| 质粒名称: | pET His6 Sumo TEV LIC cloning vector (1S) |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 29659 |
| 存储实验室: | Scott Gradia |
|
载体骨架基本信息 |
|
| 载体名称: | pET |
| 载体抗性: | 卡那霉素 |
| 空载体大小: | 5661 bp |
| 载体类型: | 细菌表达 |
| 高拷贝/低拷贝: | 低拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 标签/融合蛋白: | His6 Sumo TEV（主干上的N终端） |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | LIC tag（克隆过程中销毁） |
| 3' 克隆位点: | LIC tag（克隆过程中销毁） |
| 5' 测序引物 | Sumo forward (5'ttattgaggctcacagagaac) |
| 3' 测序引物 | T7 reverse |
|
文献引用方法 |
|
| 材料和方法部分: | pET His6 Sumo TEV LIC cloning vector (1S) was a gift from Scott Gradia (Addgene plasmid # 29659 ; http://n2t.net/addgene:29659 ; RRID:Addgene_29659) |
买家导航
载体基本信息
载体图谱质粒图谱和多克隆位点信息
质粒图谱.png)
载体简介
载体序列
LOCUS pET His6 Sumo TEV LIC cloning vector (1S) 5658 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pET His6 Sumo TEV LIC cloning vector (1S)
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5658)
TITLE LIC N-terminal fusion vectors for E. coli expression
REFERENCE 2 (bases 1 to 5658)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..5658
/organism="synthetic DNA construct"
/mol_type="other DNA"
CDS 1241..1432
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/label=rop
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
primer_bind complement(1426..1448)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
misc_feature 1534..1673
/label=bom
/note="basis of mobility region from pBR322"
primer_bind 1589..1608
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind complement(1688..1705)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(1859..2447)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(1939..1958)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind complement(2558..2577)
/label=pENTR-R
/note="pENTR vectors, reverse primer"
CDS 2569..3384
/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 complement(2642..2661)
/label=Kan-R
/note="Kanamycin resistance gene, reverse primer"
rep_origin complement(3477..3932)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(3614..3635)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 3826..3845
/label=F1ori-R
/note="F1 origin, reverse primer"
terminator 3969..4016
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
primer_bind 4012..4030
/label=T7 Term
/note="T7 terminator, reverse primer"
CDS complement(4083..4100)
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
CDS complement(4166..4186)
/codon_start=1
/product="tobacco etch virus (TEV) protease recognition and
cleavage site"
/label=TEV site
/translation="ENLYFQS"
CDS complement(4196..4486)
/codon_start=1
/gene="S. cerevisiae SMT3 (truncated)"
/product="cleavable ubiquitin-like protein tag"
/label=SUMO
/translation="MSDSEVNQEAKPEVKPEVKPETHINLKVSDGSSEIFFKIKKTTPL
RRLMEAFAKRQGKEMDSLRFLYDGIRIQADQTPEDLDMEDNDIIEAHREQIG"
CDS complement(4505..4522)
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
RBS 4541..4563
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
protein_bind 4578..4602
/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)."
