质粒名称: | pTU2-AmyE-Pveg-eGFP |
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出品公司: | 爱迪基因 |
目录编号: | 112792 |
存储实验室: | Chris French |
载体骨架基本信息 |
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载体名称: | pSB1C3 |
载体出品公司: | iGEM |
载体抗性: | 氯霉素 |
空载体大小: | 3185 bp |
完整质粒大小: | -- |
载体修饰: | -- |
载体类型: | 细菌表达 |
筛选标记: | -- |
高拷贝/低拷贝: | 高拷贝 |
生长条件: | 37 ℃ |
插入基因信息 |
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插入基因名称: | Pveg-eGFP |
物种来源: | Synthetic |
插入大小: | 1400 bp |
基因突变: | -- |
GenBank编号 | -- |
启动子 | -- |
标签/融合蛋白: | -- |
克隆方法: | 限制性内切酶 |
5' 克隆位点: | BsmB1(销毁后未知) |
3' 克隆位点: | BsmB1(销毁后未知) |
5' 测序引物 | -- |
3' 测序引物 | -- |
文献引用方法 |
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材料和方法部分: | pTU2-AmyE-Pveg-eGFP was a gift from Chris French (Addgene plasmid # 112792 ; http://n2t.net/addgene:112792 ; RRID:Addgene_112792) |
参考文献部分: | BacilloFlex: A modular DNA assembly toolkit for Bacillus subtilis synthetic biology. Wicke N, Radford D, Verrone V, Wipat A, French CE. bioRxiv 185108 10.1101/185108 |
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载体基本信息
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载体简介
载体序列
LOCUS pTU2-AmyE-Pveg-eGFP 4978 bp ds-DNA circular SYN 14-AUG-2018 DEFINITION Expresses GFP under the control of Pveg promoter and it is integrated into AmyE locus. Used to test correct transformation. ACCESSION . VERSION . KEYWORDS pTU2-AmyE-Pveg-eGFP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4978) AUTHORS Wicke N, Radford D, Verrone V, Wipat A, French CE TITLE BacilloFlex: A modular DNA assembly toolkit for Bacillus subtilis synthetic biology JOURNAL bioRxiv REFERENCE 2 (bases 1 to 4978) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 14, 2018 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4978 /organism="synthetic DNA construct" /mol_type="other DNA" misc_feature 1..22 /label=BioBrick prefix /note="BioBrick prefix for parts that do not start with ""ATG""" CDS 678..1394 /codon_start=1 /product="Aequoria victoria green fluorescent protein" /label=GFP /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE FVTAAGITHGMDELYK" primer_bind complement(708..736) /label=GFP-R /note="GFP, reverse primer. Does NOT anneal to EGFP" primer_bind 1331..1351 /label=GFP-F /note="GFP, forward primer. Does NOT anneal to EGFP" terminator 1407..1478 /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" terminator 1494..1521 /label=T7Te terminator /note="phage T7 early transcription terminator" terminator 2952..3009 /label=his operon terminator /note="This putative transcriptin terminator from the E. coli his operon has a 2-bp deletion introduced during synthesis. Its efficiency has not been determined." primer_bind complement(3033..3050) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(3204..3792) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(3284..3303) /label=pBR322ori-F /note="pBR322 origin, forward primer" terminator 3975..4069 /label=lambda t0 terminator /note="transcription terminator from phage lambda" CDS complement(4090..4749) /codon_start=1 /gene="cat" /product="chloramphenicol acetyltransferase" /label=CmR /note="confers resistance to chloramphenicol" /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA" primer_bind 4664..4683 /label=CAT-R /note="Chloramphenicol resistance gene, reverse primer" primer_bind 4911..4929 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" terminator 4933..4976 /label=bacterial terminator /note="putative bacterial transcription terminator" ORIGIN 1 gaattcgcgg ccgcttctag aggtctcact atatcttttg caaaacgatt caaaacctct 61 ttactgccgt tattcgctgg atttttattg ctgtttcatt tggttctggc aggaccggcg 121 gctgcgagtg ctgaaacggc gaacaaatcg aatgagctta cagcaccgtc gatcaaaagc 181 ggaaccattc ttcatgcatg gaattggtcg ttcaatacgt taaaacacaa tatgaaggat 241 attcatgatg caggatatac agccattcag acatctccga ttaaccaagt aaaggaaggg 301 aatcaaggag ataaaagcat gtcgaactgg tactggctgt atcagccgac atcgtatcaa 361 attggcaacc gttacttagg tactgaacaa gaatttaaag aaatgtgtgc agccgctgaa 421 gaatatggca taaaggtcat tgttgacgcg gtcatcaatc ataccaccag tgattatgcc 481 gcgatttcca atgaggttaa gagtattcca aactggacac atggaaacac acaaattaaa 541 aactggtctt gccctataat tttgtcaaaa taattttatt gacaacgtct tattaacgtt 601 gatataattt aaattttatt tgacaaaaat gggctcgtgt tgtacaataa atgtgtacat 661 attaagagga ggagcatatg cgtaaaggag aagaactttt cactggagtt gtcccaattc 721 ttgttgaatt agatggtgat gttaatgggc acaaattttc tgtcagtgga gagggtgaag 781 gtgatgcaac atacggaaaa cttaccctta aatttatttg cactactgga aaactacctg 841 ttccatggcc aacacttgtc actactttcg gttatggtgt tcaatgcttt gcgagatacc 901 cagatcatat gaaacagcat gactttttca agagtgccat gcccgaaggt tatgtacagg 961 aaagaactat atttttcaaa