质粒名称: | pBIR-GFP |
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出品公司: | 爱迪基因 |
目录编号: | 49807 |
存储实验室: | Thanos Halazonetis |
载体骨架基本信息 |
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载体名称: | pgkPURO |
载体抗性: | 氨苄青霉素 |
载体修饰: | GFP-IsceI:2037-2576;IsceI:2577-2594;嘌呤霉素:3858-3260;iGFP:5750-5169;poly-A信号:5015-4981 |
载体类型: | 哺乳细胞表达 |
筛选标记: | 嘌呤霉素 |
生长条件: | 37 ℃ |
插入基因信息 |
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插入基因名称: | GFP |
启动子 | beta-actin |
克隆方法: | 限制性内切酶 |
5' 克隆位点: | bbsI(未破坏) |
3' 克隆位点: | aflII (未破坏) |
5' 测序引物 | gatcaggcagagcaggaacctg |
3' 测序引物 | cctgaagaacgagatcagcagcctctgt |
文献引用方法 | |
材料和方法部分: | pBIR-GFP was a gift from Thanos Halazonetis (Addgene plasmid # 49807 ; http://n2t.net/addgene:49807 ; RRID:Addgene_49807) |
买家导航
pBIRGFP载体基本信息
pBIRGFP载体图谱质粒图谱和多克隆位点信息
pBIRGFP载体简介
pBIRGFP载体序列
LOCUS pBIR-GFP 8266 bp ds-DNA circular SYN 06-AUG-2018 DEFINITION In vivo GFP based system to measure break induced replication (BIR) repair efficiency. expression of I-SceI generates a double strand break that when repaired by BIR mechanism will restore GFP. . ACCESSION . VERSION . KEYWORDS pBIR-GFP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8266) TITLE pBIR-GFP JOURNAL Unpublished REFERENCE 2 (bases 1 to 8266) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8266 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 4..383 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 386..661 /label=chicken beta-actin promoter intron 662..1670 /label=chimeric intron /note="chimera between introns from chicken beta-actin and rabbit beta-globin" primer_bind 1594..1616 /label=pCAGGS-5 /note="Chimeric intron in CAG promoter, forward primer" primer_bind 1678..1697 /label=pCAG-F /note="Rabbit beta-globin intron, for pCAG plasmids, forward primer" CDS 1755..1775 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 1797..1817 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" regulatory 2031..2040 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 2037..2555 /codon_start=1 /product="N-terminal fragment of mVenus for use in bimolecular fluorescence complementation (BiFC) (Kodama and Hu, 2010)" /label=VN173 /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIE" primer_bind complement(2082..2103) /label=EGFP-N /note="EGFP, reverse primer" CDS complement(3259..3858) /codon_start=1 /gene="pac from Streptomyces alboniger" /product="puromycin N-acetyltransferase" /label=PuroR /note="confers resistance to puromycin" /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA" primer_bind complement(3342..3362) /label=Puro-F /note="Puromycin resistance gene, forward primer" primer_bind 3839..3858 /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" promoter complement(3877..4374) /label=PGK promoter /note="mouse phosphoglycerate kinase 1 promoter" primer_bind complement(3925..3944) /label=mPGK-F /note="Mouse PGK promoter, forward primer" primer_bind 4316..4339 /label=MSCV-rev /note="Murine stem cell virus, reverse primer" primer_bind complement(4462..4481) /label=Bglob-pA-R /note="Rabbit beta-globin polyA region, reverse primer" polyA_site 4527..4582 /label=-beta-globin poly(A) signal /note="rabbit beta-globin polyadenylation signal" primer_bind complement(4581..4600) /label=rbglobpA-R /note="Rabbit beta-globin polyA, reverse primer. Also called rb-glob-pA-term-R" polyA_signal 4965..5046 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5214..5235) /label=EGFP-C /note="EGFP, forward primer" primer_bind complement(5769..5785) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5769..5785) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(5782..5804) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 5793..5809 /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(5817..5847) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5862..5883 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 5942..6137 /label=SV40 promoter /note="SV40 early promoter" rep_origin 5988..6123 /label=SV40 ori /note="SV40 origin of replication" primer_bind 6050..6069 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" polyA_signal 6143..6277 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(6345..6362) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(6516..7104) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(6596..6615) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(7275..8135) /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 7898..7917 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(8136..8240) /gene="bla" /label=AmpR promoter ORIGIN 1 gtcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata 61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc 121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag 181 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac 241 atcaagtgta tcatatgcca agtccgcccc ctattgacgt caatgacggt aaatggcccg 301 cctggcatta tgcccagtac atgaccttac gggactttcc tacttggcag tacatctacg 361 tattagtcat cgctattacc atgggtcgag gtgagcccca cgttctgctt cactctcccc 421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca 481 gcgatggggg cggggggggg gggggcgcgc gccaggcggg gcggggcggg gcgaggggcg 541 gggcggggcg aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt 601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc 661 gggagtcgct gcgttgcctt cgccccgtgc cccgctccgc gccgcctcgc gccgcccgcc 721 ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc 781 gggctgtaat tagcgcttgg tttaatgacg gctcgtttct tttctgtggc tgcgtgaaag 841 ccttaaaggg ctccgggagg gccctttgtg cgggggggag cggctcgggg ggtgcgtgcg 901 tgtgtgtgtg cgtggggagc gccgcgtgcg gcccgcgctg cccggcggct gtgagcgctg 961 cgggcgcggc gcggggcttt gtgcgctccg cgtgtgcgcg aggggagcgc ggccgggggc 1021 ggtgccccgc ggtgcggggg ggctgcgagg ggaacaaagg ctgcgtgcgg ggtgtgtgcg 1081 tgggggggtg agcagggggt gtgggcgcgg cggtcgggct gtaacccccc cctgcacccc 1141 cctccccgag ttgctgagca cggcccggct tcgggtgcgg ggctccgtgc ggggcgtggc 1201 gcggggctcg ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg cggggcgggg 1261 ccgcctcggg ccggggaggg ctcgggggag gggcgcggcg gccccggagc gccggcggct 1321 gtcgaggcgc ggcgagccgc agccattgcc ttttatggta atcgtgcgag agggcgcagg 1381 gacttccttt gtcccaaatc tggcggagcc gaaatctggg aggcgccgcc gcaccccctc 1441 tagcgggcgc gggcgaagcg gtgcggcgcc ggcaggaagg aaatgggcgg ggagggcctt 1501 cgtgcgtcgc cgcgccgccg tccccttctc catctccagc ctcggggctg ccgcaggggg 1561 acggctgcct tcggggggga cggggcaggg cggggttcgg cttctggcgt gtgaccggcg 1621 gctctagagc ctctgctaac catgttcatg ccttcttctt tttcctacag ctcctgggca 1681 acgtgctggt tattgtgctg tctcatcatt ttggcaaaga attcagatcc gccgccacta 1741 tgggatcaag atcgccaaaa aagaagagaa aggtgccgaa gaagcatgca gcaccaccaa 1801 aaaaaaaacg aaaagtagaa gacccacgag gcaacaccag cggcgtgctg agcaccccca 1861 aggccaagag ggccaagcac ccccccggca ccgagaagcc caggagcagg agccagagcg 1921 agcagcccgc cacctgcccc atctgctacg ccgtgatcag gcagagcagg aacctgagga 1981 ggcacctgga gctgaggcac ttcgccaagc ccggcgtgga tccaccggtc gccaccatgg 2041 tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg 2101 acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca 2161 agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg 2221 tgaccaccct gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc 2281 acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca 2341 aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga 2401 accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc 2461 tggagtacaa ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca 2521 tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgcctagg 2581 gataacaggg taatcttaag gcgtaaattg taagcgttaa tctgaacctt ctgatggaat 2641 tagaacttgg caaaacaata ctgagaatga agtgtatgtg gaacagaggc tgctgatctc 2701 gttcttcagg ctatgaaact gacacatttg gaaaccacag tacttagaac cacaaagtgg 2761 gaatcaagag aaaaacaatg atcccacgag agatctatag atctatagat catgagtggg 2821 aggaatgagc tggcccttaa tttggttttg cttgtttaaa ttatgatatc caactatgaa 2881 acattatcat aaagcaatag taaagagcct tcagtaaaga gcaggcattt atctaatccc 2941 accccacccc cacccccgta gctccaatcc ttccattcaa aatgtaggta ctctgttctc 3001 acccttctta acaaagtatg acaggaaaaa cttccatttt agtggacatc tttattgttt 3061 aatagatcat caatttcgat ccgctcctgg gcaccgaact gcgccgcgtg ttcagcaggg 3121 tcggcgtgtt cggtgtgtcc cccgcggtgg gcctcggggg cgggtgcggg gtcggcgggg 3181 ccgccccggg tggcttcggt cggagccatg gggtcgtgcg ctcctttcgg tcgggcgctg 3241 cgggtcgtgg ggcgggcgtc aggcaccggg cttgcgggtc atgcaccagg tgcgcggtcc 3301 ttcgggcacc tcgacgtcgg cggtgacggt gaagccgagc cgctcgtaga aggggaggtt 3361 gcggggcgcg gaggtctcca ggaaggcggg caccccggcg cgctcggccg cctccactcc 3421 ggggagcacg acggcgctgc ccagaccctt gccctggtgg tcgggcgaga cgccgacggt 3481 ggccaggaac cacgcgggct ccttgggccg gtgcggcgcc aggaggcctt ccatctgttg 3541 ctgcgcggcc agccgggaac cgctcaactc ggccatgcgc gggccgatct cggcgaacac 3601 cgcccccgct tcgacgctct ccggcgtggt ccagaccgcc accgcggcgc cgtcgtccgc 3661 gacccacacc ttgccgatgt cgagcccgac gcgcgtgagg aagagttctt gcagctcggt 3721 gacccgctcg atgtggcggt ccgggtcgac ggtgtggcgc gtggcggggt agtcggcgaa 3781 cgcggcggcg agggtgcgta cggcccgggg gacgtcgtcg cgggtggcga ggcgcaccgt 3841 gggcttgtac tcggtcatgg tggcggctgg atcggtcgaa aggcccggag atgaggaaga 3901 ggagaacagc gcggcagacg tgcgcttttg aagcgtgcag aatgccgggc ctccggagga 3961 ccttcgggcg cccgccccgc ccctgagccc gcccctgagc ccgcccccgg acccacccct 4021 tcccagcctc tgagcccaga aagcgaagga