| 质粒名称: | pET-Bst-LF-Idaho |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 170469 |
| 存储实验室: | Jonathan Schmid-Burgk |
|
载体骨架基本信息 |
|
| 载体名称: | pET |
| 完整质粒大小: | 7080bp |
| 载体类型: | 细菌细胞表达 |
| 载体抗性: | Kanamycin, 50 μg/mL |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | Bst-LF-Idaho |
| 标签/融合蛋白: | N-terminal His-tag |
|
文献引用方法 |
|
| 材料和方法部分: | pET-Bst-LF-Idaho was a gift from Jonathan Schmid-Burgk (Addgene plasmid # 170469 ; http://n2t.net/addgene:170469 ; RRID:Addgene_170469) |
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
载体简介
载体序列
LOCUS pET-Bst-LF-Idaho 7080 bp ds-DNA circular SYN 23-JUL-2021
DEFINITION Expresses His-tagged Bst-LF-Idaho in E. coli.
ACCESSION .
VERSION .
KEYWORDS pET-Bst-LF-Idaho
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7080)
TITLE pET-BstLF-Idaho
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 7080)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Jul 23, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..7080
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 1..20
/label=T7
/note="T7 promoter, forward primer"
promoter 1..19
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 20..44
/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 59..81
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 94..111
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
CDS 121..144
/codon_start=1
/product="recognition and cleavage site for human
rhinovirus 3C and PreScission proteases"
/label=HRV 3C site
/translation="LEVLFQGP"
CDS 1945..1962
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
primer_bind complement(2015..2033)
/label=T7 Term
/note="T7 terminator, reverse primer"
terminator 2029..2076
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
rep_origin 2113..2568
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2200..2219)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 2410..2431
/label=F1ori-F
/note="F1 origin, forward primer"
CDS complement(2660..3475)
/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 3383..3402
/label=Kan-R
/note="Kanamycin resistance gene, reverse primer"
primer_bind 3467..3486
/label=pENTR-R
/note="pENTR vectors, reverse primer"
rep_origin 3597..4185
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 4086..4105
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 4339..4356
/label=L4440
/note="L4440 vector, forward primer"
misc_feature 4371..4510
/label=bom
/note="basis of mobility region from pBR322"
primer_bind complement(4436..4455)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 4596..4618
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
CDS complement(4612..4803)
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/label=rop
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
CDS complement(5612..6694)
/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 6656..6675
/label=LacI-R
/note="LacI, reverse primer"
promoter complement(6695..6772)
/gene="lacI"
/label=lacI promoter
primer_bind 6978..6997
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
ORIGIN
1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
61 tgtttaactt taagaaggag atataccatg aaacatcacc atcaccatca ctccgcgggt
121 ctggaagttc tgttccaggg gcccatggaa agtcccagca gcgaggaaga aaaaccgctg
181 gcgaaaatgg catttactct ggcggatcgc gttaccgaag aaatgctggc ggataaagcg
241 gccttggtag tggaagtggt ggaagaaaac tatcatgacg cgcctattgt tgggattgct
301 gttgttaacg agcacggacg cttctttctg cgcccggaaa ccgcgctggc cgatcctcaa
361 ttcgtggctt ggctgggaga tgagaccaaa aaaaaaagca tgttcgattc taaacgcgcg
421 gccgttgcgt taaaatggaa aggcatcgaa ctgtgtggtg tgagctttga tctcctgctg
481 gctgcctacc tcctcgatcc ggcccagggt gttgatgatg tggccgcagc ggccaaaatg
541 aagcaatatg aggccgtccg tcccgatgaa gctgtgtatg ggaagggcgc caaacgcgcg
