pET-28a质粒试用

产品信息：
名称：pET-28a
品牌：Novagen
货号：Vector10129
规格：1ug质粒(10ul,100ng/ul),500ul甘油菌
储存温度：-20℃
产品简介：
pET-28a载体是大肠杆菌表达载体,带有Kanamycin (卡那霉素)抗性，启动子是T7，复制子是Ori,质粒大小是5369bp，标签是N-His, N-Thrombin, N-T7, C-His可转化进大肠杆菌中大量复制 (37℃,LB)。
5' 测序引物:T7：TAATACGACTCACTATAGGG
3' 测序引物:T7-Ter：TGCTAGTTATTGCTCAGCGG
载体图谱：



操作说明：
本产品包含一份质粒和一份含该质粒的克隆菌种，收到产品后请取1ul质粒转化，不建议优先使用菌种，如需使用菌种，请四区划线后挑单菌落培养。
载体序列：
LOCUS Exported 5369 bp ds-DNA circular SYN 31-7-2015
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS Untitled
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5369)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported 2015-7-31 MLCC
http://www.miaolingbio.com
FEATURES Location/Qualifiers
source 1..5369
/organism="synthetic DNA construct"
/mol_type="other DNA"
source 306..328
/organism="Enterobacteria phage T7"
/mol_type="genomic DNA"
/db_xref="taxon:10760"
terminator 26..73
/note="T7 terminator"
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
CDS complement(140..157)
/codon_start=1
/product="6xHis affinity tag"
/note="6xHis"
/translation="HHHHHH"
CDS complement(207..239)
/codon_start=1
/product="leader peptide from bacteriophage T7 gene 10"
/note="T7 tag (gene 10 leader)"
/note="promotes efficient translation in E. coli"
/translation="MASMTGGQQMG"
CDS complement(243..260)
/codon_start=1
/product="thrombin recognition and cleavage site"
/note="thrombin site"
/translation="LVPRGS"
CDS complement(270..287)
/codon_start=1
/product="6xHis affinity tag"
/note="6xHis"
/translation="HHHHHH"
RBS 306..328
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
protein_bind 343..367
/bound_moiety="lac repressor encoded by lacI"
/note="lac operator"
/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(368..386)
/note="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
promoter 695..772
/gene="lacI"
/note="lacI promoter"
CDS 773..1855
/codon_start=1
/gene="lacI"
/product="lac repressor"
/note="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"
CDS 2664..2855
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/note="rop"
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
misc_feature 2957..3099
/note="bom"
/note="basis of mobility region from pBR322"
rep_origin complement(3285..3873)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS 3995..4810
/codon_start=1
/product="aminoglycoside phosphotransferase"
/note="KanR"
/note="confers resistance to kanamycin in bacteria or G418
(Geneticin(R)) in eukaryotes"
/translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
rep_origin complement(4903..5358)
/direction=LEFT
/note="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 gagtgcggcc gcaagcttgt
181 cgacggagct cgaattcgga tccgcgaccc atttgctgtc caccagtcat gctagccata
241 tggctgccgc gcggcaccag gccgctgctg tgatgatgat gatgatggct gctgcccatg
301 gtatatctcc ttcttaaagt taaacaaaat tatttctaga ggggaattgt tatccgctca
361 caattcccct atagtgagtc gtattaattt cgcgggatcg agatctcgat cctctacgcc
421 ggacgcatcg tggccggcat caccggcgcc acaggtgcgg ttgctggcgc ctatatcgcc
481 gacatcaccg atggggaaga tcgggctcgc cacttcgggc tcatgagcgc ttgtttcggc
541 gtgggtatgg tggcaggccc cgtggccggg ggactgttgg gcgccatctc cttgcatgca
601 ccattccttg cggcggcggt gctcaacggc ctcaacctac tactgggctg cttcctaatg
661 caggagtcgc ataagggaga gcgtcgagat cccggacacc atcgaatggc gcaaaacctt
721 tcgcggtatg gcatgatagc gcccggaaga gagtcaattc agggtggtga atgtgaaacc
781 agtaacgtta tacgatgtcg cagagtatgc cggtgtctct tatcagaccg tttcccgcgt
841 ggtgaaccag gccagccacg tttctgcgaa aacgcgggaa aaagtggaag cggcgatggc
901 ggagctgaat tacattccca accgcgtggc acaacaactg gcgggcaaac agtcgttgct
961 gattggcgtt gccacctcca gtctggccct gcacgcgccg tcgcaaattg tcgcggcgat
1021 taaatctcgc gccgatcaac tgggtgccag cgtggtggtg tcgatggtag aacgaagcgg
1081 cgtcgaagcc tgtaaagcgg cggtgcacaa tcttctcgcg caacgcgtca gtgggctgat
1141 cattaactat ccgctggatg accaggatgc cattgctgtg gaagctgcct gcactaatgt
1201 tccggcgtta tttcttgatg tctctgacca gacacccatc aacagtatta ttttctccca
1261 tgaagacggt acgcgactgg gcgtggagca tctggtcgca ttgggtcacc agcaaatcgc
1321 gctgttagcg ggcccattaa gttctgtctc ggcgcgtctg cgtctggctg gctggcataa
1381 atatctcact cgcaatcaaa ttcagccgat agcggaacgg gaaggcgact ggagtgccat
1441 gtccggtttt caacaaacca tgcaaatgct gaatgagggc atcgttccca ctgcgatgct
1501 ggttgccaac gatcagatgg cgctgggcgc aatgcgcgcc attaccgagt ccgggctgcg
1561 cgttggtgcg gatatctcgg tagtgggata cgacgatacc gaagacagct catgttatat
