pULTRA-CNF

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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: pULTRA-CNF
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
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
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
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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载体简介

载体序列

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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