| 質粒名稱: | pET-29b(+)_NSP4ss_SerM_NucA-His |
|---|---|
| 出品公司: | Addgene |
| 目錄編號: | 218190 |
| 存儲實驗室: | Florence Pojer |
|
載體骨架基本信息 |
|
| 載體名稱: | pET-29b(+) |
| 載體出品公司: | Novagen |
| 完整質粒大小: | 6081bp |
| 載體類型: | Bacterial Expression |
| 載體抗性: | Kanamycin, 50 μg/mL |
| 生長溫度: | 37 ℃ |
| 宿主菌: | T7 Express Iq lysY |
| 拷貝數: | 低拷貝 |
|
插入基因信息 |
|
| 插入基因名稱: | nucA |
| 物種來源: | Serratia marcescens |
| 基因突變: | codon optimized ecoli residues - 22-266 |
| GenBank編號 | AAA26560.1 P13717 |
| 啟動子: | T7 |
| 標簽/融合蛋白: |
NSP4 signal secretion peptide (N terminal on insert)
His (C terminal on insert)
|
| 克隆方法: | Gene Synthesis |
| 5’測序引物 | T7promoter |
| 3’測序引物 |
T7terminator
(Common Sequencing Primers)
|
|
文獻引用方法 |
|
| 材料和方法部分: | pET-29b(+)_NSP4ss_SerM_NucA-His was a gift from Florence Pojer (Addgene plasmid # 218190 ; //n2t.net/addgene:218190 ; RRID:Addgene_218190) |
| 參考文獻部分: | |
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pET-29b(+)_NSP4ss_SerM_NucA-His載體質粒基本信息
pET-29b(+)_NSP4ss_SerM_NucA-His質粒圖譜載體圖譜和pET-29b(+)_NSP4ss_SerM_NucA-His載體序列質粒序列多克隆位點信息
_NSP4ss_SerM_NucA-His質粒圖譜.png)
pET-29b(+)_NSP4ss_SerM_NucA-His質粒載體簡介
pET-29b(+)_NSP4ss_SerM_NucA-His質粒序列載體序列
LOCUS pET-29b(+)_NSP4ss_SerM_NucA-His 6081 bp DNA circular SYN 14-APR-2025
DEFINITION Bacterial expression of a periplasmically expressed His-tagged
Serratia Marcescens NucA (Benzonase).
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6081)
TITLE Homemade Benzonase
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 6081)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Apr 14, 2025 from SnapGene Server 8.0.1
//gt12365.cn
COMMENT SGRef: number: 1; type: "Journal Article"; journalName:
"Unpublished"
COMMENT Sequence Label: pET-29b(+)_NSP4ss_SerM_NucA-His
FEATURES Location/Qualifiers
source 1..6081
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind 929..948
/label=LacI-R
/note="LacI, reverse primer"
promoter complement(968..1045)
/gene="lacI"
/label=lacI promoter
primer_bind 1251..1270
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
primer_bind 1358..1377
/label=T7
/note="T7 promoter, forward primer"
promoter 1358..1376
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 1377..1401
/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 1416..1438
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 2253..2282
/codon_start=1
/product="10xHis affinity tag"
/label=10xHis
/translation="HHHHHHHHHH"
CDS 2295..2312
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
primer_bind complement(2365..2383)
/label=T7 Term
/note="T7 terminator, reverse primer"
terminator 2379..2426
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
rep_origin 2463..2918
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2550..2569)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 2760..2781
/label=F1ori-F
/note="F1 origin, forward primer"
CDS complement(3011..3826)
/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(3011..3826)
/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 3734..3753
/label=Kan-R
/note="Kanamycin resistance gene, reverse primer"
primer_bind 3818..3837
/label=pENTR-R
/note="pENTR vectors, reverse primer"
rep_origin 3948..4536
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 4437..4456
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 4690..4707
/label=L4440
/note="L4440 vector, forward primer"
misc_feature 4722..4864
/label=bom
/note="basis of mobility region from pBR322"
primer_bind complement(4790..4809)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 4950..4972
