請在線咨詢或直接聯系18986294316/17720555249

必騰預構載體定制最高100元/個

基本信息

質粒圖譜和多克隆位點信息

注意：

客戶在我公司選擇構建載體服務項目時，請自行提供母源載體。

如要使用本公司保存的母源載體時，本公司不能保證所有母源載體完全符合客戶預期，也不承擔由此產生的任何責任。

載體描述

pGEX-4T-1載體是一種常用的原核表達載體，具有Amp抗性，質粒大小為4.9 kb。該載體多克隆位點區域含有多個常用的內切酶位點序列，便于不同基因的克隆；且全序列中含有tac啟動子、GST標簽序列，是一種高效的蛋白表達載體。Tac啟動子在沒有誘導物存在的情況下，質粒上攜帶的lacIq基因產物可以有效抑制tac啟動子的轉錄。可在BL21、Rosstea等表達菌株中高表達融合有GST標簽的蛋白，且用凝血酶可將目的蛋白的GST標簽切掉，便于下游蛋白純化。

載體序列

LOCUS       Exported                4969 bp ds-DNA     circular SYN 14-MAR-2019

DEFINITION  synthetic circular DNA

ACCESSION   .

VERSION     .

KEYWORDS    .

SOURCE      synthetic DNA construct

  ORGANISM  synthetic DNA construct

REFERENCE   1  (bases 1 to 4969)

  AUTHORS   .

  TITLE     Direct Submission

  JOURNAL   Exported Thursday, Mar 14, 2019 from SnapGene Viewer 4.3.4

FEATURES             Location/Qualifiers

     source          1..4969

                     /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..935

                     /codon_start=1

                     /product="thrombin recognition and cleavage site"

                     /label=thrombin site

                     /translation="LVPRGS"

     misc_feature    936..966

                     /label=MCS

     primer_bind     complement(1019..1041)

                     /label=pGEX3'

     promoter        1272..1376

                     /gene="bla"

                     /label=AmpR promoter

     CDS             1377..2237

                     /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      2408..2996

                     /direction=RIGHT

                     /label=ori

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

                     replication"

     promoter        3240..3317

                     /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             3318..4400

                     /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"

     protein_bind    4413..4434

                     /label=CAP binding site

                     /bound_moiety="E. coli catabolite activator protein"

                     /note="CAP binding activates transcription in the presence

                     of cAMP."

     promoter        4449..4479

                     /label=lac promoter

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

     protein_bind    4487..4503

                     /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     4511..4527

                     /label=M13 rev

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

                     variants"

     CDS             4523..4969

                     /codon_start=1

                     /gene="lacZ fragment"

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

                     /label=lacZ-alpha

                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART

                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV

                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGI"

     primer_bind     complement(4543..4559)

                     /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 gttccgcgtg gatccccgga attcccgggt cgactcgagc

      961 ggccgcatcg tgactgactg acgatctgcc tcgcgcgttt cggtgatgac ggtgaaaacc

     1021 tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca

     1081 gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggcgca gccatgaccc

     1141 agtcacgtag cgatagcgga gtgtataatt cttgaagacg aaagggcctc gtgatacgcc

     1201 tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc

     1261 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc

     1321 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga

     1381 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt

     1441 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag

     1501 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag

     1561 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgtg

     1621 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg

     1681 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca

     1741 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag

     1801 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc

     1861 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg

     1921 cagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc

     1981 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg

     2041 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg

     2101 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga

     2161 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac

     2221 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa

     2281 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca

     2341 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag

     2401 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac

     2461 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa

     2521 ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc

     2581 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag

     2641 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac

     2701 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc

     2761 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc

     2821 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca

     2881 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc

     2941 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg

     3001 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct

     3061 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata

     3121 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc

     3181 gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc ataaattccg

     3241 acaccatcga atggtgcaaa acctttcgcg gtatggcatg atagcgcccg gaagagagtc

     3301 aattcagggt ggtgaatgtg aaaccagtaa cgttatacga tgtcgcagag tatgccggtg

     3361 tctcttatca gaccgtttcc cgcgtggtga accaggccag ccacgtttct gcgaaaacgc

     3421 gggaaaaagt ggaagcggcg atggcggagc tgaattacat tcccaaccgc gtggcacaac

     3481 aactggcggg caaacagtcg ttgctgattg gcgttgccac ctccagtctg gccctgcacg

     3541 cgccgtcgca aattgtcgcg gcgattaaat ctcgcgccga tcaactgggt gccagcgtgg

     3601 tggtgtcgat ggtagaacga agcggcgtcg aagcctgtaa agcggcggtg cacaatcttc

     3661 tcgcgcaacg cgtcagtggg ctgatcatta actatccgct ggatgaccag gatgccattg

     3721 ctgtggaagc tgcctgcact aatgttccgg cgttatttct tgatgtctct gaccagacac

     3781 ccatcaacag tattattttc tcccatgaag acggtacgcg actgggcgtg gagcatctgg

     3841 tcgcattggg tcaccagcaa atcgcgctgt tagcgggccc attaagttct gtctcggcgc

     3901 gtctgcgtct ggctggctgg cataaatatc tcactcgcaa tcaaattcag ccgatagcgg

     3961 aacgggaagg cgactggagt gccatgtccg gttttcaaca aaccatgcaa atgctgaatg

     4021 agggcatcgt tcccactgcg atgctggttg ccaacgatca gatggcgctg ggcgcaatgc

     4081 gcgccattac cgagtccggg ctgcgcgttg gtgcggatat ctcggtagtg ggatacgacg

     4141 ataccgaaga cagctcatgt tatatcccgc cgttaaccac catcaaacag gattttcgcc

     4201 tgctggggca aaccagcgtg gaccgcttgc tgcaactctc tcagggccag gcggtgaagg

     4261 gcaatcagct gttgcccgtc tcactggtga aaagaaaaac caccctggcg cccaatacgc

     4321 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc

     4381 gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca

     4441 ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa

     4501 caatttcaca caggaaacag ctatgaccat gattacggat tcactggccg tcgttttaca

     4561 acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc

     4621 tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg

     4681 cagcctgaat ggcgaatggc gctttgcctg gtttccggca ccagaagcgg tgccggaaag

     4741 ctggctggag tgcgatcttc ctgaggccga tactgtcgtc gtcccctcaa actggcagat

     4801 gcacggttac gatgcgccca tctacaccaa cgtaacctat cccattacgg tcaatccgcc

     4861 gtttgttccc acggagaatc cgacgggttg ttactcgctc acatttaatg ttgatgaaag

     4921 ctggctacag gaaggccaga cgcgaattat ttttgatggc gttggaatt

//