| 质粒名称: | pcDNA3.1(+)/Puro/FA/GFP-LL5BIP |
|---|---|
| 出品公司： | addgene |
| 目录编号: | 112829 |
| 存储实验室: | Tomasz Prószyński |
|
载体骨架基本信息 |
|
| 载体名称: | pcDNA3.1(+)/Puro |
| 载体出品公司: | Clontech |
| 空载体大小: | 5500bp |
| 完整质粒大小: | 6716bp |
| 载体修饰: | 人细胞色素c氧化酶亚基VIII（human cytochrome C oxidase subunit VIII） |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | Puromycin |
| 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: | LL5BIP |
| 物种来源: | M. musculus (mouse) |
| 标签/融合蛋白: | GFP (N terminal on insert) |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | 未知(未破坏) |
| 3’克隆位点: | 未知 (未破坏) |
|
文献引用方法 |
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| 材料和方法部分: | pcDNA3.1(+)/Puro/FA/GFP-LL5BIP was a gift from Tomasz Prószyński (Addgene plasmid # 112829 ; http://n2t.net/addgene:112829 ; RRID:Addgene_112829) |
| 参考文献部分: | LL5β interacts with uncharacterised protein 2410002F23Rik (LL5BIP). Niewiadomski P, Prószyński TJ. Matters v. 2, n. 11 (Nov 17, 2016) 10.19185/matters.201610000003 |
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
PuroFAGFP-LL5BIP质粒图谱.png)
载体简介
载体序列
LOCUS pcDNA3.1(+)/Puro/FA/GFP-LL5BIP 6716 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION transient/stable overexpression. ACCESSION . VERSION . KEYWORDS pcDNA3.1(+)/Puro/FA/GFP-LL5BIP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6716) TITLE 10.19185/matters.201610000003 REFERENCE 2 (bases 1 to 6716) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.biofeng.com FEATURES Location/Qualifiers source 1..6716 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 18..397 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 398..601 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 552..572 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 646..665 /label=T7 /note="T7 promoter, forward primer" promoter 646..664 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" CDS 718..1434 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" CDS 718..1434 /codon_start=1 /product="the original enhanced GFP (Yang et al., 1996)" /label=EGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(763..784) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(1024..1043) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 1371..1392 /label=EGFP-C /note="EGFP, forward primer" primer_bind complement(2345..2362) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 2351..2575 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 2621..3049 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2708..2727) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 2918..2939 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(3058..3078) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 3063..3392 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 3243..3378 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3305..3324 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3413..4012 /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(3413..3432) /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" primer_bind 3909..3929 /label=Puro-F /note="Puromycin resistance gene, forward primer" polyA_signal 4172..4293 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4209..4228) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4263..4282 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(4342..4358) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4342..4358) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4355..4377) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4366..4382 /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(4390..4420) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4435..4456 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(4573..4590) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4744..5332) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4824..4843) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5503..6363) /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 6126..6145 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6364..6468) /gene="bla" /label=AmpR promoter primer_bind complement(6543..6562) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" ORIGIN 1 ggccagatat acgcgttgac attgattatt gactagttat taatagtaat caattacggg 61 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc 121 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat 181 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc 241 ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga 301 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg 361 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat 421 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt 481 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc 541 cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc 601 tctctggcta actagagaac ccactgctta ctggcttatc gaaattaata cgactcacta 661 tagggagacc caagctggct agcgtttaaa cttaagcttg gtaccgagct cggatccatg 721 gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc 781 gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc 841 aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc 901 gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca catgaagcag 961 cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc 1021 aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg 1081 aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag 1141 ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc 1201 atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac 1261 cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac 1321 ctgagcaccc agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg 1381 ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagggccgg 1441 ccaaccatgg ctgagagagt gctggtgccc acccagatag gccgggggga ccgctactac 1501 acatacacgg agttgttggc tatctcacgg cgtttcaagc agaaccccaa tgagctcatg 1561 gtcacctgga tcctgcgggt gtatgaccag ggaggccctg ccctgtccct gaattctggg 1621 gagctggggc tgctgggtga cctcacccat gatgccatct ttaactaccg ctgcaaggcc 1681 ctgcgggggg ctggctgcca gactctcctg agctggctgc tgcaggcctg gcgtcagcgt 1741 tgggaatcct ccctgcattt tgaggccacc gagttaccct tcaggccctg gaccaccatg 1801 gaggaaggca tccagttggt gcgtgagctg ggcatgattg agtggatcta ccttgaccca 1861 gaagggcccg tggacctggc cccagaggat gtggccttca ctcaaggcct gcagcggcgc 1921 ctgctcacag cagccccctc tgagctgcgg ctttcactgg tcagcctgct ggtacgtggc 1981 atgacagtgt tggaggctgt gatggagatc cagactattg ctgacgtggg tctgctctgg 2041 cgccagagcc atccaggccg caccaagctc atgttggggc ccaacccaac tcgtaaggac 2101 ctcctaggct ggctgctcag ccacggtgta ccccgggagc aagtagacag acagcccacc 2161 aaggtactcc tagaactcta catcaaagaa gccaagcgca gtcgcggcca ccccaactat 2221 gggctgaacg aggagcagcc cccaccaccc ccttactccg accaggcctg tggggaagag 2281 caaccggtgc gtcatgacta ggcgcgcctc gagtctagag ggcccgttta aacccgctga 2341 tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct 2401 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca 2461 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag 2521 ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct 2581 gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca 2641 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta 2701 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt 2761 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac 2821 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt 2881 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga 2941 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg 3001 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga 3061 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa 3121 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca 3181 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc 3241 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt 3301 tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag 3361 gaggcttttt tggaggccta ggcttttgca aaaagctccc ggggccgcca ccatgaccga 3421 gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc ccccgggccg tacgcaccct 3481 cgccgccgcg ttcgccgact accccgccac gcgccacacc gtcgacccgg accgccacat 3541 cgagcgggtc accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa 3601 ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga 3661 agcgggggcg gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct 3721 ggccgcgcag caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg 3781 gttcctggcc accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt 3841 cgtgctcccc ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacctc 3901 cgcgccccgc aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga 3961 ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag cccggtgcct gattcgaaat 4021 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta 4081 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg 4141 ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta 4201 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag 4261 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag 4321 ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac 4381 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt 4441 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc 4501 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg 4561 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt 4621 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa 4681 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc 4741 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag 4801 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt 4861 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg 4921 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg 4981 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg 5041 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac 5101 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg 5161 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt 5221 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg 5281 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc 5341 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt 5401 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt 5461 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag 5521 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt 5581 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc 5641 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc 5701 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg 5761 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac 5821 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg 5881 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc 5941 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact 6001 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc 6061 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat 6121 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc 6181 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac 6241 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa 6301 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact 6361 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg 6421 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg 6481 aaaagtgcca cctgacgtcg acggatcggg agatctcccg atcccctatg gtgcactctc 6541 agtacaatct gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg 6601 gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac 6661 aattgcatga agaatctgct tagggttagg cgttttgcgc tgcttcgcga tgtacg //
