| 质粒名称: | Ppj220(JBEI-13483) |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 89786 |
| 存储实验室: | Harry Beller |
|
载体骨架基本信息 |
|
| 载体名称: | pET24 |
| 载体出品公司: | -- |
| 空载体大小: | -- |
| 完整质粒大小: | 6730 bp |
| 载体修饰: | -- |
| 载体类型: | 细菌表达 |
| 筛选标记: | -- |
| 载体抗性: | 卡那霉素，50μg/mL |
| 生长温度: | 37 ℃ |
| 宿主菌: | BL21(DE) |
| 拷贝数: | -- |
|
插入基因信息 |
|
| 插入基因名称: | 3607 |
| 物种来源: | -- |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子: | -- |
| 标签/融合蛋白: | StrepII（插入件上的C端子） |
| 克隆方法: | Gibson Cloning |
|
文献引用方法 |
|
| 材料和方法部分: |
Ppj220(JBEI-13483) was a gift from Harry Beller (Addgene plasmid # 89786 ; http://n2t.net/addgene:89786 ; RRID:Addgene_89786)
|
| 参考文献部分: | Investigation of Proposed Ladderane Biosynthetic Genes from Anammox Bacteria by Heterologous Expression in E. coli. Javidpour P, Deutsch S, Mutalik VK, Hillson NJ, Petzold CJ, Keasling JD, Beller HR. PLoS One. 2016 Mar 14;11(3):e0151087. doi: 10.1371/journal.pone.0151087. eCollection 2016. PONE-D-16-02052 [pii] PubMed 26975050 |
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 Ppj220(JBEI-13483) 6730 bp ds-DNA circular SYN 13-MAY-2021
DEFINITION for expression of C-StrepII-3607.
ACCESSION .
VERSION .
KEYWORDS Ppj220(JBEI-13483)
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6730)
AUTHORS Javidpour P, Deutsch S, Mutalik VK, Hillson NJ, Petzold CJ, Keasling
JD, Beller HR
TITLE Investigation of Proposed Ladderane Biosynthetic Genes from Anammox
Bacteria by Heterologous Expression in E. coli.
JOURNAL PLoS One. 2016 Mar 14;11(3):e0151087. doi:
10.1371/journal.pone.0151087. eCollection 2016.
PUBMED 26975050
REFERENCE 2 (bases 1 to 6730)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..6730
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 921..940
/label=LacI-R
/note="LacI, reverse primer"
promoter complement(960..1037)
/gene="lacI"
/label=lacI promoter
primer_bind 1243..1262
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
primer_bind 1346..1365
/label=T7
/note="T7 promoter, forward primer"
promoter 1346..1364
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 1365..1389
/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 1404..1426
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 2907..2930
/codon_start=1
/product="peptide that binds Strep-Tactin(R), an engineered
form of streptavidin"
/label=Strep-Tag II
/translation="WSHPQFEK"
CDS 2940..2957
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
primer_bind complement(3010..3028)
/label=T7 Term
/note="T7 terminator, reverse primer"
terminator 3024..3071
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
rep_origin 3108..3563
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(3195..3214)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 3405..3426
/label=F1ori-F
/note="F1 origin, forward primer"
CDS complement(3655..4470)
/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 4378..4397
/label=Kan-R
/note="Kanamycin resistance gene, reverse primer"
primer_bind 4462..4481
/label=pENTR-R
/note="pENTR vectors, reverse primer"
rep_origin 4592..5180
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 5081..5100
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 5334..5351
/label=L4440
/note="L4440 vector, forward primer"
misc_feature 5366..5505
/label=bom
/note="basis of mobility region from pBR322"
primer_bind complement(5431..5450)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 5591..5613
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
CDS complement(5607..5798)
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/label=rop
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
CDS complement(6607..959)
/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"
ORIGIN
1 ggcaacagct gattgccctt caccgcctgg ccctgagaga gttgcagcaa gcggtccacg
61 ctggtttgcc ccagcaggcg aaaatcctgt ttgatggtgg ttaacggcgg gatataacat