primer_bind complement(4602..4621)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(4603..4621)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(4705..4724)
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
promoter 4930..5007
/gene="lacI"
/label=lacI promoter
CDS 5008..432
/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 complement(5027..5046)
/label=LacI-R
/note="LacI, reverse primer"
ORIGIN
1 ggcgactgga gtgccatgtc cggttttcaa caaaccatgc aaatgctgaa tgagggtatc
61 gttcccactg cgatgctggt tgccaacgat cagatggcgc tgggcgcaat gcgcgccatt
121 accgagtccg ggctgcgcgt tggtgcggat atctcggtag tgggatacga cgataccgaa
181 gacagctcat gttatatccc gccgttaacc accatcaaac aggattttcg cctgctgggg
241 caaaccagcg tggaccgctt gctgcaactc tctcagggcc aggcggtgaa gggcaatcag
301 ctgttgcccg tctcactggt gaaaagaaaa accaccctgg cgcccaatac gcaaaccgcc
361 tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa
421 agcgggcagt gagcgcaacg caattaatgt aagttagctc actcattagg caccgggatc
481 tcgaccgatg cccttgagag ccttcaaccc agtcagctcc ttccggtggg cgcggggcat
541 gactatcgtc gccgcactta tgactgtctt ctttatcatg caactcgtag gacaggtgcc
601 ggcagcgctc tgggtcattt tcggcgagga ccgctttcgc tggagcgcga cgatgatcgg
661 cctgtcgctt gcggtattcg gaatcttgca cgccctcgct caagccttcg tcactggtcc
721 cgccaccaaa cgtttcggcg agaagcaggc cattatcgcc ggcatggcgg ccccacgggt
781 gcgcatgatc gtgctcctgt cgttgaggac ccggctaggc tggcggggtt gccttactgg
841 ttagcagaat gaatcaccga tacgcgagcg aacgtgaagc gactgctgct gcaaaacgtc
901 tgcgacctga gcaacaacat gaatggtctt cggtttccgt gtttcgtaaa gtctggaaac
961 gcggaagtca gcgccctgca ccattatgtt ccggatctgc atcgcaggat gctgctggct
1021 accctgtgga acacctacat ctgtattaac gaagcgctgg cattgaccct gagtgatttt
1081 tctctggtcc cgccgcatcc ataccgccag ttgtttaccc tcacaacgtt ccagtaaccg
1141 ggcatgttca tcatcagtaa cccgtatcgt gagcatcctc tctcgtttca tcggtatcat
1201 tacccccatg aacagaaatc ccccttacac ggaggcatca gtgaccaaac aggaaaaaac
1261 cgcccttaac atggcccgct ttatcagaag ccagacatta acgcttctgg agaaactcaa
1321 cgagctggac gcggatgaac aggcagacat ctgtgaatcg cttcacgacc acgctgatga
1381 gctttaccgc agctgcctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca
1441 gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca
1501 gggcgcgtca gcgggtgttg gcgggtgtcg gggcgcagcc atgacccagt cacgtagcga
1561 tagcggagtg tatactggct taactatgcg gcatcagagc agattgtact gagagtgcac
1621 cattgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgctctt
1681 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
1741 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
1801 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
1861 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
1921 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
1981 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
2041 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
2101 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
2161 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
2221 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
2281 actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct
2341 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
2401 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
2461 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
2521 tgaacaataa aactgtctgc ttacataaac agtaatacaa ggggtgttat gagccatatt
2581 caacgggaaa cgtcttgctc taggccgcga ttaaattcca acatggatgc tgatttatat
2641 gggtataaat gggctcgcga taatgtcggg caatcaggtg cgacaatcta tcgattgtat
