcgataa tcaacctctg gattacaaaa tttgtgaaag 6241 attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat 6301 gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc 6361 ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg 6421 cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct 6481 ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct 6541 tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg 6601 gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac 6661 gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct 6721 gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct 6781 ttgggccgcc tccccgcatc gataccgtcg acctcgatcg agacctagaa aaacatggag 6841 caatcacaag tagcaataca gcagctacca atgctgattg tgcctggcta gaagcacaag 6901 aggaggagga ggtgggtttt ccagtcacac ctcaggtacc tttaagacca atgacttaca 6961 aggcagctga cgcgtgattt taaaagaaaa ggggggactg gaagggctaa ttcactccca 7021 ctgctttttg cctgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct 7081 ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta 7141 gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca 7201 gtgtggaaaa tctctagcag catgtgagca aaaggccagc aaaaggccag gaaccgtaaa 7261 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat 7321 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc 7381 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc 7441 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt 7501 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac 7561 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg 7621 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca 7681 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc 7741 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa 7801 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa 7861 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac 7921 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta 7981 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt 8041 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata 8101 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc 8161 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac 8221 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag 8281 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac 8341 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc 8401 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg 8461 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc 8521 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct 8581 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc 8641 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc 8701 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc 8761 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc 8821 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca 8881 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt 8941 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt 9001 ccgcgcacat ttccccgaaa agtgccacct gac //
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