质粒名称: | pTW051b4 |
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出品公司: | 爱迪基因 |
目录编号: | 116891 |
存储实验室: | David Liu |
载体骨架基本信息 |
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载体名称: | ColE1 |
载体出品公司: | -- |
载体抗性: | 大观霉素和链霉素 |
空载体大小: | -- |
完整质粒大小: | 6540 bp |
载体修饰: | -- |
载体类型: | -- |
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: | g3c |
物种来源: | Synthetic |
插入大小: | 1362 bp |
基因突变: | -- |
GenBank编号 | -- |
启动子 | -- |
标签/融合蛋白: | -- |
克隆方法: | -- |
5' 克隆位点: | -- |
3' 克隆位点: | -- |
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: | pTW051b4 was a gift from David Liu (Addgene plasmid # 116891 ; http://n2t.net/addgene:116891 ; RRID:Addgene_116891) |
参考文献部分: | Continuous directed evolution of proteins with improved soluble expression. Wang T, Badran AH, Huang TP, Liu DR. Nat Chem Biol. 2018 Aug 20. pii: 10.1038/s41589-018-0121-5. doi: 10.1038/s41589-018-0121-5. 10.1038/s41589-018-0121-5 [pii] PubMed 30127387 |
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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 pTW051b4 6540 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION medium stringency SE-PACE AP. ACCESSION . VERSION . KEYWORDS pTW051b4 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6540) AUTHORS Wang T, Badran AH, Huang TP, Liu DR TITLE Continuous directed evolution of proteins with improved soluble expression. JOURNAL Nat Chem Biol. 2018 Aug 20. pii: 10.1038/s41589-018-0121-5. doi: 10.1038/s41589-018-0121-5. PUBMED 30127387 REFERENCE 2 (bases 1 to 6540) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6540 /organism="synthetic DNA construct" /mol_type="other DNA" CDS complement(1..792) /codon_start=1 /gene="aadA" /product="aminoglycoside adenylyltransferase (Murphy, 1985)" /label=SmR /note="confers resistance to spectinomycin and streptomycin" /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK" promoter complement(793..864) /gene="bla" /label=AmpR promoter rep_origin 893..1481 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 1382..1401 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 1635..1652 /label=L4440 /note="L4440 vector, forward primer" misc_feature 1667..1807 /label=bom /note="basis of mobility region from pBR322" primer_bind complement(1733..1752) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 1893..1915 /label=pGEX 3' /note="pGEX vectors, reverse primer" CDS complement(1909..2100) /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" terminator 6120..6147 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" terminator 6239..6325 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" primer_bind 6478..6495 /label=pBAD Reverse /note="For vectors with E. coli araBAD promoter, reverse primer" ORIGIN 1 ttatttgccg actaccttgg tgatctcgcc tttcacgtag tggacaaatt cttccaactg 61 atctgcgcgc gaggccaagc gatcttcttc ttgtccaaga taagcctgtc tagcttcaag 121 tatgacgggc tgatactggg ccggcaggcg ctccattgcc cagtcggcag cgacatcctt 181 cggcgcgatt ttgccggtta ctgcgctgta ccaaatgcgg gacaacgtaa gcactacatt 241 tcgctcatct ccagcccagt cgggcggcga gttccatagc gttaaggttt catttagcgc 301 ctcaaataga tcctgttcag gaaccggatc aaagagttcc tccgccgctg gacctaccaa 361 ggcaacgcta tgttctcttg cttttgtcag caagatagcc agatcaatgt cgatcgtggc 421 tggctcgaag atacctgcaa gaatgtcatt gcgctgccat tctccaaatt gcagttcgcg 481 cttagctgga taacgccacg gaatgatgtc gtcgtgcaca acaatggtga cttctacagc 541 gcggagaatc tcgctctctc caggggaagc cgaagtttcc aaaaggtcgt tgatcaaagc 601 tcgccgcgtt gtttcatcaa