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
CDS complement(4966..5157)
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/label=rop
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
CDS complement(join(5966..6081,1..967))
/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 gtgagacggg caacagctga ttgcccttca ccgcctggcc ctgagagagt tgcagcaagc
61 ggtccacgct ggtttgcccc agcaggcgaa aatcctgttt gatggtggtt aacggcggga
121 tataacatga gctgtcttcg gtatcgtcgt atcccactac cgagatgtcc gcaccaacgc
181 gcagcccgga ctcggtaatg gcgcgcattg cgcccagcgc catctgatcg ttggcaacca
241 gcatcgcagt gggaacgatg ccctcattca gcatttgcat ggtttgttga aaaccggaca
301 tggcactcca gtcgccttcc cgttccgcta tcggctgaat ttgattgcga gtgagatatt
361 tatgccagcc agccagacgc agacgcgccg agacagaact taatgggccc gctaacagcg
421 cgatttgctg gtgacccaat gcgaccagat gctccacgcc cagtcgcgta ccgtcttcat
481 gggagaaaat aatactgttg atgggtgtct ggtcagagac atcaagaaat aacgccggaa
541 cattagtgca ggcagcttcc acagcaatgg catcctggtc atccagcgga tagttaatga
601 tcagcccact gacgcgttgc gcgagaagat tgtgcaccgc cgctttacag gcttcgacgc
661 cgcttcgttc taccatcgac accaccacgc tggcacccag ttgatcggcg cgagatttaa
721 tcgccgcgac aatttgcgac ggcgcgtgca gggccagact ggaggtggca acgccaatca
781 gcaacgactg tttgcccgcc agttgttgtg ccacgcggtt gggaatgtaa ttcagctccg
841 ccatcgccgc ttccactttt tcccgcgttt tcgcagaaac gtggctggcc tggttcacca
901 cgcgggaaac ggtctgataa gagacaccgg catactctgc gacatcgtat aacgttactg
961 gtttcacatt caccaccctg aattgactct cttccgggcg ctatcatgcc ataccgcgaa
1021 aggttttgcg ccattcgatg gtgtccggga tctcgacgct ctcccttatg cgactcctgc
1081 attaggaagc agcccagtag taggttgagg ccgttgagca ccgccgccgc aaggaatggt
1141 gcatgcaagg agatggcgcc caacagtccc ccggccacgg ggcctgccac catacccacg
1201 ccgaaacaag cgctcatgag cccgaagtgg cgagcccgat cttccccatc ggtgatgtcg
1261 gcgatatagg cgccagcaac cgcacctgtg gcgccggtga tgccggccac gatgcgtccg
1321 gcgtagagga tcgagatcga tctcgatccc gcgaaattaa tacgactcac tataggggaa
1381 ttgtgagcgg ataacaattc ccctctagaa ataattttgt ttaactttaa gaaggagata
1441 tacatatgaa aaaaattacg gcagccgcag ggttgcttct tctggctgct caaccagcaa
1501 tggcagacac gttagagtct attgacaact gcgcagtggg ttgtccaacg ggaggaagct
1561 cgaatgtctc gattgtgcgt catgcctata cattaaacaa caactcgact acaaagttcg
1621 ccaactgggt tgcttaccac atcactaagg atactcctgc cagcgggaaa acgcgtaact
1681 ggaaaactga cccagcattg aatcccgcag atactcttgc tccagcggac tatacggggg
1741 cgaatgctgc acttaaagta gaccgcggac atcaggctcc gttggcttcg ttggcaggcg
1801 taagcgactg ggagagtctt aattacctta gcaatatcac accgcagaaa agcgacttaa
1861 atcaaggtgc ctgggctcgt cttgaagacc aagaacgtaa actgattgac cgcgctgata
1921 tttcaagcgt ttatacggtg acagggcctt tgtatgagcg cgatatggga aaattaccag
1981 ggacacagaa ggcgcacacc atccccagcg cctattggaa agtgattttc attaataact
2041 ctccagctgt aaaccactat gcggcatttt tatttgacca gaacacgcct aaaggcgccg
2101 atttttgcca attccgcgtg actgttgatg aaatcgaaaa acgcaccggc ttaatcatct
2161 gggctggcct tcccgacgac gtacaagcat ccttaaaaag taagccggga gtacttcccg
2221 aattgatggg ttgtaagaat tctggtaacg gacaccatca tcatcatcat caccatcacc
2281 attaatagct cgagcaccac caccaccacc actgagatcc ggctgctaac aaagcccgaa
2341 aggaagctga gttggctgct gccaccgctg agcaataact agcataaccc cttggggcct
2401 ctaaacgggt cttgaggggt tttttgctga aaggaggaac tatatccgga ttggcgaatg
2461 ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
2521 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
2581 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
2641 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
2701 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
2761 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
