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載體描述

There are a range of Gluthatione Sepharose? prepacked column and bulk media products available to purify GST Fusion proteins. For manual purification of sample volumes up to 600 μl use GST SpinTrap? microspin columns or GST MultiTrap? 4B 96-well plates. For sample volumes between 600 μl and 10 ml use GST GraviTrap? gravity flow column. Where sample volumes are above 10 ml, use LabMate? reservoir together with GST GraviTrap. All formats described can be used for preparation of samples in parallel. In addition GST HiTrap? 1 and 5 ml columns and GST HiPrep? FF 16/10 column are available for purification in a chromatography system such as the ?KTA? design system. Alternatively, Gluthatione Sepharose bulk media are available from 10 ml up to 500 ml. A GST Bulk Kit is also available combining 10 ml Gluthatione Sepharose 4B bulk medium and 5 empty gravity flow columns with required buffers. For simplified buffer preparation use the GST Buffer Kit. Ordering information for all associated products is listed below.

載體序列

LOCUS       Exported                4984 bp ds-DNA     circular SYN 15-MAR-2019

DEFINITION  synthetic circular DNA

ACCESSION   .

VERSION     .

KEYWORDS    .

SOURCE      synthetic DNA construct

  ORGANISM  synthetic DNA construct

REFERENCE   1  (bases 1 to 4984)

  AUTHORS   .

  TITLE     Direct Submission

  JOURNAL   Exported Friday, Mar 15, 2019 from SnapGene Viewer 4.3.4

FEATURES             Location/Qualifiers

     source          1..4984

                     /organism="synthetic DNA construct"

                     /mol_type="other DNA"

     promoter        183..211

                     /label=tac promoter

                     /note="strong E. coli promoter; hybrid between the trp and

                     lac UV5 promoters"

     protein_bind    219..235

                     /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)."

     CDS             258..911

                     /codon_start=1

                     /product="glutathione S-transferase from Schistosoma

                     japonicum"

                     /label=GST

                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK

                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY

                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL

                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"

     primer_bind     869..891

                     /label=pGEX5'

     CDS             918..941

                     /codon_start=1

                     /product="recognition and cleavage site for human

                     rhinovirus 3C and PreScission proteases"

                     /label=HRV 3C site

                     /translation="LEVLFQGP"

     misc_feature    945..992

                     /label=MCS

                     /note="Multi clone site"

     primer_bind     complement(1034..1056)

                     /label=pGEX3'

     promoter        1287..1391

                     /gene="bla"

                     /label=AmpR promoter

     CDS             1392..2252

                     /codon_start=1

                     /gene="bla"

                     /product="beta-lactamase"

                     /label=AmpR

                     /note="confers resistance to ampicillin, carbenicillin, and

                     related antibiotics"

                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI

                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS

                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW

                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA

                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS

                     LIKHW"

     rep_origin      2423..3011

                     /direction=RIGHT

                     /label=ori

                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of

                     replication"

     promoter        3255..3332

                     /gene="lacI (mutant)"

                     /label=lacIq promoter

                     /note="In the lacIq allele, a single base change in the

                     promoter boosts expression of the lacI gene about 10-fold."

     CDS             3333..4415

                     /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

                     YIPPSTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR

                     ALADSLMQLARQVSRLESGQ"

     protein_bind    4428..4449

                     /label=CAP binding site

                     /bound_moiety="E. coli catabolite activator protein"

                     /note="CAP binding activates transcription in the presence

                     of cAMP."

     promoter        4464..4494

                     /label=lac promoter

                     /note="promoter for the E. coli lac operon"

     protein_bind    4502..4518

                     /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     4526..4542

                     /label=M13 rev

                     /note="common sequencing primer, one of multiple similar

                     variants"

     CDS             4538..4984

                     /codon_start=1

                     /gene="lacZ fragment"

                     /product="LacZ-alpha fragment of beta-galactosidase"

                     /label=lacZ-alpha

                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART

                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV

                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGI"

     primer_bind     complement(4558..4574)

                     /label=M13 fwd

                     /note="common sequencing primer, one of multiple similar

                     variants"