gccttacggt caccgtaacc agcaaatcaa tatcactgtg 661 tggcttcagg ccgccatcca ctgcggagcc gtacaaatgt acggccagca acgtcggttc 721 gagatggcgc tcgatgacgc caactacctc tgatagttga gtcgatactt cggcgatcac 781 cgcttccctc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct 841 catgagcgga tacatatttg aatgtattta gaaaaatagg ccaaataggc cgttgagatc 901 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg 961 tttgtttgcc ggatcaagag ctaccacctc tttttccgaa ggtaactggc ttcagcagag 1021 cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact 1081 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg 1141 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc 1201 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg 1261 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg 1321 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag 1381 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc 1441 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct 1501 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc 1561 ctgattctgt ggataaccgt attacagcct ttgagtgagc tgataccgct cgccgcagcc 1621 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg atgcggtatt 1681 ttctccttac gcatctgtgc ggtatttcac accgcatatg gtgcactctc agtacaatct 1741 gctctgatgc cgcatagtta agccagtata cactccgcta tcgctacgtg actgggtcat 1801 ggctgcgccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtcttctccc 1861 ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc 1921 accgtcatca ccgaaacgcg cgaggcagct gcggtaaagc tcatcagcgt ggtcgtgaag 1981 cgattcacag atgtctgcct gttcatccgc gtccagctcg ttgagtttct ccagaagcgt 2041 taatgtctgg cttctgataa agcgggccat gttaagggcg gttttttcct gtttggtcac 2101 tgatgcctcc gtgtaagggg gatttctgtt catgggggta atgataccga tgaaacgaga 2161 gaggatgctc acgatacggg ttactgatga tgaacaagag gacatccggt caaataaaac 2221 gaaaggctca gtcgaaagac tgggcctttc gttttgctga ggagacttag ggaccctact 2281 aatacgactc cctataggga gaaagacctg caggtgcagt aaaaaaggaa aaaaatgatc 2341 aaaattgcca cgcgtaaata tttaggcaaa cagaatgttt atgatatcgg tgtcgagcgc 2401 gatcataatt tcgcgctgaa aaacggcttt atcgccagca attgttttaa tgttgttcct 2461 ttctattctc actccgctga aactgttgaa agttgtttag caaaacccca tacagaaaat 2521 tcatttacta acgtctggaa agacgacaaa actttagatc gttacgctaa ctatgagggc 2581 tgtctgtgga atgctacagg cgttgtagtt tgtactggtg acgaaactca gtgttacggt 2641 acatgggttc ctattgggct tgctatccct gaaaatgagg gtggtggctc tgagggtggc 2701 ggttctgagg gtggcggttc tgagggtggc ggtactaaac ctcctgagta cggtgataca 2761 cctattccgg gctatactta tatcaaccct ctcgacggca cttatccgcc tggtactgag 2821 caaaaccccg ctaatcctaa tccttctctt gaggagtctc agcctcttaa tactttcatg 2881 tttcagaata ataggttccg aaataggcag ggggcattaa ctgtttatac gggcactgtt 2941 actcaaggca ctgaccccgt taaaacttat taccagtaca ctcctgtatc atcaaaagcc 3001 atgtatgacg cttactggaa cggtaaattc agagactgcg ctttccattc tggctttaat 3061 gaggatccat tcgtttgtga atatcaaggc caatcgtctg acctgcctca acctcctgtc 3121 aatgctggcg gcggctctgg tggtggttct ggtggcggct ctgagggtgg tggctctgag 3181 ggtggcggtt ctgagggtgg cggctctgag ggaggcggtt ccggtggtgg ctctggttcc 3241 ggtgattttg attatgaaaa gatggcaaac gctaataagg gggctatgac cgaaaatgcc 3301 gatgaaaacg cgctacagtc tgacgctaaa ggcaaacttg attctgtcgc tactgattac 3361 ggtgctgcta tcgatggttt cattggtgac gtttccggcc ttgctaatgg taatggtgct 3421 actggtgatt ttgctggctc taattcccaa atggctcaag tcggtgacgg tgataattca 3481 cctttaatga ataatttccg tcaatattta ccttccctcc ctcaatcggt tgaatgtcgc 3541 ccttttgtct ttggcgctgg taaaccttac gagttcagta tcgactgcga taagatcaac 3601 ctgttccgcg gtgtctttgc gtttctttta