1621 cccgccgtta accaccatca aacaggattt tcgcctgctg gggcaaacca gcgtggaccg
1681 cttgctgcaa ctctctcagg gccaggcggt gaagggcaat cagctgttgc ccgtctcact
1741 ggtgaaaaga aaaaccaccc tggcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
1801 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
1861 acgcaattaa tgtaagttag ctcactcatt aggcaccggg atctcgaccg atgcccttga
1921 gagccttcaa cccagtcagc tccttccggt gggcgcgggg catgactatc gtcgccgcac
1981 ttatgactgt cttctttatc atgcaactcg taggacaggt gccggcagcg ctctgggtca
2041 ttttcggcga ggaccgcttt cgctggagcg cgacgatgat cggcctgtcg cttgcggtat
2101 tcggaatctt gcacgccctc gctcaagcct tcgtcactgg tcccgccacc aaacgtttcg
2161 gcgagaagca ggccattatc gccggcatgg cggccccacg ggtgcgcatg atcgtgctcc
2221 tgtcgttgag gacccggcta ggctggcggg gttgccttac tggttagcag aatgaatcac
2281 cgatacgcga gcgaacgtga agcgactgct gctgcaaaac gtctgcgacc tgagcaacaa
2341 catgaatggt cttcggtttc cgtgtttcgt aaagtctgga aacgcggaag tcagcgccct
2401 gcaccattat gttccggatc tgcatcgcag gatgctgctg gctaccctgt ggaacaccta
2461 catctgtatt aacgaagcgc tggcattgac cctgagtgat ttttctctgg tcccgccgca
2521 tccataccgc cagttgttta ccctcacaac gttccagtaa ccgggcatgt tcatcatcag
2581 taacccgtat cgtgagcatc ctctctcgtt tcatcggtat cattaccccc atgaacagaa
2641 atccccctta cacggaggca tcagtgacca aacaggaaaa aaccgccctt aacatggccc
2701 gctttatcag aagccagaca ttaacgcttc tggagaaact caacgagctg gacgcggatg
2761 aacaggcaga catctgtgaa tcgcttcacg accacgctga tgagctttac cgcagctgcc
2821 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
2881 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
2941 ttggcgggtg tcggggcgca gccatgaccc agtcacgtag cgatagcgga gtgtatactg
3001 gcttaactat gcggcatcag agcagattgt actgagagtg caccatatat gcggtgtgaa
3061 ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gctcttccgc ttcctcgctc
3121 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
3181 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
3241 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
3301 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
3361 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
3421 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
3481 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
3541 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
3601 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
3661 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
3721 agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
3781 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
3841 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
3901 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgaa caataaaact
3961 gtctgcttac ataaacagta atacaagggg tgttatgagc catattcaac gggaaacgtc
4021 ttgctctagg ccgcgattaa attccaacat ggatgctgat ttatatgggt ataaatgggc
4081 tcgcgataat gtcgggcaat caggtgcgac aatctatcga ttgtatggga agcccgatgc
4141 gccagagttg tttctgaaac atggcaaagg tagcgttgcc aatgatgtta cagatgagat
4201 ggtcagacta aactggctga cggaatttat gcctcttccg accatcaagc attttatccg
4261 tactcctgat gatgcatggt tactcaccac tgcgatcccc gggaaaacag cattccaggt
4321 attagaagaa tatcctgatt caggtgaaaa tattgttgat gcgctggcag tgttcctgcg
4381 ccggttgcat tcgattcctg tttgtaattg tccttttaac agcgatcgcg tatttcgtct
4441 cgctcaggcg caatcacgaa tgaataacgg tttggttgat gcgagtgatt ttgatgacga
4501 gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg cataaacttt tgccattctc
4561 accggattca gtcgtcactc atggtgattt ctcacttgat aaccttattt ttgacgaggg
4621 gaaattaata ggttgtattg atgttggacg agtcggaatc gcagaccgat accaggatct
4681 tgccatccta tggaactgcc tcggtgagtt ttctccttca ttacagaaac ggctttttca
4741 aaaatatggt attgataatc ctgatatgaa taaattgcag tttcatttga tgctcgatga
4801 gtttttctaa gaattaattc atgagcggat acatatttga atgtatttag aaaaataaac
4861 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgaaattgta aacgttaata
4921 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg
4981 aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc
5041 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa
5101 ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt
5161 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac
5221 ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta
5281 gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg
5341 cgccgctaca gggcgcgtcc cattcgcca
//
转自：http://www.miaolingbio.com/cn/product/plasmid/434.html