质粒名称: | pK031.TRE-Cre (Supernova) |
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出品公司: | addgene |
目录编号: | 69136 |
存储实验室: | Takuji Iwasato |
载体骨架基本信息 |
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载体名称: | pTRE-tight |
载体出品公司: | Clontech |
空载体大小: | 2546bp |
完整质粒大小: | 3686 bp |
载体类型: | 哺乳细胞表达 |
载体抗性: | 氨苄霉素 |
生长温度: | 37 ℃ |
宿主菌: | DH5a |
拷贝数: | 未知 |
插入基因信息 |
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插入基因名称: | NLS-Cre |
物种来源: | SV40 , P1 phage |
克隆方法: | 未知 |
5’测序引物 | CEP_fwd_primer |
文献引用方法 |
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材料和方法部分: | pK031.TRE-Cre (Supernova) was a gift from Takuji Iwasato (Addgene plasmid # 69136 ; http://n2t.net/addgene:69136 ; RRID:Addgene_69136) |
参考文献部分: | NMDAR-regulated dynamics of layer 4 neuronal dendrites during thalamocortical reorganization in neonates. Mizuno H, Luo W, Tarusawa E, Saito YM, Sato T, Yoshimura Y, Itohara S, Iwasato T. Neuron. 2014 Apr 16;82(2):365-79. doi: 10.1016/j.neuron.2014.02.026. Epub 2014 Mar 27. 10.1016/j.neuron.2014.02.026 PubMe |
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载体基本信息
载体图谱质粒图谱和多克隆位点信息
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载体序列
LOCUS pK031.TRE-Cre (Supernova) 3686 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION For the Supernova, a series of vector systems for single-cell labeling and gene function sudies in vivo. ACCESSION . VERSION . KEYWORDS pK031.TRE-Cre (Supernova) SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3686) AUTHORS Mizuno H, Luo W, Tarusawa E, Saito YM, Sato T, Yoshimura Y, Itohara S, Iwasato T TITLE NMDAR-regulated dynamics of layer 4 neuronal dendrites during thalamocortical reorganization in neonates. JOURNAL Neuron. 2014 Apr 16;82(2):365-79. doi: 10.1016/j.neuron.2014.02.026. Epub 2014 Mar 27. PUBMED 24685175 REFERENCE 2 (bases 1 to 3686) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.biofeng.com FEATURES Location/Qualifiers source 1..3686 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 4..318 /label=tight TRE promoter /note="Tet-responsive promoter PTight, consisting of seven tet operator sequences followed by the minimal CMV promoter" protein_bind 12..30 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 48..66 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 83..101 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 119..137 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 155..173 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 190..208 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 226..244 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" primer_bind 291..315 /label=LNCX /note="Human CMV promoter, forward primer" CDS 395..415 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 419..1444 /codon_start=1 /product="site-specific recombinase" /label=Cre /note="Cre is a site-specific recombinase from bacteriophage P1. Recombination occurs at loxP sequences." /translation="NLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKMLLS VCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGLPR PSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAFLG IAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVERW ISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQRY LAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLEDG D" polyA_signal 1600..1681 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(1637..1656) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind complement(1692..1709) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(1863..2451) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(1943..1962) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(2622..3482) /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 3245..3264 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(3483..3587) /gene="bla" /label=AmpR promoter primer_bind 3655..3673 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" ORIGIN 1 ctcgagttta ctccctatca gtgatagaga acgtatgtcg agtttactcc ctatcagtga 61 tagagaacga tgtcgagttt actccctatc agtgatagag aacgtatgtc gagtttactc 121 cctatcagtg atagagaacg tatgtcgagt ttactcccta tcagtgatag agaacgtatg 181 tcgagtttat ccctatcagt gatagagaac gtatgtcgag tttactccct atcagtgata 241 gagaacgtat gtcgaggtag gcgtgtacgg tgggaggcct atataagcag agctcgttta 301 gtgaaccgtc agatcgcctg gagaattcgc aggatcagtc gacctgcagc ccaagctgta 361 ccccctcgaa actgacagga gaaccaccat gggcccaaag aagaagagaa aggtttcgaa 421 tttactgacc gtacaccaaa atttgcctgc attaccggtc gatgcaacga gtgatgaggt 481 tcgcaagaac ctgatggaca tgttcaggga tcgccaggcg ttttctgagc atacctggaa 541 aatgcttctg tccgtttgcc ggtcgtgggc ggcatggtgc aagttgaata accggaaatg 601 gtttcccgca gaacctgaag atgttcgcga ttatcttcta tatcttcagg cgcgcggtct 661 ggcagtaaaa actatccagc aacatttggg ccagctaaac atgcttcatc gtcggtccgg 721 gctgccacga ccaagtgaca gcaatgctgt ttcactggtt atgcggcgga tccgaaaaga 781 aaacgttgat gccggtgaac gtgcaaaaca ggctctagcg ttcgaacgca ctgatttcga 841 ccaggttcgt tcactcatgg aaaatagcga tcgctgccag gatatacgta atctggcatt 901 tctggggatt gcttataaca ccctgttacg tatagccgaa attgccagga tcagggttaa 961 agatatctca cgtactgacg gtgggagaat gttaatccat attggcagaa cgaaaacgct 1021 ggttagcacc gcaggtgtag agaaggcact tagcctgggg gtaactaaac tggtcgagcg 1081 atggatttcc gtctctggtg tagctgatga tccgaataac tacctgtttt gccgggtcag 1141 aaaaaatggt gttgccgcgc catctgccac cagccagcta tcaactcgcg ccctggaagg 1201 gatttttgaa gcaactcatc gattgattta cggcgctaag gatgactctg gtcagagata 1261 cctggcctgg tctggacaca gtgcccgtgt cggagccgcg cgagatatgg cccgcgctgg 1321 agtttcaata ccggagatca tgcaagctgg tggctggacc aatgtaaata ttgtcatgaa 1381 ctatatccgt aacctggata gtgaaacagg ggcaatggtg cgcctgctgg aagatggcga 1441 ttagccatta acgcgcggcc gcatcgataa gcttgtcgac gatatctcta gaggatcata 1501 atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc 1561 ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat 1621 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg 1681 cctcgagctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta 1741 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag 1801 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg 1861 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg 1921 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg 1981 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga 2041 agcgtggcgc tttctcaatg ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc 2101 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt 2161 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact 2221 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg 2281 cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt 2341 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt 2401 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct 2461 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg 2521 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt 2581 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt 2641 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc 2701 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg 2761 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc 2821 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg 2881 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca 2941 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga 3001 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct 3061 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg 3121 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca 3181 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata 3241 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct 3301 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact 3361 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa 3421 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc 3481 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga 3541 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga 3601 aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg 3661 cgtatcacga ggccctttcg tcttca //
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