601 gtacctgatg aaccagtcct ggcagagcac ctggtccgca aagcggcagc tatttgggaa
661 ctggaacgcc cctttctgga cgaattacgc cgtaatgaac aagatcgtct tctggtagaa
721 ctggagcagc cgctgtcgtc aattctcgct gaaatggaat ttgcgggtgt taaagttgac
781 acgaaacgcc tggagcagat gggtaaagaa ttggccgaac agctgggtac cgtcgaacaa
841 cgtatttatg agttagcggg acaggaattt aatatcaatt ccccgaagca gctgggggtg
901 attctctttg agaaattgca gctccctgtc cttaagaaaa ccaaaaccgg ttactcaacc
961 agtgctgatg tgctggagaa actggccccg taccatgaga tcgtggagaa tattctgcac
1021 taccggcagc tcggtaagtt gcaaagcaca tatatcgaag gcctgctgaa agtagttcgc
1081 ccggacacca aaaaagtcca cacaatcttt aaccaggcgc tgacccaaac cggacgcctg
1141 tcctcaaccg aaccgaactt gcagaacatc ccgatccgtc ttgaggaggg tcggaagatc
1201 cggcaggcat ttgtccctag cgagagcgat tggcttattt tcgcggcaga ctacagccaa
1261 atcgaattgc gtgtgttagc acacattgcg gaagacgata atttgatgga ggcgtttcgc
1321 cgcgatcttg atattcacac gaaaaccgcg atggatattt ttcaggtgag tgaagatgaa
1381 gtgactccaa acatgcgtcg tcaagcaaaa gcagtcaatt tcggcatcgt ctatggtatc
1441 agcgattacg gtcttgctca gaatcttaac atcagccgga aagaggccgc cgaatttatt
1501 gagcgttatt tcgagagctt tccgggcgtt aaacgctaca tggagaacat cgtgcaagaa
1561 gctaagcaga aggggtatgt gaccactctt ctgcatcgtc ggcgctactt accggatatt
1621 actagtcgta acttcaatgt gcgctcattt gccgaacgta tggcaatgaa taccccgatc
1681 caggggtccg cagcagatat tatcaagaaa gcaatgatcg atttgaacgc ccggcttaaa
1741 gaggaacgcc ttcaggcgca cctgctgctt caggtacatg atgagcttat cctggaagcc
1801 ccgaaggaag aaatggaacg tctgtgccgt cttgttcctg aggtcatgga acaagccgtg
1861 acactgcgcg tgccgttgaa agtggattac cactatggaa gtacttggta cgatgcgaaa
1921 taaaagcttg cggccgcact cgagcaccac caccaccacc actgagatcc ggctgctaac
1981 aaagcccgaa aggaagctga gttggctgct gccaccgctg agcaataact agcataaccc
2041 cttggggcct ctaaacgggt cttgaggggt tttttgctga aaggaggaac tatatccgga
2101 ttggcgaatg ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
2161 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
2221 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
2281 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
2341 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
2401 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
2461 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
2521 aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttag gtggcacttt
2581 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
2641 tccgctcatg aattaattct tagaaaaact catcgagcat caaatgaaac tgcaatttat
2701 tcatatcagg attatcaata ccatattttt gaaaaagccg tttctgtaat gaaggagaaa
2761 actcaccgag gcagttccat aggatggcaa gatcctggta tcggtctgcg attccgactc
2821 gtccaacatc aatacaacct attaatttcc cctcgtcaaa aataaggtta tcaagtgaga
2881 aatcaccatg agtgacgact gaatccggtg agaatggcaa aagtttatgc atttctttcc
2941 agacttgttc aacaggccag ccattacgct cgtcatcaaa atcactcgca tcaaccaaac
3001 cgttattcat tcgtgattgc gcctgagcga gacgaaatac gcgatcgctg ttaaaaggac
3061 aattacaaac aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca tcaacaatat
3121 tttcacctga atcaggatat tcttctaata cctggaatgc tgttttcccg gggatcgcag
3181 tggtgagtaa ccatgcatca tcaggagtac ggataaaatg cttgatggtc ggaagaggca
3241 taaattccgt cagccagttt agtctgacca tctcatctgt aacatcattg gcaacgctac
3301 ctttgccatg tttcagaaac aactctggcg catcgggctt cccatacaat cgatagattg
3361 tcgcacctga ttgcccgaca ttatcgcgag cccatttata cccatataaa tcagcatcca
3421 tgttggaatt taatcgcggc ctagagcaag acgtttcccg ttgaatatgg ctcataacac
3481 cccttgtatt actgtttatg taagcagaca gttttattgt tcatgaccaa aatcccttaa
3541 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
3601 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
3661 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
3721 agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag
3781 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
