质粒名称: | ePop |
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出品公司: | Addgene |
目录编号: | 50953 |
存储实验室: | Lingchong You |
载体骨架基本信息 |
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载体名称: | pProTet.E |
载体出品公司: | Clontech |
空载体大小: | -- |
完整质粒大小: | 4196 bp |
载体修饰: | 将具有NdeI和NheI位点的40bp寡聚体引入pProTet.E的AatII位点 |
载体类型: | -- |
筛选标记: | -- |
载体抗性: | 氯霉素,25μg/mL |
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1: | E lysis protein |
物种来源1: | phage φX174 |
插入大小1: | 276 bp |
插入基因名称2: | LuxI |
物种来源2: | V. fischeri |
插入大小2: | 582 bp |
基因突变2: | mutated luxR |
GenBank编号2 | AY275714.1 |
插入基因名称3: | LuxR |
物种来源3: | Aliivibrio fischeri |
插入大小3: | 753 bp |
基因突变3: | 突变luxR中引入的早期终止密码子导致luxR功能的丧失;该基因的调控不是研究中观察到的振荡行为所必需的 |
文献引用方法 |
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材料和方法部分: | ePop was a gift from Lingchong You (Addgene plasmid # 50953 ; http://n2t.net/addgene:50953 ; RRID:Addgene_50953) |
参考文献部分: | Oscillations by minimal bacterial suicide circuits reveal hidden facets of host-circuit physiology. Marguet P, Tanouchi Y, Spitz E, Smith C, You L. PLoS One. 2010 Jul 30;5(7):e11909. doi: 10.1371/journal.pone.0011909. 10.1371/journal.pone.0011909 PubMed 20689598 |
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LOCUS ePop 4196 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expresses E lysis protein that causes in subpopulation death when threshold cell density is reached, resulting in oscillations. ACCESSION . VERSION . KEYWORDS ePop SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4196) AUTHORS Marguet P, Tanouchi Y, Spitz E, Smith C, You L TITLE Oscillations by minimal bacterial suicide circuits reveal hidden facets of host-circuit physiology. JOURNAL PLoS One. 2010 Jul 30;5(7):e11909. doi: 10.1371/journal.pone.0011909. PUBMED 20689598 REFERENCE 2 (bases 1 to 4196) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.biofeng.com FEATURES Location/Qualifiers source 1..4196 /organism="synthetic DNA construct" /mol_type="other DNA" protein_bind 443..459 /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)." primer_bind 448..470 /label=M13/pUC Reverse /note="In lacZ gene" CDS 661..690 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" protein_bind 867..886 /label=lac operator (symmetric) /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). The symmetric lac operator was optimized for tight binding of lac repressor." protein_bind 900..916 /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)." RBS 935..946 /note="strong bacterial ribosome binding site (Elowitz and Leibler, 2000)" terminator 2370..2456 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" rep_origin complement(2620..3208) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(2700..2719) /label=pBR322ori-F /note="pBR322 origin, forward primer" terminator 3296..3390 /label=lambda t0 terminator /note="transcription terminator from phage lambda" CDS complement(3411..4070) /codon_start=1 /gene="cat" /product="chloramphenicol acetyltransferase" /label=CmR /note="confers resistance to chloramphenicol" /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA" primer_bind 3985..4004 /label=CAT-R /note="Chloramphenicol resistance gene, reverse primer" promoter complement(4071..4173) /label=cat promoter /note="promoter of the E. coli cat gene encoding chloramphenicol acetyltransferase" ORIGIN 1 gacgtcagtc ctttgattct aataaattgg atttttgtca cactattgta tcgctgggaa 61 tacaattact taacataagc acctgtagga tcgtacaggt ttacgcaaga aaatggtttg 121 ttatagtcga ataaacgcaa gggaggttgg tatggtacgc tggactttgt gggataccct 181 cgctttcctg ctcctgttga gtttattgct gccgtcattg cttattatgt tcatcccgtc 241 aacattcaaa cggcctgtct catcatggaa ggcgctgaat ttacggaaaa cattattaat 301 ggcgtcgagc gtccggttaa agccgctgaa ttgttcgcgt ttaccttgcg tgtacgcgca 361 ggaaacactg acgttcttac tgacgcagaa gaaaacgtgc gtcaaaaatt acgtgcggaa 421 ggagtgagac gtctgtgtgg aattgtgagc ggataacaat ttcacacagg gccctcggac 481 accgaggaga atgtcaagag gcgaacacac aacgtcttgg agcgccagag gaggaacgag 541 ctaaaacgga gcttttttgc cctgcgtgac cagatcccgg agttggaaaa caatgaaaag 601 gcccccaagg tagttatcct taaaaaagcc acagcataca tcctgtccgt ccaagcagag 661 gagcaaaagc