ePop

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相关技术服务:质粒构建    基因合成    质粒大提

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载体基本信息

质粒名称: ePop
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
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
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
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

载体图谱质粒图谱和多克隆位点信息

ePop质粒图谱

载体简介

载体序列

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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