3841 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
3901 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
3961 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
4021 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
4081 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
4141 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
4201 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
4261 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
4321 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg
4381 tattttctcc ttacgcatct gtgcggtatt tcacaccgca atggtgcact ctcagtacaa
4441 tctgctctga tgccgcatag ttaagccagt atacactccg ctatcgctac gtgactgggt
4501 catggctgcg ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct
4561 cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt
4621 ttcaccgtca tcaccgaaac gcgcgaggca gctgcggtaa agctcatcag cgtggtcgtg
4681 aagcgattca cagatgtctg cctgttcatc cgcgtccagc tcgttgagtt tctccagaag
4741 cgttaatgtc tggcttctga taaagcgggc catgttaagg gcggtttttt cctgtttggt
4801 cactgatgcc tccgtgtaag ggggatttct gttcatgggg gtaatgatac cgatgaaacg
4861 agagaggatg ctcacgatac gggttactga tgatgaacat gcccggttac tggaacgttg
4921 tgagggtaaa caactggcgg tatggatgcg gcgggaccag agaaaaatca ctcagggtca
4981 atgccagcgc ttcgttaata cagatgtagg tgttccacag ggtagccagc agcatcctgc
5041 gatgcagatc cggaacataa tggtgcaggg cgctgacttc cgcgtttcca gactttacga
5101 aacacggaaa ccgaagacca ttcatgttgt tgctcaggtc gcagacgttt tgcagcagca
5161 gtcgcttcac gttcgctcgc gtatcggtga ttcattctgc taaccagtaa ggcaaccccg
5221 ccagcctagc cgggtcctca acgacaggag cacgatcatg cgcacccgtg gggccgccat
5281 gccggcgata atggcctgct tctcgccgaa acgtttggtg gcgggaccag tgacgaaggc
5341 ttgagcgagg gcgtgcaaga ttccgaatac cgcaagcgac aggccgatca tcgtcgcgct
5401 ccagcgaaag cggtcctcgc cgaaaatgac ccagagcgct gccggcacct gtcctacgag
5461 ttgcatgata aagaagacag tcataagtgc ggcgacgata gtcatgcccc gcgcccaccg
5521 gaaggagctg actgggttga aggctctcaa gggcatcggt cgagatcccg gtgcctaatg
5581 agtgagctaa cttacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
5641 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
5701 gcgccagggt ggtttttctt ttcaccagtg agacgggcaa cagctgattg cccttcaccg
5761 cctggccctg agagagttgc agcaagcggt ccacgctggt ttgccccagc aggcgaaaat
5821 cctgtttgat ggtggttaac ggcgggatat aacatgagct gtcttcggta tcgtcgtatc
5881 ccactaccga gatatccgca ccaacgcgca gcccggactc ggtaatggcg cgcattgcgc
5941 ccagcgccat ctgatcgttg gcaaccagca tcgcagtggg aacgatgccc tcattcagca
6001 tttgcatggt ttgttgaaaa ccggacatgg cactccagtc gccttcccgt tccgctatcg
6061 gctgaatttg attgcgagtg agatatttat gccagccagc cagacgcaga cgcgccgaga
6121 cagaacttaa tgggcccgct aacagcgcga tttgctggtg acccaatgcg accagatgct
6181 ccacgcccag tcgcgtaccg tcttcatggg agaaaataat actgttgatg ggtgtctggt
6241 cagagacatc aagaaataac gccggaacat tagtgcaggc agcttccaca gcaatggcat
6301 cctggtcatc cagcggatag ttaatgatca gcccactgac gcgttgcgcg agaagattgt
6361 gcaccgccgc tttacaggct tcgacgccgc ttcgttctac catcgacacc accacgctgg
6421 cacccagttg atcggcgcga gatttaatcg ccgcgacaat ttgcgacggc gcgtgcaggg
6481 ccagactgga ggtggcaacg ccaatcagca acgactgttt gcccgccagt tgttgtgcca
6541 cgcggttggg aatgtaattc agctccgcca tcgccgcttc cactttttcc cgcgttttcg
6601 cagaaacgtg gctggcctgg ttcaccacgc gggaaacggt ctgataagag acaccggcat
6661 actctgcgac atcgtataac gttactggtt tcacattcac caccctgaat tgactctctt
6721 ccgggcgcta tcatgccata ccgcgaaagg ttttgcgcca ttcgatggtg tccgggatct
6781 cgacgctctc ccttatgcga ctcctgcatt aggaagcagc ccagtagtag gttgaggccg
6841 ttgagcaccg ccgccgcaag gaatggtgca tgcaaggaga tggcgcccaa cagtcccccg
6901 gccacggggc ctgccaccat acccacgccg aaacaagcgc tcatgagccc gaagtggcga
6961 gcccgatctt ccccatcggt gatgtcggcg atataggcgc cagcaaccgc acctgtggcg
7021 ccggtgatgc cggccacgat gcgtccggcg tagaggatcg agatctcgat cccgcgaaat
//
