质粒名称: | pLS-SceI, pLS-Scel |
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出品公司: | Addgene |
目录编号: | 137725 |
存储实验室: | Nadav Ahituv |
载体骨架基本信息 |
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载体名称: | pLS-SceI |
空载体大小: | 7811 bp |
载体类型: | 慢病毒载体 |
载体抗性: | 氨苄霉素100 μg/mL |
生长温度: | 37 ℃ |
宿主菌: | NEB Stable |
拷贝数: | 高拷贝 |
插入基因信息 |
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克隆方法: | Gibson 克隆 |
5’测序引物 | GCTTCTTGTGTATATCCGGTC |
文献引用方法 |
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材料和方法部分: | pLS-SceI was a gift from Nadav Ahituv (Addgene plasmid # 137725 ; http://n2t.net/addgene:137725 ; RRID:Addgene_137725) |
买家导航
pLS-SceI载体 pLSSceI pLScel质粒基本信息
pLS-SceI质粒图谱 pLSSceI pLScel载体图谱和pLS-SceI载体序列 pLSSceI pLScel质粒序列多克隆位点信息
pLS-SceI质粒 pLSSceI pLScel载体简介
pLS-SceI质粒序列 pLSSceI pLScel载体序列
LOCUS pLS-SceI 7810 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Lentivirus reporter assay plasmid that contains a I-SceI site and a EGFP reporter gene.. ACCESSION . VERSION . KEYWORDS pLS-SceI SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7810) TITLE JOURNAL Unpublished REFERENCE 2 (bases 1 to 7810) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7810 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 1..380 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 382..580 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 535..555 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" LTR 598..778 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 825..950 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 1447..1680 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 1865..1909 /codon_start=1 /product="antigenic peptide corresponding to amino acids 655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik et al., 2013)" /label=gp41 peptide /note="recognized by the 2H10 single-chain llama nanobody" /translation="KNEQELLELDKWASL" misc_feature 2207..2324 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" regulatory 2828..2837 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 2834..3550 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(2879..2900) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(3140..3159) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 3487..3508 /label=EGFP-C /note="EGFP, forward primer" protein_bind 3573..3606 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." misc_feature 3662..4250 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(3715..3735) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(4133..4144) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" primer_bind complement(4253..4269) /label=KS primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4254..4270) /label=pBluescriptKS /note="For pBluescript vector" LTR 5579..5759 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" rep_origin complement(5821..6409) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5901..5920) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6580..7440) /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 7203..7222 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(7441..7545) /gene="bla" /label=AmpR promoter primer_bind complement(7620..7639) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" ORIGIN 1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc 61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca 121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga 181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc 241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct 301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat 361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc 421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt 481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa 541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag cgcgttttgc ctgtactggg 601 tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg 661 cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt 721 gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt 781 ggcgcccgaa cagggacttg aaagcgaaag ggaaaccaga ggagctctct cgacgcagga 841 ctcggcttgc tgaagcgcgc acggcaagag gcgaggggcg gcgactggtg agtacgccaa 901 aaattttgac tagcggaggc tagaaggaga gagatgggtg cgagagcgtc agtattaagc 961 gggggagaat tagatcgcga tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat 1021 ataaattaaa acatatagta tgggcaagca gggagctaga acgattcgca gttaatcctg 1081 gcctgttaga aacatcagaa ggctgtagac aaatactggg acagctacaa ccatcccttc 1141 agacaggatc agaagaactt agatcattat ataatacagt agcaaccctc tattgtgtgc 1201 atcaaaggat agagataaaa gacaccaagg aagctttaga caagatagag gaagagcaaa 1261 acaaaagtaa gaccaccgca cagcaagcgg ccggccgctg atcttcagac ctggaggagg 1321 agatatgagg gacaattgga gaagtgaatt atataaatat aaagtagtaa aaattgaacc 1381 attaggagta gcacccacca aggcaaagag aagagtggtg cagagagaaa aaagagcagt 1441 gggaatagga gctttgttcc ttgggttctt gggagcagca ggaagcacta tgggcgcagc 1501 gtcaatgacg ctgacggtac aggccagaca attattgtct ggtatagtgc agcagcagaa 1561 caatttgctg agggctattg aggcgcaaca gcatctgttg caactcacag tctggggcat 1621 caagcagctc caggcaagaa tcctggctgt ggaaagatac ctaaaggatc aacagctcct 1681 ggggatttgg ggttgctctg gaaaactcat ttgcaccact gctgtgcctt ggaatgctag 1741 ttggagtaat aaatctctgg aacagatttg gaatcacacg acctggatgg agtgggacag 1801 agaaattaac aattacacaa gcttaataca ctccttaatt gaagaatcgc aaaaccagca 1861 agaaaagaat gaacaagaat tattggaatt agataaatgg gcaagtttgt ggaattggtt 1921 taacataaca aattggctgt ggtatataaa attattcata atgatagtag gaggcttggt 1981 aggtttaaga atagtttttg ctgtactttc tatagtgaat agagttaggc agggatattc 2041 accattatcg tttcagaccc acctcccaac cccgagggga cccgacaggc ccgaaggaat 2101 agaagaagaa ggtggagaga gagacagaga cagatccatt cgattagtga acggatcggc 2161 actgcgtgcg ccaattctgc agacaaatgg cagtattcat ccacaatttt aaaagaaaag 2221 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac 2281 aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg 2341 acagcagaga tccagtttgg ttagtaccgg gcccggtgct ttgctctgag ccagcccacc 2401 agtttggaat gactcctttt tatgacttga attttcaagt ataaagtcta