121 gagctgtctt cggtatcgtc gtatcccact accgagatat ccgcaccaac gcgcagcccg
181 gactcggtaa tggcgcgcat tgcgcccagc gccatctgat cgttggcaac cagcatcgca
241 gtgggaacga tgccctcatt cagcatttgc atggtttgtt gaaaaccgga catggcactc
301 cagtcgcctt cccgttccgc tatcggctga atttgattgc gagtgagata tttatgccag
361 ccagccagac gcagacgcgc cgagacagaa cttaatgggc ccgctaacag cgcgatttgc
421 tggtgaccca atgcgaccag atgctccacg cccagtcgcg taccgtcttc atgggagaaa
481 ataatactgt tgatgggtgt ctggtcagag acatcaagaa ataacgccgg aacattagtg
541 caggcagctt ccacagcaat ggcatcctgg tcatccagcg gatagttaat gatcagccca
601 ctgacgcgtt gcgcgagaag attgtgcacc gccgctttac aggcttcgac gccgcttcgt
661 tctaccatcg acaccaccac gctggcaccc agttgatcgg cgcgagattt aatcgccgcg
721 acaatttgcg acggcgcgtg cagggccaga ctggaggtgg caacgccaat cagcaacgac
781 tgtttgcccg ccagttgttg tgccacgcgg ttgggaatgt aattcagctc cgccatcgcc
841 gcttccactt tttcccgcgt tttcgcagaa acgtggctgg cctggttcac cacgcgggaa
901 acggtctgat aagagacacc ggcatactct gcgacatcgt ataacgttac tggtttcaca
961 ttcaccaccc tgaattgact ctcttccggg cgctatcatg ccataccgcg aaaggttttg
1021 cgccattcga tggtgtccgg gatctcgacg ctctccctta tgcgactcct gcattaggaa
1081 gcagcccagt agtaggttga ggccgttgag caccgccgcc gcaaggaatg gtgcatgcaa
1141 ggagatggcg cccaacagtc ccccggccac ggggcctgcc accataccca cgccgaaaca
1201 agcgctcatg agcccgaagt ggcgagcccg atcttcccca tcggtgatgt cggcgatata
1261 ggcgccagca accgcacctg tggcgccggt gatgccggcc acgatgcgtc cggcgtagag
1321 gatcgagatc tcgatcccgc gaaattaata cgactcacta taggggaatt gtgagcggat
1381 aacaattccc ctctagaaat aattttgttt aactttaaga aggagatata catatgagta
1441 gcaccaaacc aattaaacca catattcaaa gtaccaagat tgacacgctg tatccgagtt
1501 acgatgtgat tgttattggt gccggtatcg ccggtctgat ttgtgccgcc ttcctggcca
1561 aaagtggtaa gaaagccctg ctgattgagc aacatttcat tccaggtggc tactgtacca
1621 gctttaaacg taagggtttc ctgtttgatg cggcggttca tcatattggc ggttgcggta
1681 aatggagcgt ggtgggtcgt tgtttcaaaa tgctgggcat cgaaatggac ttttaccatc
1741 tggacccaat ggataatctg atcttcccaa cgtttagcat tgaaattcca gccgatctgg
1801 atgaatacct gctgcgcctg caaaagcgtt acccacatga aaaggataac attaagtggt
1861 tttttcaaga ctttatccgc ctgtaccgcg ccaccttcca taatgagaaa agtcaaatca
1921 tcgagaaata caaagatatg acgtacggca tgatgctgga tcatttcttt gatgatagtg
1981 aactgaaaat ggttctgagc ggccagtggg gttacgtggg tctggccccg agcaaggcca
2041 gcgccattgc catgagccaa atgatgatta actatctgaa agatggtgcc ttttttccaa
2101 agggtggtac ccaagaattt gccaatgcca tttttcgtaa gtttatcgat ttcggcggtc
2161 atgttatgct gagtagtaaa gtgaagaaaa tcctgaagaa gggcccaagt gttaatggtg
2221 tgaaactgaa tgacgataag gaaattcata gcagtgttgt ggtgagcaat atcgatgcgc
2281 gtcagacctt ttatcatctg ctgaacgacg gtaatgatgt ggatgccgtt tacctgaaaa
2341 acattgaggg tatgaaagaa agttgtagtt tttttctgct gtacctgggt atcggtaaag
2401 gcattgacct gaatacgctg aagcgcggct tctacctgag tggtagcaat ttcgacagca
2461 tcgacagtga atggatgtat attagtgtgc caaccagtat ttgtcaagcg ctgagtccac
2521 aaaataagca aattatcagt atcgtggtgt acaagaaaga agagttctac aaaaatgtga
2581 ccgattggga actgctgaaa cagaatattg tgagcagtac gctggagcgc ctggaaagtt
2641 acctgccgga tattaagaaa catattgaga tcattgaagt tgccacccca aaaaccctgg
2701 aacgctacac cctgaacacg aacggtgccg cgtatggttg ggccgtgagt gttgaccaaa
2761 tctgggaaca gcgtctgagc ccaacgacgc cgattagtgg tctgtatctg gcgggtcatt
2821 ggacgcgccc aggtccaggt atctgcgccg tggttagtag tggttggagt gtggccaacc
2881 tgattctgaa aaattggcgt agcgcttgga gccacccgca gttcgaaaaa taactcgagc
2941 accaccacca ccaccactga gatccggctg ctaacaaagc ccgaaaggaa gctgagttgg
3001 ctgctgccac cgctgagcaa taactagcat aaccccttgg ggcctctaaa cgggtcttga
3061 ggggtttttt gctgaaagga ggaactatat ccggattggc gaatgggacg cgccctgtag
3121 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
3181 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
3241 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
3301 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