tatgttgcca cctttatgta tgtattttct 3661 acgtttgcta acatactgcg taataaggag tcttaaacgg agccaatgta cgcaaaaaac 3721 cccgcttcag cggggttttt tcgccaaaaa aaaccccgcc ctgtcagggg cggggttttt 3781 ttttagggcc ctactgactg ttacgcgaac gcgaagtccg actctaagat gtcacggagg 3841 ttcaagttac ctttagccgg aagtgctggc attttgtcca attgagactc gtgcaactgg 3901 tcagcgaact ggtcgtagaa atcagccagt acatcacaag actcatatgt gtcaaccata 3961 gtttcgcgca ctgctttgaa caggttcgca gcgtcagccg gaatggtacc gaaggagtcg 4021 tgaatcagtg caaaagattc gattccgtac ttctcgtgtg cccacactac agtcttacga 4081 aggtggctac cgtcttggct gtgtacaaag ttaggagcga taccagactc ctgtttgtgt 4141 gcatcaatct cgctatcttt gttggtgtta atggtaggct gtaagcggaa ctgaccgagg 4201 aacatcaggt tcaagcgcgt ctgaataggc ttcttgtatt cctgccacac agggaaacca 4261 tcaggagtta cccaatgcac agcgcaacgc ttgcgaagaa tctctccagt cttcttatct 4321 ttgacctcag cagccagcag cttagcagca gacttaagcc agttcattgc ttcaaccgca 4381 gctaccaccg tcacgctcac agattcccaa atcagcttag ccatgtatcc agcagcctga 4441 ttcggctgag tgaacatcag acccttgccg gaatcaatag ctggctgaat ggtatcttcc 4501 agcacttgtt gacggaagcc gaactctttg gacccgtaag ccagcgtcat gactgaactc 4561 ttagtcacac tgcgagtaac accgtaagcc agccattgac cagccagtgc cttagtgccc 4621 agcttgactt tctcagagat ttcaccagtg ttctcatcgg tcacggtaac tacttcgtta 4681 tcggtcccat tgattgcgtc tgcttgtaga atctcgttga ctttcttagc aacaatcccg 4741 tagatgtcct gaacggtttc actaggaagc aagttaaccg cgcgaccacc tacctcatct 4801 cggagcatcg cggagaagtg ctggatgcca gagcaagacc cgtcaaacgc cagcggaagg 4861 gagcagttat agctcaggcc gtggtgctgt accccagcgt actcaaagca gaacgcaagg 4921 aagcagaacg gagaatcttg ctcagcccac caagtgttct ccagtggaga cttagcgcaa 4981 gccatgatgt tctcgtggtt ttcctcaatg aacttgatgc gctcagggaa cggaacctta 5041 tcgacacccg cacagtttgc accgtggatt ttcagccagt agtaaccttc cttaccgatt 5101 ggtttacctt tcgccagcgt aagcagtcct ttggtcatat cgttaccttg cgggttgaac 5161 attgacacag cgtaaacacg accgcgccag tccatgttgt aagggaacca gatggcctta 5221 tggttagcaa acttattggc ttgctcaagc atgaactcaa ggctgatacg gcgagacttg 5281 cgagccttgt ccttgcggta cacagcagcg gcagcacgtt tccacgcggt gagagcctca 5341 ggattcatgt cgatgtcttc cggtttcatc gggagttctt cacgctcaat cgcagggatg 5401 tcctcgaccg gacaatgctt ccacttggtg attacgttgg cgaccgctag gactttcttg 5461 ttgattttcc atgcggtgtt ttgcgcaatg ttaatcgctt tgtacacctc aggcatgtaa 5521 acgtcttcgt agcgcatcag tgctttctta ctgtgagtac gcaccagcgc cagaggacga 5581 cgaccgttag cccaatagcc accaccagta atgccagtcc acggcttagg aggaactacg 5641 caaggttgga acatcggaga gatgccagcc agcgcacctg cacgggttgc gatagcctca 5701 gcgtattcag gtgcgagttc gatagtctca gagtcttgac ctactacgcc agcattttgg 5761 cggtgtaagc taaccattcc ggttgactca atgagcatct cgatgcagcg tactcctaca 5821 tgaatagagt cttccttatg ccacgaagac cacgcctcgc caccgagtag acccttagag 5881 agcatgtcag cctcgacaac ttgcataaat gctttcattt ttttttcctc cttactctag 5941 taaaagttaa acaaaattat ttgtagaggg aaaccgttgt ggtctccctg aatatagata 6001 ccgagcctta tgcatgcccg taaagttatc cagcaaccac tcatagacct agggcagcag 6061 atagggacga cgtggtgtta gctgtgctct agaatcctca gcgagtttgt agaaacgcaa 6121 aaaggccatc cgtcaggatg gccttctgct taatttgatg cctggcagtt tatggcgggc 6181 gtcctgcccg ccaccctccg ggccgttgct tcgcaacgtt caaatccgct cccggcggat 6241 ttgtcctact caggagagcg ttcaccgaca aacaacagat aaaacgaaag gcccagtctt 6301 tcgactgagc ctttcgtttt atttgatgcc tggcagttcc ctactctcgc atggggagac 6361 cccacactac catcggcgct acggcgtttc acttctgagt tcggcatggg gtcaggtggg 6421 accaccgcgc tactgccgcc aggcaaattc tgttttatca gaccgcttct gcgttctgat 6481 ttaatctgta tcaggctgaa aatcttctct catccgccaa aacagccatg cccatggcgt //