| 质粒名称: | pET His6GST TEV LIC cloning vector |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 29655 |
| 存储实验室: | Vladislav Verkhusha |
|
载体骨架基本信息 |
|
| 载体名称: | pET |
| 载体出品公司: | Clontech |
| 空载体大小: | 6015 bp |
| 完整质粒大小: | 6015 bp |
| 载体类型: | 细菌表达 |
| 载体抗性: | Kanamycin, 50 μg/mL |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 低拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | 无 |
| 标签/融合蛋白: |
6xHis (N terminal on backbone)
GST (N terminal on backbone)
TEV cleavage site (N terminal on backbone)
|
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | LIC tag (destroyed during cloning) |
| 3’克隆位点: | LIC tag (destroyed during cloning) |
| 5’测序引物 | GST forward |
| 3’测序引物 | T7 reverse |
|
文献引用方法 |
|
| 材料和方法部分: | pET His6 GST TEV LIC cloning vector (1G) was a gift from Scott Gradia (Addgene plasmid # 29655 ; http://n2t.net/addgene:29655 ; RRID:Addgene_29655) |
买家导航
pET His6GST TEV LIC cloning vector载体质粒基本信息
pET His6GST TEV LIC cloning vector质粒图谱载体图谱和 pET His6GST TEV LIC cloning vector载体序列质粒序列多克隆位点信息
pET His6GST TEV LIC cloning vector质粒载体简介
pET His6GST TEV LIC cloning vector质粒序列载体序列
LOCUS pET His6 GST TEV LIC cloning vector 6012 bp DNA circular SYN 14-APR-2025
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6012)
TITLE LIC N-terminal fusion vectors for E. coli expression
REFERENCE 2 (bases 1 to 6012)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Apr 14, 2025 from SnapGene Server 8.0.1
https://www.biofeng.com
COMMENT SGRef: number: 1; type: "Journal Article"
COMMENT Sequence Label: pET His6 GST TEV LIC cloning vector (1G)
FEATURES Location/Qualifiers
source 1..6012
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind 67..86
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
primer_bind 170..189
/label=T7
/note="T7 promoter, forward primer"
promoter 170..188
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 189..213
/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 228..250
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 269..286
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
CDS 296..949
/codon_start=1
/product="glutathione S-transferase from Schistosoma
japonicum"
/label=GST
/translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
CDS 959..979
/codon_start=1
/product="tobacco etch virus (TEV) protease recognition and
cleavage site"
/label=TEV site
/translation="ENLYFQS"
CDS 1045..1062
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
primer_bind complement(1115..1133)
/label=T7 Term
/note="T7 terminator, reverse primer"
terminator 1129..1176
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
rep_origin 1213..1668
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(1300..1319)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 1510..1531
/label=F1ori-F
/note="F1 origin, forward primer"
CDS complement(1761..2576)
/codon_start=1
/product="aminoglycoside phosphotransferase"
/label=KanR
/note="confers resistance to kanamycin in bacteria or G418
(Geneticin(R)) in eukaryotes"
/translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
CDS complement(1761..2576)
/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"
primer_bind 2484..2503
/label=Kan-R
/note="Kanamycin resistance gene, reverse primer"
primer_bind 2568..2587
/label=pENTR-R
/note="pENTR vectors, reverse primer"
rep_origin 2698..3286
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 3187..3206
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 3440..3457
/label=L4440
/note="L4440 vector, forward primer"
misc_feature 3472..3611
/label=bom
/note="basis of mobility region from pBR322"
primer_bind complement(3537..3556)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 3697..3719
