质粒名称: | pULTRA-CNF |
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出品公司: | addgene |
目录编号: | 48215 |
存储实验室: | Peter Schultz |
载体骨架基本信息 |
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载体名称: | pULTRA |
载体出品公司: | Schultz lab |
完整质粒大小: | 4977bp |
载体类型: | 细菌细胞表达 |
载体抗性: | 壮观霉素,Spectinomycin, 50 μg/mL |
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1: | polyspecific MJ tyrosyl synthetase |
物种来源: | M. jannaschii |
基因突变: | TY32L, L65V, F108W, Q109M, D158G, I159A |
启动子: | tacI |
标签/融合蛋白: | 人细胞色素c氧化酶亚基VIII(N端) |
克隆方法: | 限制性内切酶 |
5’克隆位点: | NotI(未破坏) |
3’克隆位点: | NotI (未破坏) |
5’测序引物: | Tac promoter |
插入基因名称2: | tRNA |
启动子: | proK |
克隆方法: | 限制性内切酶 |
5’克隆位点: | SbfI (unknown if destroyed) |
3’克隆位点: | NcoI (unknown if destroyed) |
5’测序引物: | na |
文献引用方法 |
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材料和方法部分: | pULTRA-CNF was a gift from Peter Schultz (Addgene plasmid # 48215 ; http://n2t.net/addgene:48215 ; RRID:Addgene_48215) |
参考文献部分: | A genetically encoded infrared probe. Schultz KC, Supekova L, Ryu Y, Xie J, Perera R, Schultz PG. J Am Chem Soc. 2006 Nov 1;128(43):13984-5. 10.1021/ja0636690 PubMed 17061854 |
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LOCUS pULTRA-CNF 4977 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION highly efficient suppressor plasmid. ACCESSION . VERSION . KEYWORDS pULTRA-CNF SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4977) AUTHORS Schultz KC, Supekova L, Ryu Y, Xie J, Perera R, Schultz PG TITLE A genetically encoded infrared probe. JOURNAL J Am Chem Soc. 2006 Nov 1;128(43):13984-5. PUBMED 17061854 REFERENCE 2 (bases 1 to 4977) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.biofeng.com FEATURES Location/Qualifiers source 1..4977 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 11..39 /label=tac promoter /note="strong E. coli promoter; hybrid between the trp and lac UV5 promoters" protein_bind 47..63 /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 complement(1068..1085) /label=pBAD Reverse /note="For vectors with E. coli araBAD promoter, reverse primer" regulatory 1709..1718 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS complement(1901..2692) /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(2693..2784) /gene="bla" /label=AmpR promoter rep_origin complement(2832..3570) /direction=LEFT /label=CloDF13 ori /note="Plasmids containing the CloDF13 (CDF) origin of replication can be propagated in E. coli cells that contain additional plasmids with compatible origins." CDS complement(3780..4862) /codon_start=1 /gene="lacI" /product="lac repressor" /label=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)." /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR ALADSLMQLARQVSRLESGQ" primer_bind 4824..4843 /label=LacI-R /note="LacI, reverse primer" promoter complement(4863..4940) /gene="lacI" /label=lacI promoter ORIGIN 1 tagggagctg ttgacaatta atcatcggct cgtataatgt gtggaattgt