gatgacggga actacaagac acgtgctgaa gtcaagtttg 1021 aaggtgatac ccttgttaat agaatcgagt taaaaggtat tgattttaaa gaagatggaa 1081 acattcttgg acacaaattg gaatacaact ataactcaca caatgtatac atcatggcag 1141 acaaacaaaa gaatggaatc aaagttaact tcaaaattag acacaacatt gaagatggaa 1201 gcgttcaact agcagaccat tatcaacaaa atactccaat tggcgatggc cctgtccttt 1261 taccagacaa ccattacctg tccacacaat ctgccctttc gaaagatccc aacgaaaaga 1321 gagatcacat ggtccttctt gagtttgtaa cagctgctgg gattacacat ggcatggatg 1381 aactatacaa ataatcgacc aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg 1441 cctttcgttt tatctgttgt ttgtcggtga acgctctcta ctagagtcac actggctcac 1501 cttcgggtgg gcctttctgc gtttatatgt tccggcaccc attagttcaa caaacgaaaa 1561 ttggataaag tgggatattt ttaaaatata tatttatgtt acaataatat tgacttttaa 1621 aaaaggattg attctaatga agaaagcaga caagtaagcc tcctaaattc actttagata 1681 aaaatttagg aggcatatca aatgaacttt aataaaattg atttagacaa ttggaagaga 1741 aaagagatat ttaatcatta tttgaaccaa caaacgactt ttagtataac cacagaaatt 1801 gatattagtg ttttataccg aaacataaaa caagaaggat ataaatttta ccctgcattt 1861 attttcttag tgacaagggt gataaactca aatacagctt ttagaactgg ttacaatagc 1921 gacggagagt taggttattg ggataagtta gagccacttt atacaatttt tgatggtgta 1981 tctaaaacat tctctggtat ttggactcct gtaaagaatg acttcaaaga gttttatgat 2041 ttataccttt ctgatgtaga gaaatataat ggttcgggga aattgtttcc caaaacacct 2101 atacctgaaa atgctttttc tctttctatt attccatgga cttcatttac tgggtttaac 2161 ttaaatatca ataataatag taattacctt ctacccatta ttacagcagg aaaattcatt 2221 aataaaggta attcaatata tttaccgcta tctttacagg tacatcattc tgtttgtgat 2281 ggttatcatg caggattgtt tatgaactct attcaggaat tgtcagatag gcctaatgac 2341 tggcttttat aatatgagat aatgccgact gtacttttta cagtcggaag ttaggctggg 2401 cggtgatagc ttctcgttca ggcagtacgc ctcttttctt ttccagacct gagggaggcg 2461 gaaatggtgt gaggttcccg gggaaaagcc aaataggcga tcgcgggagt gctttatttg 2521 aagatcaggc tatcactgcg gtcaatagat ttcacaatgt gatggctgga cagcctgagg 2581 aactctcgaa cccgaatgga aacaaccaga tatttatgaa tcagcgcggc tcacatggcg 2641 ttgtgctggc aaatgcaggt tcatcctctg tctctatcaa tacggcaaca aaattgcctg 2701 atggcaggta tgacaataaa gctggagcgg gttcatttca agtgaacgat ggtaaactga 2761 caggcacgat caatgccagg tctgtagctg tgctttatcc tgatgatatt gcaaaagcgc 2821 ctcatgtttt ccttgagaat tacaaaacag gtgtaacaca ttctttcaat gatcaactga 2881 cgattacctt gcgtgcagat gcgaatacaa caaaagccgt ttatcaaatc ttaggtacag 2941 agaccctgca gtccggcaaa aaagggcaag gtgtcaccac cctgcccttt ttctttaaaa 3001 ccgaaaagat tacttcgcgt tatgcaggct tcctcgctca ctgactcgct gcgctcggtc 3061 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa 3121 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt 3181 aaaaaggccg cgttgctggc gtttttccac aggctccgcc cccctgacga gcatcacaaa 3241 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt 3301 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg 3361 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc 3421 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc 3481 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta 3541 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct 3601 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc 3661 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa 3721 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa 3781 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa 3841 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt 3901 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac 3961 agctcgaggc ttggattctc accaataaaa aacgcccggc ggcaaccgag cgttctgaac 4021 aaatccagat ggagttctga ggtcattact ggatctatca acaggagtcc aagcgagctc 4081 gatatcaaat tacgccccgc cctgccactc atcgcagtac tgttgtaatt cattaagcat 4141 tctgccgaca tggaagccat cacaaacggc atgatgaacc tgaatcgcca gcggcatcag 4201 caccttgtcg ccttgcgtat aatatttgcc catggtgaaa acgggggcga agaagttgtc 4261 catattggcc acgtttaaat caaaactggt gaaactcacc cagggattgg ctgacacgaa 4321 aaacatattc tcaataaacc ctttagggaa ataggccagg ttttcaccgt aacacgccac 4381 atcttgcgaa tatatgtgta gaaactgccg gaaatcgtcg tggtattcac tccagagcga 4441 tgaaaacgtt tcagtttgct catggaaaac ggtgtaacaa gggtgaacac tatcccatat 4501 caccagctca ccgtctttca ttgccatacg aaattccgga tgagcattca tcaggcgggc 4561 aagaatgtga ataaaggccg gataaaactt gtgcttattt ttctttacgg tctttaaaaa 4621 ggccgtaata tccagctgaa cggtctggtt ataggtacat tgagcaactg actgaaatgc 4681 ctcaaaatgt tctttacgat gccattggga tatatcaacg gtggtatatc cagtgatttt 4741 tttctccatt ttagcttcct tagctcctga aaatctcgat aactcaaaaa atacgcccgg 4801 tagtgatctt atttcattat ggtgaaagtt ggaacctctt acgtgcccga tcaactcgag 4861 tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta 4921 tcacgaggca gaatttcaga taaaaaaaat ccttagcttt cgctaaggat gatttctg //
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