2701 gggaagcccg atgcgccaga gttgtttctg aaacatggca aaggtagcgt tgccaatgat
2761 gttacagatg agatggtcag actaaactgg ctgacggaat ttatgcctct tccgaccatc
2821 aagcatttta tccgtactcc tgatgatgca tggttactca ccactgcgat ccccgggaaa
2881 acagcattcc aggtattaga agaatatcct gattcaggtg aaaacattgt tgatgcgctg
2941 gcagtgttcc tgcgccggtt gcattcgatt cctgtttgta attgtccttt taacagcgat
3001 cgcgtatttc gtctcgctca ggcgcaatca cgaatgaata acggtttggt tgatgcgagt
3061 gattttgatg acgagcgtaa tggctggcct gttgaacaag tctggaaaga aatgcataaa
3121 cttttgccat tctcaccgga ttcagtcgtc actcatggtg atttctcact tgataacctt
3181 atttttgacg aggggaaatt aataggttgt attgatgttg gacgagtcgg aatcgcagac
3241 cgataccagg atcttgccat cctatggaac tgcctcggtg agttttctcc ttcattacag
3301 aaacggcttt ttcaaaaata tggtattgat aatcctgata tgaataaatt gcagtttcat
3361 ttgatgctcg atgagttttt ctaagaatta attcatgagc ggatacatat ttgaatgtat
3421 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgaaat
3481 tgtaaacgtt actagtttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt
3541 taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg
3601 gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt
3661 caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc
3721 aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg
3781 atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa
3841 aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc
3901 cgccgcgctt aatgcgccgc tacagggcgc gtcccattcg ccaatccgga tatagttcct
3961 cctttcagca aaaaacccct caagacccgt ttagaggccc caaggggtta tgctagttat
4021 tgctcagcgg tggcagcagc caactcagct tcctttcggg ctttgttagc agccggatct
4081 cagtggtggt ggtggtggtg ctcgagtgcg gccgcaagct tgtcgacggc gctcgaattc
4141 ggatccgtta tccacttcca atattggatt ggaagtacag gttttcctcg atcccaccaa
4201 tctgttctct gtgagcctca ataatatcgt tatcctccat gtccaaatct tcaggggtct
4261 gatcagcttg aattctaata ccgtcgtaca agaatcttaa ggagtccatt tccttaccct
4321 gtcttttagc gaacgcttcc atcagccttc ttaaaggagt ggtctttttg atcttgaaga
4381 agatctctga agatccatcg gacaccttta aattgatgtg agtctcaggc ttgacttctg
4441 gcttgacctc tggcttagct tcttgattga cttctgagtc cgacatgcta gccatagaag
4501 aaccatggtg atggtgatgg tgagaagaac ccatggtata tctccttctt aaagttaaac
4561 aaaattattt ctagagggga attgttatcc gctcacaatt cccctatagt gagtcgtatt
4621 aatttcgcgg gatcgagatc tcgatcctct acgccggacg catcgtggcc ggcatcaccg
4681 gcgccacagg tgcggttgct ggcgcctata tcgccgacat caccgatggg gaagatcggg
4741 ctcgccactt cgggctcatg agcgcttgtt tcggcgtggg tatggtggca ggccccgtgg
4801 ccgggggact gttgggcgcc atctccttgc atgcaccatt ccttgcggcg gcggtgctca
4861 acggcctcaa cctactactg ggctgcttcc taatgcagga gtcgcataag ggagagcgtc
4921 gagatcccgg acaccatcga atggcgcaaa acctttcgcg gtatggcatg atagcgcccg
4981 gaagagagtc aattcagggt ggtgaatgtg aaaccagtaa cgttatacga tgtcgcagag
5041 tatgccggtg tctcttatca gaccgtttcc cgcgtggtga accaggccag ccacgtttct
5101 gcgaaaacgc gggaaaaagt ggaagcggcg atggcggagc tgaattacat tcccaaccgc
5161 gtggcacaac aactggcggg caaacagtcg ttgctgattg gcgttgccac ctccagtctg
5221 gccctgcacg cgccgtcgca aattgtcgcg gcgattaaat ctcgcgccga tcaactgggt
5281 gccagcgtgg tggtgtcgat ggtagaacga agcggcgtcg aagcctgtaa agcggcggtg
5341 cacaatcttc tcgcgcaacg cgtcagtggg ctgatcatta actatccgct ggatgaccag
5401 gatgccattg ctgtggaagc tgcctgcact aatgttccgg cgttatttct tgatgtctct
5461 gaccagacac ccatcaacag tattattttc tcccatgaag acggtacgcg actgggcgtg
5521 gagcatctgg tcgcattggg tcaccagcaa atcgcgctgt tagcgggccc attaagttct
5581 gtctcggcgc gtctgcgtct ggctggctgg cataaatatc tcactcgcaa tcaaattcag
5641 ccgatagcgg aacgggaa
//