gagcggataa 61 caatttcaca aaggaggtgc ggccgcatgg acgaatttga aatgataaag agaaacacat 121 ctgaaattat cagcgaggaa gagttaagag aggttttaaa aaaagatgaa aaatctgctc 181 tgataggttt tgaaccaagt ggtaaaatac atttagggca ttatctccaa ataaaaaaga 241 tgattgattt acaaaatgct ggatttgata taattatagt tttggctgat ttacatgcct 301 atttaaacca gaaaggagag ttggatgaga ttagaaaaat aggagattat aacaaaaaag 361 tttttgaagc aatggggtta aaggcaaaat atgtttatgg aagtgaatgg atgcttgata 421 aggattatac actgaatgtc tatagattgg ctttaaaaac taccttaaaa agagcaagaa 481 ggagtatgga acttatagca agagaggatg aaaatccaaa ggttgctgaa gttatctatc 541 caataatgca ggttaatggt gctcattatc ttggcgttga tgttgcagtt ggggggatgg 601 agcagagaaa aatacacatg ttagcaaggg agcttttacc aaaaaaggtt gtttgtattc 661 acaaccctgt cttaacgggt ttggatggag aaggaaagat gagttcttca aaagggaatt 721 ttatagctgt tgatgactct ccagaagaga ttagggctaa gataaagaaa gcatactgcc 781 cagctggagt tgttgaagga aatccaataa tggagatagc taaatacttc cttgaatatc 841 ctttaaccat aaaaaggcca gaaaaatttg gtggagattt gacagttaat agctatgagg 901 agttagagag tttatttaaa aataaggaat tgcatccaat ggatttaaaa aatgctgtag 961 ctgaagaact tataaagatt ttagagccaa ttagaaagag attataagcg gccgcgttta 1021 aacggtctcc agcttggctg ttttggcgga tgagagaaga ttttcagcct gatacagatt 1081 aaatcagaac gcagaagcgg tctgataaaa cagaatttgc ctggcggcag tagcgcggtg 1141 gtcccacctg accccatgcc gaactcagaa gtgaaacgcc gtagcgccga tggtagtgtg 1201 gggtctcccc atgcgagagt agggaactgc caggcatcaa ataaaacgaa aggctcagtc 1261 gaaagactgg gccttgtttg tgagctcccg gtcatcaatc atccccataa tccttgttag 1321 cctgcaggta attccgcttc gcaacatgtg agcaccggtt tattgactac cggaagcagt 1381 gtgaccgtgt gcttctcaaa tgcctgaggc cagtttgctc aggctctccc cgtggaggta 1441 ataattgacg atatgatcag tgcacggcta actaagcggc ctgctgactt tctcgccgat 1501 caaaaggcat tttgctatta agggattgac gagggcgtat ctgcgcagta agatgcgccc 1561 cgcattccgg cggtagttca gcagggcaga acggcggact ctaaatccgc atggcagggg 1621 ttcaaatccc ctccgccgga ccaaattcga aaagcctgct caacgagcag gcttttttgc 1681 atgctcgagc agctcagggt cgaatttgcc atggcggcca ccaggtacca ccggcgcctc 1741 aggcatttga gaagcacacg gtcacactgc ttccggtagt caataaaccg gtaaaccagc 1801 aatagacata agcggctatt taacgaccct gccctgaacc gacgaccggg tcatcgtggc 1861 cggatcttgc ggcccctcgg cttgaacgaa ttgttagaca ttatttgccg actaccttgg 1921 tgatctcgcc tttcacgtag tggacaaatt cttccaactg atctgcgcgc gaggccaagc 1981 gatcttcttc ttgtccaaga taagcctgtc tagcttcaag tatgacgggc tgatactggg 2041 ccggcaggcg ctccattgcc cagtcggcag cgacatcctt cggcgcgatt ttgccggtta 2101 ctgcgctgta ccaaatgcgg gacaacgtaa gcactacatt tcgctcatcg ccagcccagt 2161 cgggcggcga gttccatagc gttaaggttt catttagcgc ctcaaataga tcctgttcag 2221 gaaccggatc aaagagttcc tccgccgctg gacctaccaa ggcaacgcta tgttctcttg 2281 cttttgtcag caagatagcc agatcaatgt cgatcgtggc tggctcgaag atacctgcaa 2341 gaatgtcatt gcgctgccat tctccaaatt gcagttcgcg cttagctgga taacgccacg 2401 gaatgatgtc gtcgtgcaca acaatggtga cttctacagc gcggagaatc tcgctctctc 2461 caggggaagc cgaagtttcc aaaaggtcgt tgatcaaagc tcgccgcgtt gtttcatcaa 2521 gccttacggt