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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

pET32系列載體是設計用來克隆構建和高水平融合表達表達帶有109個氨基酸Trx標簽的蛋白質序列。將基因插入到載體的多克隆位點，能夠獲得融合表達蛋白質。同時融合蛋白包含有His標簽和S標簽，這些可以用于蛋白表達檢測和純化過程。

載體序列

LOCUS       Exported                5900 bp ds-DNA     circular SYN 14-MAR-2019

DEFINITION  synthetic circular DNA

ACCESSION   .

VERSION     .

KEYWORDS    pET32a

SOURCE      synthetic DNA construct

  ORGANISM  synthetic DNA construct

REFERENCE   1  (bases 1 to 5900)

  AUTHORS   .

  TITLE     Direct Submission

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

FEATURES             Location/Qualifiers

     source          1..5900

                     /organism="synthetic DNA construct"

                     /mol_type="other DNA"

     rep_origin      12..467

                     /direction=RIGHT

                     /label=f1 ori

                     /note="f1 bacteriophage origin of replication; arrow

                     indicates direction of (+) strand synthesis"

     promoter        494..598

                     /gene="bla"

                     /label=AmpR promoter

     CDS             599..1459

                     /codon_start=1

                     /gene="bla"

                     /product="beta-lactamase"

                     /label=AmpR

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

                     related antibiotics"

                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI

                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS

                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW

                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA

                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS

                     LIKHW"

     rep_origin      1630..2218

                     /direction=RIGHT

                     /label=ori

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

                     replication"

     misc_feature    2404..2546

                     /label=bom

                     /note="basis of mobility region from pBR322"

     CDS             complement(2648..2839)

                     /codon_start=1

                     /gene="rop"

                     /product="Rop protein, which maintains plasmids at low copy

                     number"

                     /label=rop

                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA

                     DELYRSCLARFGDDGENL"

     CDS             complement(3648..4730)

                     /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"

     promoter        complement(4731..4808)

                     /gene="lacI"

                     /label=lacI promoter

     primer_bind     5121..5140

                     /label=T7 primer

     promoter        5121..5139

                     /label=T7 promoter

                     /note="promoter for bacteriophage T7 RNA polymerase"

     protein_bind    5140..5164

                     /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)."

     RBS             5179..5201

                     /note="efficient ribosome binding site from bacteriophage

                     T7 gene 10 (Olins and Rangwala, 1989)"

     CDS             5209..5535

                     /codon_start=1

                     /gene="trxA"

                     /product="E. coli thioredoxin"

                     /label=TrxA

                     /translation="MSDKIIHLTDDSFDTDVLKADGAILVDFWAEWCGPCKMIAPILDE

                     IADEYQGKLTVAKLNIDQNPGTAPKYGIRGIPTLLLFKNGEVAATKVGALSKGQLKEFL

                     DANLA"

     CDS             5557..5574

                     /codon_start=1

                     /product="6xHis affinity tag"

                     /label=6xHis

                     /translation="HHHHHH"

     CDS             5584..5601

                     /codon_start=1

                     /product="thrombin recognition and cleavage site"

                     /label=thrombin site

                     /translation="LVPRGS"

     CDS             5608..5652

                     /codon_start=1

                     /product="affinity and epitope tag derived from pancreatic

                     ribonuclease A"

                     /label=S-Tag

                     /translation="KETAAAKFERQHMDS"

     CDS             5668..5682

                     /codon_start=1

                     /product="enterokinase recognition and cleavage site"

                     /label=enterokinase site

                     /translation="DDDDK"

     misc_feature    5683..5743

                     /label=MCS

     CDS             5744..5761

                     /codon_start=1

                     /product="6xHis affinity tag"

                     /label=6xHis

                     /translation="HHHHHH"

     primer_bind     complement(5814..5832)

                     /label=T7 ter primer

     terminator      5828..5875

                     /label=T7 terminator

                     /note="transcription terminator for bacteriophage T7 RNA

                     polymerase"

ORIGIN

        1 tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg

       61 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc

      121 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg

      181 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc

      241 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt

      301 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc

      361 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta

      421 acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaatttcag gtggcacttt

      481 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta

      541 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat

      601 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt

      661 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg

      721 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga

      781 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg

      841 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt

      901 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg

      961 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg

     1021 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga

     1081 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc

     1141 tgcagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc

     1201 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc

     1261 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg

     1321 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac

     1381 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc

     1441 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt

     1501 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac

     1561 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa

     1621 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc

     1681 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt

     1741 aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg

     1801 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc

     1861 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt

     1921 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga

     1981 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct

     2041 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg

     2101 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca

     2161 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa

     2221 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt

     2281 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga

     2341 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga

     2401 gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcatatatgg

     2461 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatac actccgctat

     2521 cgctacgtga ctgggtcatg gctgcgcccc gacacccgcc aacacccgct gacgcgccct

     2581 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct

     2641 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gaggcagctg cggtaaagct

     2701 catcagcgtg gtcgtgaagc gattcacaga tgtctgcctg ttcatccgcg tccagctcgt

     2761 tgagtttctc cagaagcgtt aatgtctggc ttctgataaa gcgggccatg ttaagggcgg

     2821 ttttttcctg tttggtcact gatgcctccg tgtaaggggg atttctgttc atgggggtaa

     2881 tgataccgat gaaacgagag aggatgctca cgatacgggt tactgatgat gaacatgccc

     2941 ggttactgga acgttgtgag ggtaaacaac tggcggtatg gatgcggcgg gaccagagaa

     3001 aaatcactca gggtcaatgc cagcgcttcg ttaatacaga tgtaggtgtt ccacagggta

     3061 gccagcagca tcctgcgatg cagatccgga acataatggt gcagggcgct gacttccgcg

     3121 tttccagact ttacgaaaca cggaaaccga agaccattca tgttgttgct caggtcgcag

     3181 acgttttgca gcagcagtcg cttcacgttc gctcgcgtat cggtgattca ttctgctaac

     3241 cagtaaggca accccgccag cctagccggg tcctcaacga caggagcacg atcatgcgca

     3301 cccgtggggc cgccatgccg gcgataatgg cctgcttctc gccgaaacgt ttggtggcgg

     3361 gaccagtgac gaaggcttga gcgagggcgt gcaagattcc gaataccgca agcgacaggc

     3421 cgatcatcgt cgcgctccag cgaaagcggt cctcgccgaa aatgacccag agcgctgccg

     3481 gcacctgtcc tacgagttgc atgataaaga agacagtcat aagtgcggcg acgatagtca

     3541 tgccccgcgc ccaccggaag gagctgactg ggttgaaggc tctcaagggc atcggtcgag

     3601 atcccggtgc ctaatgagtg agctaactta cattaattgc gttgcgctca ctgcccgctt

     3661 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag

     3721 gcggtttgcg tattgggcgc cagggtggtt tttcttttca ccagtgagac gggcaacagc

     3781 tgattgccct tcaccgcctg gccctgagag agttgcagca agcggtccac gctggtttgc

     3841 cccagcaggc gaaaatcctg tttgatggtg gttaacggcg ggatataaca tgagctgtct

     3901 tcggtatcgt cgtatcccac taccgagatg tccgcaccaa cgcgcagccc ggactcggta

     3961 atggcgcgca ttgcgcccag cgccatctga tcgttggcaa ccagcatcgc agtgggaacg

     4021 atgccctcat tcagcatttg catggtttgt tgaaaaccgg acatggcact ccagtcgcct

     4081 tcccgttccg ctatcggctg aatttgattg cgagtgagat atttatgcca gccagccaga

     4141 cgcagacgcg ccgagacaga acttaatggg cccgctaaca gcgcgatttg ctggtgaccc

     4201 aatgcgacca gatgctccac gcccagtcgc gtaccgtctt catgggagaa aataatactg

     4261 ttgatgggtg tctggtcaga gacatcaaga aataacgccg gaacattagt gcaggcagct

     4321 tccacagcaa tggcatcctg gtcatccagc ggatagttaa tgatcagccc actgacgcgt

     4381 tgcgcgagaa gattgtgcac cgccgcttta caggcttcga cgccgcttcg ttctaccatc

     4441 gacaccacca cgctggcacc cagttgatcg gcgcgagatt taatcgccgc gacaatttgc

     4501 gacggcgcgt gcagggccag actggaggtg gcaacgccaa tcagcaacga ctgtttgccc

     4561 gccagttgtt gtgccacgcg gttgggaatg taattcagct ccgccatcgc cgcttccact

     4621 ttttcccgcg ttttcgcaga aacgtggctg gcctggttca ccacgcggga aacggtctga

     4681 taagagacac cggcatactc tgcgacatcg tataacgtta ctggtttcac attcaccacc

     4741 ctgaattgac tctcttccgg gcgctatcat gccataccgc gaaaggtttt gcgccattcg

     4801 atggtgtccg ggatctcgac gctctccctt atgcgactcc tgcattagga agcagcccag

     4861 tagtaggttg aggccgttga gcaccgccgc cgcaaggaat ggtgcatgca aggagatggc

     4921 gcccaacagt cccccggcca cggggcctgc caccataccc acgccgaaac aagcgctcat

     4981 gagcccgaag tggcgagccc gatcttcccc atcggtgatg tcggcgatat aggcgccagc

     5041 aaccgcacct gtggcgccgg tgatgccggc cacgatgcgt ccggcgtaga ggatcgagat

     5101 cgatctcgat cccgcgaaat taatacgact cactataggg gaattgtgag cggataacaa

     5161 ttcccctcta gaaataattt tgtttaactt taagaaggag atatacatat gagcgataaa

     5221 attattcacc tgactgacga cagttttgac acggatgtac tcaaagcgga cggggcgatc

     5281 ctcgtcgatt tctgggcaga gtggtgcggt ccgtgcaaaa tgatcgcccc gattctggat

     5341 gaaatcgctg acgaatatca gggcaaactg accgttgcaa aactgaacat cgatcaaaac

     5401 cctggcactg cgccgaaata tggcatccgt ggtatcccga ctctgctgct gttcaaaaac

     5461 ggtgaagtgg cggcaaccaa agtgggtgca ctgtctaaag gtcagttgaa agagttcctc

     5521 gacgctaacc tggccggttc tggttctggc catatgcacc atcatcatca tcattcttct

     5581 ggtctggtgc cacgcggttc tggtatgaaa gaaaccgctg ctgctaaatt cgaacgccag

     5641 cacatggaca gcccagatct gggtaccgac gacgacgaca aggccatggc tgatatcgga

     5701 tccgaattcg agctccgtcg acaagcttgc ggccgcactc gagcaccacc accaccacca

     5761 ctgagatccg gctgctaaca aagcccgaaa ggaagctgag ttggctgctg ccaccgctga

     5821 gcaataacta gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgctgaa

     5881 aggaggaact atatccggat

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