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
CDS complement(3713..3904)
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/label=rop
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
CDS complement(4713..5795)
/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"
primer_bind 5757..5776
/label=LacI-R
/note="LacI, reverse primer"
promoter complement(5796..5873)
/gene="lacI"
/label=lacI promoter
ORIGIN
1 tgccaccata cccacgccga aacaagcgct catgagcccg aagtggcgag cccgatcttc
61 cccatcggtg atgtcggcga tataggcgcc agcaaccgca cctgtggcgc cggtgatgcc
121 ggccacgatg cgtccggcgt agaggatcga gatctcgatc ccgcgaaatt aatacgactc
181 actatagggg aattgtgagc ggataacaat tcccctctag aaataatttt gtttaacttt
241 aagaaggaga tataccatgg gttcttctca ccatcaccat caccatggtt cttctatgtc
301 ccctatacta ggttattgga aaattaaggg ccttgtgcaa cccactcgac ttcttttgga
361 atatcttgaa gaaaaatatg aagagcattt gtatgagcgc gatgaaggtg ataaatggcg
421 aaacaaaaag tttgaattgg gtttggagtt tcccaatctt ccttattata ttgatggtga
481 tgttaaatta acacagtcta tggccatcat acgttatata gctgacaagc acaacatgtt
541 gggtggttgt ccaaaagagc gtgcagagat ttcaatgctt gaaggagcgg ttttggatat
601 tagatacggt gtttcgagaa ttgcatatag taaagacttt gaaactctca aagttgattt
661 tcttagcaag ctacctgaaa tgctgaaaat gttcgaagat cgtttatgtc ataaaacata
721 tttaaatggt gatcatgtaa cccatcctga cttcatgttg tatgacgctc ttgatgttgt
781 tttatacatg gacccaatgt gcctggatgc gttcccaaaa ttagtttgtt ttaaaaaacg
841 tattgaagct atcccacaaa ttgataagta cttgaaatcc agcaagtata tagcatggcc
901 tttgcagggc tggcaagcca cgtttggtgg tggcgaccat cctccaaaag ggatcgagga
961 aaacctgtac ttccaatcca atattggaag tggataacgg atccgaattc gagcgccgtc
1021 gacaagcttg cggccgcact cgagcaccac caccaccacc actgagatcc ggctgctaac
1081 aaagcccgaa aggaagctga gttggctgct gccaccgctg agcaataact agcataaccc
1141 cttggggcct ctaaacgggt cttgaggggt tttttgctga aaggaggaac tatatccgga
1201 ttggcgaatg ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
1261 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
1321 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
1381 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
1441 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
1501 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
1561 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
1621 aacaaaaatt taacgcgaat tttaacaaac tagtaacgtt tacaatttca ggtggcactt
1681 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
1741 atccgctcat gaattaattc ttagaaaaac tcatcgagca tcaaatgaaa ctgcaattta
1801 ttcatatcag gattatcaat accatatttt tgaaaaagcc gtttctgtaa tgaaggagaa
1861 aactcaccga ggcagttcca taggatggca agatcctggt atcggtctgc gattccgact
1921 cgtccaacat caatacaacc tattaatttc ccctcgtcaa aaataaggtt atcaagtgag
1981 aaatcaccat gagtgacgac tgaatccggt gagaatggca aaagtttatg catttctttc
2041 cagacttgtt caacaggcca gccattacgc tcgtcatcaa aatcactcgc atcaaccaaa
2101 ccgttattca ttcgtgattg cgcctgagcg agacgaaata cgcgatcgct gttaaaagga
2161 caattacaaa caggaatcga atgcaaccgg cgcaggaaca ctgccagcgc atcaacaatg
2221 ttttcacctg aatcaggata ttcttctaat acctggaatg ctgttttccc ggggatcgca
2281 gtggtgagta accatgcatc atcaggagta cggataaaat gcttgatggt cggaagaggc
2341 ataaattccg tcagccagtt tagtctgacc atctcatctg taacatcatt ggcaacgcta
2401 cctttgccat gtttcagaaa caactctggc gcatcgggct tcccatacaa tcgatagatt
2461 gtcgcacctg attgcccgac attatcgcga gcccatttat acccatataa atcagcatcc
2521 atgttggaat ttaatcgcgg cctagagcaa gacgtttccc gttgaatatg gctcataaca
2581 ccccttgtat tactgtttat gtaagcagac agttttattg ttcatgacca aaatccctta
2641 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
2701 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
2761 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
2821 cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa
2881 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
2941 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
3001 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
3061 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
3121 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
3181 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
3241 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
3301 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
3361 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
3421 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg
3481 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc aatggtgcac tctcagtaca
3541 atctgctctg atgccgcata gttaagccag tatacactcc gctatcgcta cgtgactggg
3601 tcatggctgc gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc
3661 tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt
3721 tttcaccgtc atcaccgaaa cgcgcgaggc agctgcggta aagctcatca gcgtggtcgt
3781 gaagcgattc acagatgtct gcctgttcat ccgcgtccag ctcgttgagt ttctccagaa
3841 gcgttaatgt ctggcttctg ataaagcggg ccatgttaag ggcggttttt tcctgtttgg
3901 tcactgatgc ctccgtgtaa gggggatttc tgttcatggg ggtaatgata ccgatgaaac
3961 gagagaggat gctcacgata cgggttactg atgatgaaca tgcccggtta ctggaacgtt
4021 gtgagggtaa acaactggcg gtatggatgc ggcgggacca gagaaaaatc actcagggtc
4081 aatgccagcg cttcgttaat acagatgtag gtgttccaca gggtagccag cagcatcctg
4141 cgatgcagat ccggaacata atggtgcagg gcgctgactt ccgcgtttcc agactttacg
4201 aaacacggaa accgaagacc attcatgttg ttgctcaggt cgcagacgtt ttgcagcagc
4261 agtcgcttca cgttcgctcg cgtatcggtg attcattctg ctaaccagta aggcaacccc
4321 gccagcctag ccgggtcctc aacgacagga gcacgatcat gcgcacccgt ggggccgcca
4381 tgccggcgat aatggcctgc ttctcgccga aacgtttggt ggcgggacca gtgacgaagg
4441 cttgagcgag ggcgtgcaag attccgaata ccgcaagcga caggccgatc atcgtcgcgc
4501 tccagcgaaa gcggtcctcg ccgaaaatga cccagagcgc tgccggcacc tgtcctacga
4561 gttgcatgat aaagaagaca gtcataagtg cggcgacgat agtcatgccc cgcgcccacc
4621 ggaaggagct gactgggttg aaggctctca agggcatcgg tcgagatccc ggtgcctaat
4681 gagtgagcta acttacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
4741 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
4801 ggcgccaggg tggtttttct tttcaccagt gagacgggca acagctgatt gcccttcacc
4861 gcctggccct gagagagttg cagcaagcgg tccacgctgg tttgccccag caggcgaaaa
4921 tcctgtttga tggtggttaa cggcgggata taacatgagc tgtcttcggt atcgtcgtat
4981 cccactaccg agatatccgc accaacgcgc agcccggact cggtaatggc gcgcattgcg
5041 cccagcgcca tctgatcgtt ggcaaccagc atcgcagtgg gaacgatacc ctcattcagc
5101 atttgcatgg tttgttgaaa accggacatg gcactccagt cgccttcccg ttccgctatc
5161 ggctgaattt gattgcgagt gagatattta tgccagccag ccagacgcag acgcgccgag
5221 acagaactta atgggcccgc taacagcgcg atttgctggt gacccaatgc gaccagatgc
5281 tccacgccca gtcgcgtacc gtcttcatgg gagaaaataa tactgttgat gggtgtctgg
5341 tcagagacat caagaaataa cgccggaaca ttagtgcagg cagcttccac agcaatggca
5401 tcctggtcat ccagcggata gttaatgatc agcccactga cgcgttgcgc gagaagattg
5461 tgcaccgccg ctttacaggc ttcgacgccg cttcgttcta ccatcgacac caccacgctg
5521 gcacccagtt gatcggcgcg agatttaatc gccgcgacaa tttgcgacgg cgcgtgcagg
5581 gccagactgg aggtggcaac gccaatcagc aacgactgtt tgcccgccag ttgttgtgcc
5641 acgcggttgg gaatgtaatt cagctccgcc atcgccgctt ccactttttc ccgcgttttc
5701 gcagaaacgt ggctggcctg gttcaccacg cgggaaacgg tctgataaga gacaccggca
5761 tactctgcga catcgtataa cgttactggt ttcacattca ccaccctgaa ttgactctct
5821 tccgggcgct atcatgccat accgcgaaag gttttgcgcc attcgatggt gtccgggatc
5881 tcgacgctct cccttatgcg actcctgcat taggaagcag cccagtagta ggttgaggcc
5941 gttgagcacc gccgccgcaa ggaatggtgc atgcaaggag atggcgccca acagtccccc
6001 ggccacgggg cc
//
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