caccgtaacc agcaaatcaa tatcactgtg tggcttcagg ccgccatcca 2581 ctgcggagcc gtacaaatgt acggccagca acgtcggttc gagatggcgc tcgatgacgc 2641 caactacctc tgatagttga gtcgatactt cggcgatcac cgcttccctc atactcttcc 2701 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg 2761 aatgtattta gaaaaataaa caaatagcta gctcactcgg tcgctacgct ccgggcgtga 2821 gactgcggcg ggcgctgcgg acacatacaa agttacccac agattccgtg gataagcagg 2881 ggactaacat gtgaggcaaa acagcagggc cgcgccggtg gcgtttttcc ataggctccg 2941 ccctcctgcc agagttcaca taaacagacg cttttccggt gcatctgtgg gagccgtgag 3001 gctcaaccat gaatctgaca gtacgggcga aacccgacag gacttaaaga tccccaccgt 3061 ttccggcggg tcgctccctc ttgcgctctc ctgttccgac cctgccgttt accggatacc 3121 tgttccgcct ttctccctta cgggaagtgt ggcgctttct catagctcac acactggtat 3181 ctcggctcgg tgtaggtcgt tcgctccaag ctgggctgta agcaagaact ccccgttcag 3241 cccgactgct gcgccttatc cggtaactgt tcacttgagt ccaacccgga aaagcacggt 3301 aaaacgccac tggcagcagc cattggtaac tgggagttcg cagaggattt gtttagctaa 3361 acacgcggtt gctcttgaag tgtgcgccaa agtccggcta cactggaagg acagatttgg 3421 ttgctgtgct ctgcgaaagc cagttaccac ggttaagcag ttccccaact gacttaacct 3481 tcgatcaaac cacctcccca ggtggttttt tcgtttacag ggcaaaagat tacgcgcaga 3541 aaaaaaggat ctcaagaaga tcctttgatc ttttctactg aaccgctcta gatttcagtg 3601 caatttatct cttcaaatgt agcacctgaa gtcagcccca tacgatataa gttgtaattc 3661 tcatgttagt catgccccgc gcccaccgga aggagctgac tgggttgaag gctctcaagg 3721 gcatcggtcg agatcccggt gcctaatgag tgagctaact tacattaatt gcgttgcgct 3781 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac 3841 gcgcggggag aggcggtttg cgtattgggc gccagggtgg tttttctttt caccagtgag 3901 acgggcaaca gctgattgcc cttcaccgcc tggccctgag agagttgcag caagcggtcc 3961 acgctggttt gccccagcag gcgaaaatcc tgtttgatgg tggttaacgg cgggatataa 4021 catgagctgt cttcggtatc gtcgtatccc actaccgaga tgtccgcacc aacgcgcagc 4081 ccggactcgg taatggcgcg cattgcgccc agcgccatct gatcgttggc aaccagcatc 4141 gcagtgggaa cgatgccctc attcagcatt tgcatggttt gttgaaaacc ggacatggca 4201 ctccagtcgc cttcccgttc cgctatcggc tgaatttgat tgcgagtgag atatttatgc 4261 cagccagcca gacgcagacg cgccgagaca gaacttaatg ggcccgctaa cagcgcgatt 4321 tgctggtgac ccaatgcgac cagatgctcc acgcccagtc gcgtaccgtc ttcatgggag 4381 aaaataatac tgttgatggg tgtctggtca gagacatcaa gaaataacgc cggaacatta 4441 gtgcaggcag cttccacagc aatggcatcc tggtcatcca gcggatagtt aatgatcagc 4501 ccactgacgc gttgcgcgag aagattgtgc accgccgctt tacaggcttc gacgccgctt 4561 cgttctacca tcgacaccac cacgctggca cccagttgat cggcgcgaga tttaatcgcc 4621 gcgacaattt gcgacggcgc gtgcagggcc agactggagg tggcaacgcc aatcagcaac 4681 gactgtttgc ccgccagttg ttgtgccacg cggttgggaa tgtaattcag ctccgccatc 4741 gccgcttcca ctttttcccg cgttttcgca gaaacgtggc tggcctggtt caccacgcgg 4801 gaaacggtct gataagagac accggcatac tctgcgacat cgtataacgt tactggtttc 4861 acattcacca ccctgaattg actctcttcc gggcgctatc atgccatacc gcgaaaggtt 4921 ttgcgccatt cgatggtgtc cgggatctcg acgctctccc ttatgcgact cctgcat //
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