| 出品公司: | Novagen (EMD Millipore |
|---|---|
| 载体名称: | pET-53-DEST |
| 质粒类型: | Gateway克隆载体 |
| 高拷贝/低拷贝: | 高拷贝 |
| 克隆方法: | Gateway |
| 启动子: | T7 |
| 载体大小: | 6832 bp |
| 5' 测序引物及序列: | T7: 5'-TAATACGACTCACTATAGGG-3' |
| 3' 测序引物及序列: | T7t: 5'-TGCTAGTTATTGCTCAGCGG-3' |
| 载体标签: | N-His |
| 载体抗性: | 氨苄青霉素 |
| 克隆菌株: | DH5a感受态细胞等 |
| 备注: | -- |
| 稳定性: | 否 |
| 组成型/诱导型: | 诱导型 |
| 病毒/非病毒: | 非病毒 |
| 表达宿主菌: | BL21(DE3) 、BLR(DE3)、Rosetta(DE3)、OrigamiB(DE3) |
| pET表达手册： |
pET表达手册英文版 pET表达手册中文版 |
| pET载体汇总: |
pET-3a pET-11a pET-12a pET-14b pET-15b pET-16b pET-17b pET-19b pET-20b pET-21a pET-22b pET-23a pET-24a pET-25b pET-26b pET-27b pET-28a pET-29a pET-30a pET-31b pET-32a pET-33b pET-34b pET-35b pET-37b pET-39b pET-40b pET-41a pET-42a pET-43.1a pET-44a pET-45b pET-47b pET-48b pET-49b pET-50b pET-51b pET-52b |
买家导航
pET-53-DEST载体 pET53-DEST质粒基本信息
pET-53-DEST质粒图谱 pET53-DEST载体图谱和pET-53-DEST载体序列 pET53-DEST质粒序列多克隆位点信息
pET-53-DEST质粒 pET53-DEST载体简介
pET-53-DEST质粒序列 pET53-DEST载体序列
LOCUS pET-53-DEST 6832 bp ds-DNA circular SYN 23-OCT-2018
DEFINITION Gateway(R) Nova bacterial destination vector for expressing proteins
tagged at the C-terminus with Strep-Tag II.
ACCESSION .
VERSION .
KEYWORDS pET-53-DEST
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6832)
AUTHORS MilliporeSigma (Novagen)
TITLE Direct Submission
JOURNAL Exported Monday, February 10, 2020 from SnapGene Viewer 3.1.4
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6832
/organism="synthetic DNA construct"
/lab_host="Escherichia coli"
/mol_type="other DNA"
rep_origin 12..467
/direction=RIGHT
/note="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
CDS complement(582..1442)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/note="AmpR"
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(1443..1535)
/gene="bla"
/note="AmpR promoter"
rep_origin 1616..2204
/direction=RIGHT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(2634..2825)
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/note="rop"
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
CDS complement(3397..4479)
/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"
promoter complement(4480..4557)
/gene="lacI"
/note="lacI promoter"
promoter 4870..4888
/note="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 4889..4913
/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)."
RBS 4944..4949
/note="ribosome binding site"
CDS 4957..4959
/codon_start=1
/product="start codon"
/note="ATG"
/translation="M"
CDS 4963..4980
/codon_start=1
/product="6xHis affinity tag"
/note="6xHis"
/translation="HHHHHH"
protein_bind 4984..5108
/gene="mutant version of attR"
/bound_moiety="LR Clonase(TM)"
/note="attR1"
/note="recombination site for the Gateway(R) LR reaction"
promoter 5133..5163
/note="lac UV5 promoter"
/note="E. coli lac promoter with an ""up"" mutation"
CDS 5217..5876
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/note="CmR"
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
CDS 6218..6523
/codon_start=1
/gene="ccdB"
/product="CcdB, a bacterial toxin that poisons DNA gyrase"
/note="ccdB"
/note="Plasmids containing the ccdB gene cannot be
propagated in standard E. coli strains."
/translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
protein_bind complement(6564..6688)
/gene="mutant version of attR"
/bound_moiety="LR Clonase(TM)"
/note="attR2"
/note="recombination site for the Gateway(R) LR reaction"
CDS 6696..6719
/codon_start=1
/product="peptide that binds Strep-Tactin, an engineered
form of streptavidin"
/note="Strep-Tag II"
/translation="WSHPQFEK"
terminator 6760..6807
/note="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 acaatttctg gcggcacgat
481 ggcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
541 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
601 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
661 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
721 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
781 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
841 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
901 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
961 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
1021 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
1081 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
1141 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
1201 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
1261 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
1321 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
1381 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
1441 atactcttcc tttttcaatc atgattgaag catttatcag ggttattgtc tcatgagcgg
1501 atacatattt gaatgtattt agaaaaataa acaaataggt catgaccaaa atcccttaac
1561 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
1621 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
1681 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
1741 gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga
1801 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
1861 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
1921 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
1981 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
2041 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
2101 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
2161 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
2221 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
2281 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
2341 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt
2401 attttctcct tacgcatctg tgcggtattt cacaccgcat atatggtgca ctctcagtac
2461 aatctgctct gatgccgcat agttaagcca gtatacactc cgctatcgct acgtgactgg
2521 gtcatggctg cgccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg
2581 ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg
2641 ttttcaccgt catcaccgaa acgcgcgagg cagctgcggt aaagctcatc agcgtggtcg
2701 tgaagcgatt cacagatgtc tgcctgttca tccgcgtcca gctcgttgag tttctccaga
2761 agcgttaatg tctggcttct gataaagcgg gccatgttaa gggcggtttt ttcctgtttg
2821 gtcactgatg cctccgtgta agggggattt ctgttcatgg gggtaatgat accgatgaaa
2881 cgagagagga tgctcacgat acgggttact gatgatgaac atgcccggtt actggaacgt
2941 tgtgagggta aacaactggc ggtatggatg cggcgggacc agagaaaaat cactcagggt
3001 caatgccagc gcttcgttaa tacagatgta ggtgttccac agggtagcca gcagcatcct
3061 gcgatgcaga tccggaacat aatggtgcag ggcgctgact tccgcgtttc cagactttac
3121 gaaacacgga aaccgaagac cattcatgtt gttgctcagg tcgcagacgt tttgcagcag
3181 cagtcgcttc acgttcgctc gcgtatcggt gattcattct gctaaccagt aaggcaaccc
3241 cgccagccta gccgggtcct caacgacagg agcacgatca tgctagtcat gccccgcgcc
3301 caccggaagg agctgactgg gttgaaggct ctcaagggca tcggtcgaga tcccggtgcc
3361 taatgagtga gctaacttac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
3421 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
3481 attgggcgcc agggtggttt ttcttttcac cagtgagacg ggcaacagct gattgccctt
3541 caccgcctgg ccctgagaga gttgcagcaa gcggtccacg ctggtttgcc ccagcaggcg
