质粒名称: | pJ241-FLEX |
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出品公司: | Addgene |
目录编号: | 18925 |
存储实验室: | Scott Sternson |
载体骨架基本信息 |
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载体名称: | pJ241 |
载体出品公司: | DNA2.0 |
空载体大小: | 2791 bp |
完整质粒大小: | 3063 bp |
载体类型: | Cre/Lox ; subcloning |
载体抗性: | Kanamycin |
生长温度: | 37 ℃ |
宿主菌: | DH5a |
拷贝数: | 高拷贝 |
插入基因信息 |
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插入基因名称: | FLEX switch / flip-excision switch |
物种来源: | 光合细菌 Rhodopseudomonas palustris |
插入大小: | 274 bp |
基因突变: | An internal Spe1 site is used for subcloning a gene into the FLEX switch. |
GenBank编号 | EU784738 |
启动子: | CMV |
克隆方法: | 限制性内切酶 |
5’克隆位点: | BamH1 (not destroyed) |
3’克隆位点: | BstB1, EcoR1, Pme1, Xba1 (not destroyed) |
5’测序引物 | n/a |
文献引用方法 |
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材料和方法部分: |
pJ241-FLEX was a gift from Scott Sternson (Addgene plasmid # 18925 ; http://n2t.net/addgene:18925 ; RRID:Addgene_18925)
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参考文献部分: | A FLEX switch targets Channelrhodopsin-2 to multiple cell types for imaging and long-range circuit mapping. Atasoy D, Aponte Y, Su HH, Sternson SM. J Neurosci. 2008 Jul 9. 28(28):7025-30. 10.1523/JNEUROSCI.1954-08.2008 PubMed 18614669 |
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LOCUS pJ241-FLEX 3063 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pJ241-FLEX SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3063) AUTHORS Atasoy D, Aponte Y, Su HH, Sternson SM TITLE A FLEX switch targets Channelrhodopsin-2 to multiple cell types for imaging and long-range circuit mapping. JOURNAL J Neurosci. 2008 Jul 9. 28(28):7025-30. PUBMED 18614669 REFERENCE 2 (bases 1 to 3063) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.biofeng.com FEATURES Location/Qualifiers source 1..3063 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 88..105 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 89..105 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" protein_bind 575..608 /label=FRT (minimal) /bound_moiety="FLP recombinase from the Saccharomyces cerevisiae 2u plasmid" /note="supports FLP-mediated excision but not integration (Turan and Bode, 2011)" protein_bind complement(655..688) /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." protein_bind complement(759..792) /label=lox2272 /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GGATACTT). lox2272 sites are compatible with each other, but incompatible with loxP or loxN sites (Lee and Saito, 1988)." protein_bind 799..832 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." protein_bind 895..928 /label=lox2272 /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GGATACTT). lox2272 sites are compatible with each other, but incompatible with loxP or loxN sites (Lee and Saito, 1988)." protein_bind complement(945..978) /label=FRT (minimal) /bound_moiety="FLP recombinase from the Saccharomyces cerevisiae 2u plasmid" /note="supports FLP-mediated excision but not integration (Turan and Bode, 2011)" primer_bind complement(1185..1204) /label=pENTR-R /note="pENTR vectors, reverse primer" promoter 1271..1362 /gene="bla" /label=AmpR promoter CDS 1363..2172 /codon_start=1 /gene="aph(3')-Ia" /product="aminoglycoside phosphotransferase" /label=KanR /note="confers resistance to kanamycin in bacteria or G418 (Geneticin(R)) in eukaryotes" /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF" rep_origin 2294..2882 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 2783..2802 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 3036..3053 /label=L4440 /note="L4440 vector, forward primer" ORIGIN 1 gcgagagtag ggaactgcca ggcatcaaac taagcagaag gcccctgacg gatggccttt 61 ttgcgtttct acaaactctt tctgtgttgt aaaacgacgg ccagtcttaa gctcgggccc 121 cctgggcggt tctgataacg agtaatcgtt aatccgcaaa taacgtaaaa acccgcttcg 181 gcgggttttt ttatgggggg agtttaggga aagagcattt gtcagaatat ttaagggcgc 241 ctgtcacttt gcttgatata tgagaattat ttaaccttat aaatgagaaa aaagcaacgc 301 actttaaata agatacgttg ctttttcgat tgatgaacac ctataattaa actattcatc 361 tattatttat gattttttgt atatacaata tttctagttt gttaaagaga attaagaaaa 421 taaatctcga aaataataaa gggaaaatca gtttttgata tcaaaattat acatgtcaac 481 gataatacaa aatataatac aaactataag atgttatcag tatttattat gcatttagaa 541 taaattttgt gtcgccctta ttcgactcac tatagaagtt cctattctct agaaagtata 601 ggaacttcac ttcattttct agagtttaaa cgaattcgaa cctcttcgag ggacataact 661 tcgtataatg tatgctatac gaagttatgg gtcgatggtg agatctggac tagagggtcg 721 atggtgatgc ttggatcaaa ataggaagaa tcccaatgat aacttcgtat aaagtatcct 781 atacgaagtt atactagtat aacttcgtat agcatacatt atacgaagtt atgggtcgat 841 ggtgatgctt ggcaattcgg gtcgatggtg aagcattggt cttcctattt tgatataact 901 tcgtatagga tactttatac gaagttatgg ggatccccac ttcagaagtt cctatacttt 961 ctagagaata ggaacttcac tatagagtcg aataagggcg acacccccta attagcccgg 1021 gcgaaaggcc cagtctttcg actgagcctt tcgttttatt tgatgcctgg cagttcccta 1081 ctctcgcatg gggagtcccc acactaccat cggcgctacg gcgtttcact tctgagttcg 1141 gcatggggtc aggtgggacc accgcgctac tgccgccagg caaacaaggg gtgttatgag 1201 ccatattcag gtataaatgg gctcgcgata atgttcagaa ttggttaatt ggttgtaaca 1261 ctgaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa 1321 taaccctgat aaatgcttca ataatattga aaaaggaaga atatgagcca tattcaacgg 1381 gaaacgtcga ggccgcgatt aaattccaac atggatgctg atttatatgg gtataaatgg 1441 gctcgcgata atgtcgggca atcaggtgcg acaatctatc gcttgtatgg gaagcccgat 1501 gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg ccaatgatgt tacagatgag 1561 atggtcagac taaactggct gacggaattt atgccacttc cgaccatcaa gcattttatc 1621 cgtactcctg atgatgcatg gttactcacc actgcgatcc ccggaaaaac agcgttccag 1681 gtattagaag aatatcctga ttcaggtgaa aatattgttg atgcgctggc agtgttcctg 1741 cgccggttgc actcgattcc tgtttgtaat tgtcctttta acagcgatcg cgtatttcgc 1801 ctcgctcagg cgcaatcacg aatgaataac ggtttggttg atgcgagtga ttttgatgac 1861 gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa tgcataaact tttgccattc 1921 tcaccggatt cagtcgtcac tcatggtgat ttctcacttg ataaccttat ttttgacgag 1981 gggaaattaa taggttgtat tgatgttgga cgagtcggaa tcgcagaccg ataccaggat 2041 cttgccatcc tatggaactg cctcggtgag ttttctcctt cattacagaa acggcttttt 2101 caaaaatatg gtattgataa tcctgatatg aataaattgc aatttcattt gatgctcgat 2161 gagtttttct aagcagagca ttacgctgac ttgacgggac ggcgcaagct catgaccaaa 2221 atcccttaac gtgagttacg cgcgcgtcgt tccactgagc gtcagacccc gtagaaaaga 2281 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa 2341 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga 2401 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt 2461 tagcccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt 2521 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat 2581 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct 2641 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca 2701 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag 2761 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc 2821 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga 2881 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca 2941 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag 3001 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg 3061 aag //
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