tcatttctga agaggacttg ttgcggaaac gacgagaaca gttgaaacac 721 aaacttgaac agctacggaa ctcttgtgcg taaggaaaag taaggaaaac gattccttct 781 aacagaaatg tcctgagcaa tcacctatga actgtcgact cgagcatagc atttttatcc 841 ataagattag cggatctaac ctttacaatt gtgagcgctc acaattatga tagattcaat 901 tgtgagcgga taacaatttc acacagaatt cattaaagag gagaaaggta cccatgaaaa 961 acataaatgc cgacgacaca tacagaataa ttaataaaat taaagcttgt agaagcaata 1021 atgatattaa tcaatgctta tctgatatga ctaaaatggt acattgtgaa tattatttac 1081 tcgcgatcat ttatcctcat tctatggtta aatctgatat ttcaattcta gataattacc 1141 ctaaaaaatg gaggcaatat tatgatgacg ctaatttaat aaaatatgat cctatagtag 1201 attattctaa ctccaatcat tcaccaatta attggaatat atttgaaaac aatgctgtaa 1261 ataaaaaatc tccaaatgta attaaagaag cgaaaacatc aggtcttatc actgggttta 1321 gtttccctat tcatacggct aacaatggct tcggaatgct tagttttgca cattcagaaa 1381 aagacaacta tatagatagt ttatttttac atgcgtgtat gaacatacca ttaattgttc 1441 cttctctagt tgataattat cgaaaaataa atatagcaaa taataaatca aacaacgatt 1501 taaccaaaag agaaaaagaa tgtttagcgt gggcatgcga aggaaaaagc tcttgggata 1561 tttcaaaaat attaggctgc agtgagcgta ctgtcacttt ccatttaacc aatgcgcaaa 1621 tgaaactcaa tacaacaaac cgctgccaaa gtatttctaa agcaatttta acaggagcaa 1681 ttgattgccc atactttaaa aattaagtac gggatccccg aataaacgca agggaggttg 1741 gtatgactat aatgataaaa aaatcggatt ttttggcaat tccatcggag gagtataaag 1801 gtattctaag tcttcgttat caagtgttta agcaaagact tgagtgggac ttagttgtag 1861 aaaataacct tgaatcagat gagtatgata actcaaatgc agaatatatt tatgcttgtg 1921 atgatactga aaatgtaagt ggatgctggc gtttattacc tacaacaggt gattatatgc 1981 tgaaaagtgt ttttcctgaa ttgcttggtc aacagagtgc tcccaaagat cctaatatag 2041 tcgaattaag tcgttttgct gtaggtaaaa atagctcaaa gataaataac tctgctagtg 2101 aaattacaat gaaactattt gaagctatat ataaacacgc tgttagtcaa ggtattacag 2161 aatatgtaac agtaacatca acagcaatag agcgattttt aaagcgtatt aaagttcctt 2221 gtcatcgtat tggagacaaa gaaattcatg tattaggtga tactaaatcg gttgtattgt 2281 ctatgcctat taatgaacag tttaaaaaag cagtcttaaa ttaatgttgt taagcttcgc 2341 ggccgcttaa ttaattaatc tagaggcatc aaataaaacg aaaggctcag tcgaaagact 2401 gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct cctgagtagg acaaatccgc 2461 cgccctagac ctaggcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat 2521 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca 2581 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc 2641 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata 2701 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc 2761 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc 2821 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga 2881 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc 2941 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag 3001 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag 3061 gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag 3121 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca 3181 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga 3241 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgactagtg cttggattct 3301 caccaataaa aaacgcccgg cggcaaccga gcgttctgaa caaatccaga tggagttctg 3361 aggtcattac tggatctatc aacaggagtc caagcgagct cgatatcaaa ttacgccccg 3421 ccctgccact catcgcagta ctgttgtaat tcattaagca ttctgccgac atggaagcca 3481 tcacagacgg catgatgaac ctgaatcgcc agcggcatca gcaccttgtc gccttgcgta 3541 taatatttgc ccatggtgaa aacgggggcg aagaagttgt ccatattggc cacgtttaaa 3601 tcaaaactgg tgaaactcac ccagggattg gctgagacga aaaacatatt ctcaataaac 3661 cctttaggga aataggccag gttttcaccg taacacgcca catcttgcga atatatgtgt 3721 agaaactgcc ggaaatcgtc gtggtattca ctccagagcg atgaaaacgt ttcagtttgc 3781 tcatggaaaa cggtgtaaca agggtgaaca ctatcccata tcaccagctc accgtctttc 3841 attgccatac gaaattccgg atgagcattc atcaggcggg caagaatgtg aataaaggcc 3901 ggataaaact tgtgcttatt tttctttacg gtctttaaaa aggccgtaat atccagctga 3961 acggtctggt tataggtaca ttgagcaact gactgaaatg cctcaaaatg ttctttacga 4021 tgccattggg atatatcaac ggtggtatat ccagtgattt ttttctccat tttagcttcc 4081 ttagctcctg aaaatctcga taactcaaaa aatacgcccg gtagtgatct tatttcatta 4141 tggtgaaagt tggaacctct tacgtgccga tcaacgtctc attttcgcca gatatc //
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