3361 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
3421 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
3481 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa
3541 caaaatatta acgcttacaa tttaggtggc acttttcggg gaaatgtgcg cggaacccct
3601 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgaatta attcttagaa
3661 aaactcatcg agcatcaaat gaaactgcaa tttattcata tcaggattat caataccata
3721 tttttgaaaa agccgtttct gtaatgaagg agaaaactca ccgaggcagt tccataggat
3781 ggcaagatcc tggtatcggt ctgcgattcc gactcgtcca acatcaatac aacctattaa
3841 tttcccctcg tcaaaaataa ggttatcaag tgagaaatca ccatgagtga cgactgaatc
3901 cggtgagaat ggcaaaagtt tatgcatttc tttccagact tgttcaacag gccagccatt
3961 acgctcgtca tcaaaatcac tcgcatcaac caaaccgtta ttcattcgtg attgcgcctg
4021 agcgagacga aatacgcgat cgctgttaaa aggacaatta caaacaggaa tcgaatgcaa
4081 ccggcgcagg aacactgcca gcgcatcaac aatattttca cctgaatcag gatattcttc
4141 taatacctgg aatgctgttt tcccggggat cgcagtggtg agtaaccatg catcatcagg
4201 agtacggata aaatgcttga tggtcggaag aggcataaat tccgtcagcc agtttagtct
4261 gaccatctca tctgtaacat cattggcaac gctacctttg ccatgtttca gaaacaactc
4321 tggcgcatcg ggcttcccat acaatcgata gattgtcgca cctgattgcc cgacattatc
4381 gcgagcccat ttatacccat ataaatcagc atccatgttg gaatttaatc gcggcctaga
4441 gcaagacgtt tcccgttgaa tatggctcat aacacccctt gtattactgt ttatgtaagc
4501 agacagtttt attgttcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
4561 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
4621 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
4681 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc
4741 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
4801 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
4861 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
4921 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
4981 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
5041 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
5101 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
5161 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
5221 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
5281 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
5341 cagtgagcga ggaagcggaa gagcgcctga tgcggtattt tctccttacg catctgtgcg
5401 gtatttcaca ccgcaatggt gcactctcag tacaatctgc tctgatgccg catagttaag
5461 ccagtataca ctccgctatc gctacgtgac tgggtcatgg ctgcgccccg acacccgcca
5521 acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct
5581 gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg
5641 aggcagctgc ggtaaagctc atcagcgtgg tcgtgaagcg attcacagat gtctgcctgt
5701 tcatccgcgt ccagctcgtt gagtttctcc agaagcgtta atgtctggct tctgataaag
5761 cgggccatgt taagggcggt tttttcctgt ttggtcactg atgcctccgt gtaaggggga
5821 tttctgttca tgggggtaat gataccgatg aaacgagaga ggatgctcac gatacgggtt
5881 actgatgatg aacatgcccg gttactggaa cgttgtgagg gtaaacaact ggcggtatgg
5941 atgcggcggg accagagaaa aatcactcag ggtcaatgcc agcgcttcgt taatacagat
6001 gtaggtgttc cacagggtag ccagcagcat cctgcgatgc agatccggaa cataatggtg
6061 cagggcgctg acttccgcgt ttccagactt tacgaaacac ggaaaccgaa gaccattcat
6121 gttgttgctc aggtcgcaga cgttttgcag cagcagtcgc ttcacgttcg ctcgcgtatc
6181 ggtgattcat tctgctaacc agtaaggcaa ccccgccagc ctagccgggt cctcaacgac
6241 aggagcacga tcatgcgcac ccgtggggcc gccatgccgg cgataatggc ctgcttctcg
6301 ccgaaacgtt tggtggcggg accagtgacg aaggcttgag cgagggcgtg caagattccg
6361 aataccgcaa gcgacaggcc gatcatcgtc gcgctccagc gaaagcggtc ctcgccgaaa
6421 atgacccaga gcgctgccgg cacctgtcct acgagttgca tgataaagaa gacagtcata
6481 agtgcggcga cgatagtcat gccccgcgcc caccggaagg agctgactgg gttgaaggct
6541 ctcaagggca tcggtcgaga tcccggtgcc taatgagtga gctaacttac attaattgcg
6601 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
6661 ggccaacgcg cggggagagg cggtttgcgt attgggcgcc agggtggttt ttcttttcac
6721 cagtgagacg
//