3601 aaaatcctgt ttgatggtgg ttaacggcgg gatataacat gagctgtctt cggtatcgtc
3661 gtatcccact accgagatgt ccgcaccaac gcgcagcccg gactcggtaa tggcgcgcat
3721 tgcgcccagc gccatctgat cgttggcaac cagcatcgca gtgggaacga tgccctcatt
3781 cagcatttgc atggtttgtt gaaaaccgga catggcactc cagtcgcctt cccgttccgc
3841 tatcggctga atttgattgc gagtgagata tttatgccag ccagccagac gcagacgcgc
3901 cgagacagaa cttaatgggc ccgctaacag cgcgatttgc tggtgaccca atgcgaccag
3961 atgctccacg cccagtcgcg taccgtcttc atgggagaaa ataatactgt tgatgggtgt
4021 ctggtcagag acatcaagaa ataacgccgg aacattagtg caggcagctt ccacagcaat
4081 ggcatcctgg tcatccagcg gatagttaat gatcagccca ctgacgcgtt gcgcgagaag
4141 attgtgcacc gccgctttac aggcttcgac gccgcttcgt tctaccatcg acaccaccac
4201 gctggcaccc agttgatcgg cgcgagattt aatcgccgcg acaatttgcg acggcgcgtg
4261 cagggccaga ctggaggtgg caacgccaat cagcaacgac tgtttgcccg ccagttgttg
4321 tgccacgcgg ttgggaatgt aattcagctc cgccatcgcc gcttccactt tttcccgcgt
4381 tttcgcagaa acgtggctgg cctggttcac cacgcgggaa acggtctgat aagagacacc
4441 ggcatactct gcgacatcgt ataacgttac tggtttcaca ttcaccaccc tgaattgact
4501 ctcttccggg cgctatcatg ccataccgcg aaaggttttg cgccattcga tggtgtccgg
4561 gatctcgacg ctctccctta tgcgactcct gcattaggaa gcagcccagt agtaggttga
4621 ggccgttgag caccgccgcc gcaaggaatg gtgcatgcaa ggagatggcg cccaacagtc
4681 ccccggccac ggggcctgcc accataccca cgccgaaaca agcgctcatg agcccgaagt
4741 ggcgagcccg atcttcccca tcggtgatgt cggcgatata ggcgccagca accgcacctg
4801 tggcgccggt gatgccggcc acgatgcgtc cggcgtagag gatcgagatc gatctcgatc
4861 ccgcgaaatt aatacgactc actatagggg aattgtgagc ggataacaat tcccctctag
4921 aaataatttt gtttaacttt aagaaggaga tataccatgg cacatcacca ccaccatcac
4981 gtcacaagtt tgtacaaaaa agctgaacga gaaacgtaaa atgatataaa tatcaatata
5041 ttaaattaga ttttgcataa aaaacagact acataatact gtaaaacaca acatatccag
5101 tcactatggc ggccgcatta ggcaccccag gctttacact ttatgcttcc ggctcgtata
5161 atgtgtggat tttgagttag gatccgtcga gattttcagg agctaaggaa gctaaaatgg
5221 agaaaaaaat cactggatat accaccgttg atatatccca atggcatcgt aaagaacatt
5281 ttgaggcatt tcagtcagtt gctcaatgta cctataacca gaccgttcag ctggatatta
5341 cggccttttt aaagaccgta aagaaaaata agcacaagtt ttatccggcc tttattcaca
5401 ttcttgcccg cctgatgaat gctcatccgg aattccgtat ggcaatgaaa gacggtgagc
5461 tggtgatatg ggatagtgtt cacccttgtt acaccgtttt ccatgagcaa actgaaacgt
5521 tttcatcgct ctggagtgaa taccacgacg atttccggca gtttctacac atatattcgc
5581 aagatgtggc gtgttacggt gaaaacctgg cctatttccc taaagggttt attgagaata
5641 tgtttttcgt ctcagccaat ccctgggtga gtttcaccag ttttgattta aacgtggcca
5701 atatggacaa cttcttcgcc cccgttttca ctatgggcaa atattatacg caaggcgaca
5761 aggtgctgat gccgctggcg attcaggttc atcatgccgt ttgtgatggc ttccatgtcg
5821 gcagaatgct taatgaatta caacagtact gcgatgagtg gcagggcggg gcgtaaacgc
5881 gtggatccgg cttactaaaa gccagataac agtatgcgta tttgcgcgct gatttttgcg
5941 gtataagaat atatactgat atgtataccc gaagtatgtc aaaaagaggt atgctatgaa
6001 gcagcgtatt acagtgacag ttgacagcga cagctatcag ttgctcaagg catatatgat
6061 gtcaatatct ccggtctggt aagcacaacc atgcagaatg aagcccgtcg tctgcgtgcc
6121 gaacgctgga aagcggaaaa tcaggaaggg atggctgagg tcgcccggtt tattgaaatg
6181 aacggctctt ttgctgacga gaacaggggc tggtgaaatg cagtttaagg tttacaccta
6241 taaaagagag agccgttatc gtctgtttgt ggatgtacag agtgatatta ttgacacgcc
6301 cgggcgacgg atggtgatcc ccctggccag tgcacgtctg ctgtcagata aagtctcccg
6361 tgaactttac ccggtggtgc atatcgggga tgaaagctgg cgcatgatga ccaccgatat
6421 ggccagtgtg ccggtttccg ttatcgggga agaagtggct gatctcagcc accgcgaaaa
6481 tgacatcaaa aacgccatta acctgatgtt ctggggaata taaatgtcag gctcccttat
6541 acacagccag tctgcaggtc gaccatagtg actggatatg ttgtgtttta cagtattatg
6601 tagtctgttt tttatgcaaa atctaattta atatattgat atttatatca ttttacgttt
6661 ctcgttcagc tttcttgtac aaagtggttt ccgcttggag ccacccgcag ttcgaaaagt
6721 aagtgattaa cctaggctgc tgccaccgct gagcaataac tagcataacc ccttggggcc
6781 tctaaacggg tcttgagggg ttttttgctg aaaggaggaa ctatatccgg at
//