ORIGIN

        1 acgttatcga ctgcacggtg caccaatgct tctggcgtca ggcagccatc ggaagctgtg

       61 gtatggctgt gcaggtcgta aatcactgca taattcgtgt cgctcaaggc gcactcccgt

      121 tctggataat gttttttgcg ccgacatcat aacggttctg gcaaatattc tgaaatgagc

      181 tgttgacaat taatcatcgg ctcgtataat gtgtggaatt gtgagcggat aacaatttca

      241 cacaggaaac agtattcatg tcccctatac taggttattg gaaaattaag ggccttgtgc

      301 aacccactcg acttcttttg gaatatcttg aagaaaaata tgaagagcat ttgtatgagc

      361 gcgatgaagg tgataaatgg cgaaacaaaa agtttgaatt gggtttggag tttcccaatc

      421 ttccttatta tattgatggt gatgttaaat taacacagtc tatggccatc atacgttata

      481 tagctgacaa gcacaacatg ttgggtggtt gtccaaaaga gcgtgcagag atttcaatgc

      541 ttgaaggagc ggttttggat attagatacg gtgtttcgag aattgcatat agtaaagact

      601 ttgaaactct caaagttgat tttcttagca agctacctga aatgctgaaa atgttcgaag

      661 atcgtttatg tcataaaaca tatttaaatg gtgatcatgt aacccatcct gacttcatgt

      721 tgtatgacgc tcttgatgtt gttttataca tggacccaat gtgcctggat gcgttcccaa

      781 aattagtttg ttttaaaaaa cgtattgaag ctatcccaca aattgataag tacttgaaat

      841 ccagcaagta tatagcatgg cctttgcagg gctggcaagc cacgtttggt ggtggcgacc

      901 atcctccaaa atcggatctg gaagttctgt tccaggggcc cctgggatcc ccggaattcc

      961 cgggtcgact cgagcggccg catcgtgact gactgacgat ctgcctcgcg cgtttcggtg

     1021 atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag

     1081 cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg

     1141 gcgcagccat gacccagtca cgtagcgata gcggagtgta taattcttga agacgaaagg

     1201 gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt

     1261 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac

     1321 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa

     1381 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat

     1441 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc

     1501 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga

     1561 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg

     1621 cggtattatc ccgtgttgac gccgggcaag agcaactcgg tcgccgcata cactattctc

     1681 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag

     1741 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc

     1801 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg

     1861 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg

     1921 acaccacgat gcctgcagca atggcaacaa cgttgcgcaa actattaact ggcgaactac

     1981 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac

     2041 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg

     2101 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg

     2161 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg

     2221 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac

     2281 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg

     2341 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg

     2401 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc

     2461 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc

     2521 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt

     2581 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc

     2641 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact

     2701 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac

     2761 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag

     2821 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg

     2881 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg

     2941 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga

     3001 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt

     3061 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct

     3121 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg

     3181 aggaagcgga agagcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac

     3241 accgcataaa ttccgacacc atcgaatggt gcaaaacctt tcgcggtatg gcatgatagc

     3301 gcccggaaga gagtcaattc agggtggtga atgtgaaacc agtaacgtta tacgatgtcg

     3361 cagagtatgc cggtgtctct tatcagaccg tttcccgcgt ggtgaaccag gccagccacg

     3421 tttctgcgaa aacgcgggaa aaagtggaag cggcgatggc ggagctgaat tacattccca

     3481 accgcgtggc acaacaactg gcgggcaaac agtcgttgct gattggcgtt gccacctcca

     3541 gtctggccct gcacgcgccg tcgcaaattg tcgcggcgat taaatctcgc gccgatcaac

     3601 tgggtgccag cgtggtggtg tcgatggtag aacgaagcgg cgtcgaagcc tgtaaagcgg

     3661 cggtgcacaa tcttctcgcg caacgcgtca gtgggctgat cattaactat ccgctggatg

     3721 accaggatgc cattgctgtg gaagctgcct gcactaatgt tccggcgtta tttcttgatg

     3781 tctctgacca gacacccatc aacagtatta ttttctccca tgaagacggt acgcgactgg

     3841 gcgtggagca tctggtcgca ttgggtcacc agcaaatcgc gctgttagcg ggcccattaa

     3901 gttctgtctc ggcgcgtctg cgtctggctg gctggcataa atatctcact cgcaatcaaa

     3961 ttcagccgat agcggaacgg gaaggcgact ggagtgccat gtccggtttt caacaaacca

     4021 tgcaaatgct gaatgagggc atcgttccca ctgcgatgct ggttgccaac gatcagatgg

     4081 cgctgggcgc aatgcgcgcc attaccgagt ccgggctgcg cgttggtgcg gatatctcgg

     4141 tagtgggata cgacgatacc gaagacagct catgttatat cccgccgtca accaccatca

     4201 aacaggattt tcgcctgctg gggcaaacca gcgtggaccg cttgctgcaa ctctctcagg

     4261 gccaggcggt gaagggcaat cagctgttgc ccgtctcact ggtgaaaaga aaaaccaccc

     4321 tggcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg

     4381 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag

     4441 ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga

     4501 attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta cggattcact

     4561 ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct

     4621 tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc

     4681 ttcccaacag ttgcgcagcc tgaatggcga atggcgcttt gcctggtttc cggcaccaga

     4741 agcggtgccg gaaagctggc tggagtgcga tcttcctgag gccgatactg tcgtcgtccc

     4801 ctcaaactgg cagatgcacg gttacgatgc gcccatctac accaacgtaa cctatcccat

     4861 tacggtcaat ccgccgtttg ttcccacgga gaatccgacg ggttgttact cgctcacatt

     4921 taatgttgat gaaagctggc tacaggaagg ccagacgcga attatttttg atggcgttgg

     4981 aatt

//