gcaaagctgc tattggccgc tgccccaaag 4081 gcctacccgc ttccattgct cagcggtgct gtccatctgc acgagactag tgagacgtgc 4141 tacttccatt tgtcacgtcc tgcacgacgc gagctgcggg gcggggggga acttcctgac 4201 taggggagga gtagaaggtg gcgcgaaggg gccaccaaag aacggagccg gttggcgcct 4261 accggtggat gtggaatgtg tgaggccaga ggccacttgt gtagcgccaa gtgcccagcg 4321 gggctgctaa agcgcatgct ccagactgcc ttgggaaaag cgcctcccct acccggtaga 4381 attcactcct caggtgcagg ctgcctatca gaaggtggtg gctggtgtgg ccaatgccct 4441 ggctcacaaa taccactgag atctttttcc ctctgccaaa aattatgggg acatcatgaa 4501 gccccttgag catctgactt ctggctaata aaggaaattt attttcattg caatagtgtg 4561 ttggaatttt ttgtgtctct cactcggaag gacatatggg agggcaaatc atttaaaaca 4621 tcagaatgag tatttggttt agagtttggc aacatatgcc atatgctggc tgccatgaac 4681 aaaggtggct ataaagaggt catcagtata tgaaacagcc ccctgctgtc cattccttat 4741 tccatagaaa agccttgact tgaggttaga ttttttttat attttgtttt gtgttatttt 4801 tttctttaac atccctaaaa ttttccttac atgttttact agccagattt ttcctcctct 4861 cctgactact cccagtcata gctgtccctc ttctcttatg aagatccctc gacctgcagc 4921 ccaagcttat acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc 4981 tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa 5041 caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga ggtgtgggag 5101 gttttttaaa gcaagtaaaa cctctacaaa tgtggtatgg ctgattatga tctagagtcg 5161 cggccgcttt acttgtacag ctcgtccatg ccgagagtga tcccggcggc ggtcacgaac 5221 tccagcagga ccatgtgatc gcgcttctcg ttggggtctt tgctcagggc ggactgggtg 5281 ctcaggtagt ggttgtcggg cagcagcacg gggccgtcgc cgatgggggt gttctgctgg 5341 tagtggtcgg cgagctgcac gctgccgtcc tcgatgttgt ggcggatctt gaagttcacc 5401 ttgatgccgt tcttctgctt gtcggccatg atatagacgt tgtggctgtt gtagttgtac 5461 tccagcttgt gccccaggat gttgccgtcc tccttgaagt cgatgccctt cagctcgatg 5521 cggttcacca gggtgtcgcc ctcgaacttc acctcggcgc gggtcttgta gttgccgtcg 5581 tccttgaaga agatggtgcg ctcctggacg tagccttcgg gcatggcgga cttgaagaag 5641 tcgtgctgct tcatgtggtc ggggtagcgg ctgaagcact gcacgccgta ggtcagggtg 5701 gtcacgaggg tgggccaggg cacgggcagc ttgccggtgg tgcagatgaa aagcttggcg 5761 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac 5821 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca 5881 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagcggatc 5941 cgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa 6001 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag 6061 aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag 6121 gcctaggctt ttgcaaaaag ctaacttgtt tattgcagct tataatggtt acaaataaag 6181 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt 6241 gtccaaactc atcaatgtat cttatcatgt ctggatccgc tgcattaatg aatcggccaa 6301 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg 6361 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg 6421 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag 6481 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac 6541 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga 6601 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt 6661 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca aagctcacgc 6721 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc 6781 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta 6841 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat 6901 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca 6961 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct 7021 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt 7081 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct 7141 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc 7201 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa 7261 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta 7321 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc 7381 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat 7441 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta 7501 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt 7561 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt 7621 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg 7681 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc 7741 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc 7801 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg 7861 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga 7921 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta 7981 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct 8041 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag 8101 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga 8161 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat 8221 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctg //