• 规格：20μl质粒
• 启动子：T7/lac promoter
• 复制子：ori，F1 ori
• 终止子：T7 terminator
• 质粒分类：基因文库质粒；其它基因质粒；其它种源质粒
• 质粒大小：7297bp
• 质粒标签：C-6×His
• 原核抗性：卡那霉素Kan（50μg/ml）
• 克隆菌株：DH5α等大肠杆菌
• 表达宿主：BL21(DE3)大肠杆菌
• 诱导方式：IPTG或乳糖及其类似物
• 质粒简介：             pET28a-PPK质粒可以转化到含DE3的大肠杆菌中，通过IPTG的诱导表达PPK基因。
• 质粒图谱1： png/1-1FQ5153622148.png

质粒序列： 点击查看序列

LOCUS       Exported                7297 bp ds-DNA     circular SYN 15-AUG-2017

DEFINITION  synthetic circular DNA

FEATURES             Location/Qualifiers

     source          1..7297

                     /organism="synthetic DNA construct"

                     /mol_type="other DNA"

     terminator      26..73

                     /label=T7 terminator

                     /note="transcription terminator for bacteriophage T7 RNA 

                     polymerase"

     CDS             complement(140..157)

                     /codon_start=1

                     /product="6xHis affinity tag"

                     /label=6xHis

                     /translation="HHHHHH"

     misc_feature    complement(164..2227)

                     /label=PPK

     RBS             2234..2256

                     /note="efficient ribosome binding site from bacteriophage 

                     T7 gene 10 (Olins and Rangwala, 1989)"

     protein_bind    2271..2295

                     /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)."

     promoter        complement(2296..2314)

                     /label=T7 promoter

                     /note="promoter for bacteriophage T7 RNA polymerase"

     promoter        2623..2700

                     /gene="lacI"

                     /label=lacI promoter

     CDS             2701..3783

                     /codon_start=1

                     /gene="lacI"

                     /product="lac repressor"

                     /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"

     CDS             4592..4783

                     /codon_start=1

                     /gene="rop"

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

                     number"

                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA

                     DELYRSCLARFGDDGENL"

     misc_feature    4885..5027

                     /label=bom

                     /note="basis of mobility region from pBR322"

     rep_origin      complement(5213..5801)

                     /direction=LEFT

                     /label=ori

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

                     replication"

     CDS             5923..6738

                     /codon_start=1

                     /gene="aph(3')-Ia"

                     /product="aminoglycoside phosphotransferase"

                     /label=KanR

                     /note="confers resistance to kanamycin in bacteria or G418 

                     (Geneticin(R)) in eukaryotes"

                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG

                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK

                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA

                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI

                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"

     rep_origin      complement(6831..7286)

                     /direction=LEFT

                     /label=f1 ori

                     /note="f1 bacteriophage origin of replication; arrow 

                     indicates direction of (+) strand synthesis"

ORIGIN

        1 atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa

       61 ggggttatgc tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt

      121 tgttagcagc cggatctcag tggtggtggt ggtggtgctc gagttcaggt tgttcaagag

      181 atttgatgta atcgtagata gcaagttgag cacgaacttt acgacggttg ccacgaggaa

      241 cgtaacggtt agaaagttct ttatcgatgt aacgagcttt aactgtatca gagaaaagga

      301 tatcgatgat atcaagaaca cgttgtttaa gacgaggatc aagaagaggt gtagcaactt

      361 cgatacggta atcgatgtta cgtgtcatcc aatcagcaga agaaaggtaa acttttttat

      421 cgccgccgtt ttcgaagatg taaacacgat catgttcaag gtaacgatca acgatagaga

      481 tagcacggat gttatcagag atgccttcaa ggttagggat aagagagcac atgccacgaa

      541 caagaaggtt aacaggaacg ccagaagaag aagcagcgta aagacgatca acaaggcctt

      601 tatcaacaag gttgttaagt ttaagtgtga tgccagaagg aaggccttgt tgagcgttag

      661 cgatttcacg atcaaccatt tcgtaaagaa gacgacgaga gttttgagga gaaaccataa

      721 ggtaatcgaa tgtaacagga cggtaagggt tttcgatgaa gttgaaaaca cgacgaactt

      781 cgtttgtgat acgagcatca gctgtaagaa gagagtaatc tgtgtaaaga cgagctgttt

      841 tttcgttgaa gttgcctgtg ccgatatgag cgtaacgaac aacttcgccg ttttctttac

      901 gagagataag gaaaagttta gcatggattt taaggccagg agcagagaag ataacatgaa

      961 cgccagcttc tgtaagacgt ttagcccaat ggatgttagc ttcttcatcg aaacgagctt

     1021 gaagttcaac aacaactgta acttttttgc cgttatgagc agcatggatc atagaatcga

     1081 tgatacgaga atctttagca acacggtaga tgttgatttt gatagcaaga acagaaggat

     1141 cgaaagaagc ttgacgaaga agttcaagaa catgttcgaa tgtatggtaa gggtagtaaa

     1201 gaagaacatc acgttcacgg atagcatcga agccgttacg gaattgagct ttatcgaacc

     1261 agatatgacg aagacgagga agaggtttgt taacaaggtt agctttgcca acgttaggga

     1321 agttgatgaa atctttgaag ttatggtaac ggccgccagg aacgatagaa tcgtaacgag

     1381 agattgtaag tttttcacga agaacttcaa caagagcgtt aggcatatca cgttggtaaa

     1441 cgaaacgaac aggttcagct gtaagacgtt gtttaagaga agaagacata agttccataa

     1501 gagaagcttc catttcatga acaagatcgt attcagcatc acgtgtcatt ttcatagagt

     1561 aagcgttaag agcatcgtaa tcgaagaagc ctttgaagat atcatcaagg cagtaacgaa

     1621 ggatgttatc aagaaggatc ataggtttac gacgacgagg agcttcagga ggaaggttaa

     1681 cgaaacgagg aactttatca gaagggattt caagaagagc gtaacggatt gtatcgccac

     1741 ggatgatttc aacagcaagg tatgtgtaat catctttaag gaattgaaca agatctgtat

     1801 cagggttgat aaggataggt gtgatatgtt gacgaaggta ttgtttgaag taatgacgaa

     1861 gccagttttg ttggttaaca gaaagttgac gttcgttgat aaggaagatt tggttacgag

     1921 ccatttcaag aagaagttcg ttgtaaaggc catcgaattc ttgatcagct ttaagaacac

     1981 gagattggat tttgccaaga agatgacgag aatgagagtt agagccttgt tcttcagaga

     2041 tgatgatacg acgtttaagt tcagcgaaac gaactttgta gaattcatca aggttgttag

     2101 agtagatgcc aaggaaacgc atacgttcga taagagggtt agatttatca gcagcttctt

     2161 gaagaacacg ttcgttgaaa gaaagccaag aaagttcttt ttcgatgtaa agtttttctt

     2221 ggcccatggt atatctcctt cttaaagtta aacaaaatta tttctagagg ggaattgtta

     2281 tccgctcaca attcccctat agtgagtcgt attaatttcg cgggatcgag atctcgatcc

     2341 tctacgccgg acgcatcgtg gccggcatca ccggcgccac aggtgcggtt gctggcgcct

     2401 atatcgccga catcaccgat ggggaagatc gggctcgcca cttcgggctc atgagcgctt

     2461 gtttcggcgt gggtatggtg gcaggccccg tggccggggg actgttgggc gccatctcct

     2521 tgcatgcacc attccttgcg gcggcggtgc tcaacggcct caacctacta ctgggctgct

     2581 tcctaatgca ggagtcgcat aagggagagc gtcgagatcc cggacaccat cgaatggcgc

     2641 aaaacctttc gcggtatggc atgatagcgc ccggaagaga gtcaattcag ggtggtgaat

     2701 gtgaaaccag taacgttata cgatgtcgca gagtatgccg gtgtctctta tcagaccgtt

     2761 tcccgcgtgg tgaaccaggc cagccacgtt tctgcgaaaa cgcgggaaaa agtggaagcg

     2821 gcgatggcgg agctgaatta cattcccaac cgcgtggcac aacaactggc gggcaaacag

     2881 tcgttgctga ttggcgttgc cacctccagt ctggccctgc acgcgccgtc gcaaattgtc

     2941 gcggcgatta aatctcgcgc cgatcaactg ggtgccagcg tggtggtgtc gatggtagaa

     3001 cgaagcggcg tcgaagcctg taaagcggcg gtgcacaatc ttctcgcgca acgcgtcagt

     3061 gggctgatca ttaactatcc gctggatgac caggatgcca ttgctgtgga agctgcctgc

     3121 actaatgttc cggcgttatt tcttgatgtc tctgaccaga cacccatcaa cagtattatt

     3181 ttctcccatg aagacggtac gcgactgggc gtggagcatc tggtcgcatt gggtcaccag

     3241 caaatcgcgc tgttagcggg cccattaagt tctgtctcgg cgcgtctgcg tctggctggc

     3301 tggcataaat atctcactcg caatcaaatt cagccgatag cggaacggga aggcgactgg

     3361 agtgccatgt ccggttttca acaaaccatg caaatgctga atgagggcat cgttcccact

     3421 gcgatgctgg ttgccaacga tcagatggcg ctgggcgcaa tgcgcgccat taccgagtcc

     3481 gggctgcgcg ttggtgcgga tatctcggta gtgggatacg acgataccga agacagctca

     3541 tgttatatcc cgccgttaac caccatcaaa caggattttc gcctgctggg gcaaaccagc

     3601 gtggaccgct tgctgcaact ctctcagggc caggcggtga agggcaatca gctgttgccc

     3661 gtctcactgg tgaaaagaaa aaccaccctg gcgcccaata cgcaaaccgc ctctccccgc

     3721 gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag

     3781 tgagcgcaac gcaattaatg taagttagct cactcattag gcaccgggat ctcgaccgat

     3841 gcccttgaga gccttcaacc cagtcagctc cttccggtgg gcgcggggca tgactatcgt

     3901 cgccgcactt atgactgtct tctttatcat gcaactcgta ggacaggtgc cggcagcgct

     3961 ctgggtcatt ttcggcgagg accgctttcg ctggagcgcg acgatgatcg gcctgtcgct

     4021 tgcggtattc ggaatcttgc acgccctcgc tcaagccttc gtcactggtc ccgccaccaa

     4081 acgtttcggc gagaagcagg ccattatcgc cggcatggcg gccccacggg tgcgcatgat

     4141 cgtgctcctg tcgttgagga cccggctagg ctggcggggt tgccttactg gttagcagaa

     4201 tgaatcaccg atacgcgagc gaacgtgaag cgactgctgc tgcaaaacgt ctgcgacctg

     4261 agcaacaaca tgaatggtct tcggtttccg tgtttcgtaa agtctggaaa cgcggaagtc