gtgctaaatt 2461 taatttgaac aactgtatag tttttgctgg ttgggggaag gaaaaaaaat ggtggcagtg 2521 tttttttcag aattagaagt gaaatgaaaa cttgttgtgt gtgaggattt ctaatgacat 2581 gtggtggttg catactgagt gaagccggtg agcattctgc catgtcaccc cctcgtgctc 2641 agtaatgtac tttacagaaa tcctaaactc aaaagattga tataaaccat gcttcttgtg 2701 tatatccggt ctcttctctg ggtagtctca ctcagcctgc atttctgcca gggcccgctc 2761 tagacctgca gggcatctaa tgccacttag ggataacagg gtaatgctat ccaggttaac 2821 accggtcgcc accatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct 2881 ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg 2941 cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt 3001 gccctggccc accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc 3061 cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga 3121 gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga 3181 gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa 3241 catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga 3301 caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag 3361 cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct 3421 gcccgacaac cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg 3481 cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga 3541 gctgtacaag taggaattcg tcgagggacc taataacttc gtatagcata cattatacga 3601 agttatacat gtttaagggt tccggttcca ctaggtacaa ttcgatatca agcttatcga 3661 taatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc 3721 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg 3781 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt 3841 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac 3901 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc 3961 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct 4021 gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct 4081 cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct 4141 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct 4201 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc atcgataccg 4261 tcgacctcga tcgagaccta gaaaaacatg gagcaatcac aagtagcaat acagcagcta 4321 ccaatgctga ttgtgcctgg ctagaagcac aagaggagga ggaggtgggt tttccagtca 4381 cacctcaggt accaagcatg gggtaaagta ctgttctcat cacatcatat caaggttata 4441 taccatcaat attgccacag atgttactta gccttttaat atttctctaa tttagtgtat 4501 atgcaatgat agttctctga tttctgagat tgagtttctc atgtgtaatg attatttaga 4561 gtttctcttt catctgttca aatttttgtc tagttttatt ttttactgat ttgtaagact 4621 tctttttata atctgcatat tacaattctc tttactgggg tgttgcaaat attttctgtc 4681 attctatggc ctgacttttc ttaatggttt tttaatttta aaaataagtc ttaatattca 4741 tgcaatctaa ttaacaatct tttctttgtg gttaggactt tgagtcataa gaaatttttc 4801 tctacactga agtcatgatg gcatgcttct atattatttt ctaaaagatt taaagttttg 4861 ccttctccat ttagacttat aattcactgg aatttttttg tgtgtatggt atgacatatg 4921 ggttcccttt tattttttac atataaatat atttccctgt ttttctaaaa aagaaaaaga 4981 tcatcatttt cccattgtaa aatgccatat ttttttcata ggtcacttac atatatcaat 5041 gggtctgttt ctgagctcta ctctatttta tcagcctcac tgtctatccc cacacatctc 5101 atgctttgct ctaaatcttg atatttagtg gaacattctt tcccattttg ttctacaaga 5161 atatttttgt tattgtcttt tgggcttcta tatacatttt agaatgaggt tggcaagttc 5221 tgtagatctt agccactttt taaaagaaaa ggggggactg gaagggctaa ttcactccca 5281 acgaagacaa gatatccttg atctgtggat ctaccacaca caaggctact tccctgattg 5341 gcagaactac acaccagggc cagggatcag atatccactg acctttggat ggtgctacaa 5401 gctagtacca gttgagcaag agaaggtaga agaagccaat gaaggagaga acacccgctt 5461 gttacaccct gtgagcctgc atgggatgga tgacccggag agagaagtat tagagtggag 5521 gtttgacagc cgcctagcat ttcatcacat ggcccgagag ctgcatccgg actgtactgg 5581 gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact 5641 gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg 5701 tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag 5761 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt 5821 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg 5881 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg 5941 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag 6001 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc 6061 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa 6121 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg 6181 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc 6241 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac 6301 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg 6361 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt 6421 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt 6481 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa 6541 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga 6601 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt 6661 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg 6721 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga 6781 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga 6841 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg 6901 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc 6961 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc 7021 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca 7081 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac 7141 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg 7201 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc 7261 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg 7321 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac 7381 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat 7441 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata 7501 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa 7561 agtgccacct gacgtcgacg gatcgggaga tctcccgatc ccctatggtg cactctcagt 7621 acaatctgct ctgatgccgc atagttaagc cagtatctgc tccctgcttg tgtgttggag 7681 gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt gaccgacaat 7741 tgcatgaaga atctgcttag ggttaggcgt tttgcgctgc ttcgcgatgt acgggccaga 7801 tatacgcgtt //
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pLS-SceI质粒 pLSSceI pLScel载体应用举例
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