2821 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
2881 taacgcgaat tttaacaaaa tattaacgtt tacaatttca ggtggcactt ttcggggaaa
2941 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
3001 gaattaattc ttagaaaaac tcatcgagca tcaaatgaaa ctgcaattta ttcatatcag
3061 gattatcaat accatatttt tgaaaaagcc gtttctgtaa tgaaggagaa aactcaccga
3121 ggcagttcca taggatggca agatcctggt atcggtctgc gattccgact cgtccaacat
3181 caatacaacc tattaatttc ccctcgtcaa aaataaggtt atcaagtgag aaatcaccat
3241 gagtgacgac tgaatccggt gagaatggca aaagtttatg catttctttc cagacttgtt
3301 caacaggcca gccattacgc tcgtcatcaa aatcactcgc atcaaccaaa ccgttattca
3361 ttcgtgattg cgcctgagcg agacgaaata cgcgatcgct gttaaaagga caattacaaa
3421 caggaatcga atgcaaccgg cgcaggaaca ctgccagcgc atcaacaata ttttcacctg
3481 aatcaggata ttcttctaat acctggaatg ctgttttccc ggggatcgca gtggtgagta
3541 accatgcatc atcaggagta cggataaaat gcttgatggt cggaagaggc ataaattccg
3601 tcagccagtt tagtctgacc atctcatctg taacatcatt ggcaacgcta cctttgccat
3661 gtttcagaaa caactctggc gcatcgggct tcccatacaa tcgatagatt gtcgcacctg
3721 attgcccgac attatcgcga gcccatttat acccatataa atcagcatcc atgttggaat
3781 ttaatcgcgg cctagagcaa gacgtttccc gttgaatatg gctcataaca ccccttgtat
3841 tactgtttat gtaagcagac agttttattg ttcatgacca aaatccctta acgtgagttt
3901 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
3961 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
4021 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
4081 ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta
4141 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
4201 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
4261 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
4321 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
4381 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
4441 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
4501 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
4561 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
4621 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
4681 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg gtattttctc
4741 cttacgcatc tgtgcggtat ttcacaccgc atatatggtg cactctcagt acaatctgct
4801 ctgatgccgc atagttaagc cagtatacac tccgctatcg ctacgtgact gggtcatggc
4861 tgcgccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc
4921 atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc
4981 gtcatcaccg aaacgcgcga ggcagctgcg gtaaagctca tcagcgtggt cgtgaagcga
5041 ttcacagatg tctgcctgtt catccgcgtc cagctcgttg agtttctcca gaagcgttaa
5101 tgtctggctt ctgataaagc gggccatgtt aagggcggtt ttttcctgtt tggtcactga
5161 tgcctccgtg taagggggat ttctgttcat gggggtaatg ataccgatga aacgagagag
5221 gatgctcacg atacgggtta ctgatgatga acatgcccgg ttactggaac gttgtgaggg
5281 taaacaactg gcggtatgga tgcggcggga ccagagaaaa atcactcagg gtcaatgcca
5341 gcgcttcgtt aatacagatg taggtgttcc acagggtagc cagcagcatc ctgcgatgca
5401 gatccggaac ataatggtgc agggcgctga cttccgcgtt tccagacttt acgaaacacg
5461 gaaaccgaag accattcatg ttgttgctca ggtcgcagac gttttgcagc agcagtcgct
5521 tcacgttcgc tcgcgtatcg gtgattcatt ctgctaacca gtaaggcaac cccgccagcc
5581 tagccgggtc ctcaacgaca ggagcacgat catgcgcacc cgtggggccg ccatgccggc
5641 gataatggcc tgcttctcgc cgaaacgttt ggtggcggga ccagtgacga aggcttgagc
5701 gagggcgtgc aagattccga ataccgcaag cgacaggccg atcatcgtcg cgctccagcg
5761 aaagcggtcc tcgccgaaaa tgacccagag cgctgccggc acctgtccta cgagttgcat
5821 gataaagaag acagtcataa gtgcggcgac gatagtcatg ccccgcgccc accggaagga
5881 gctgactggg ttgaaggctc tcaagggcat cggtcgagat cccggtgcct aatgagtgag
5941 ctaacttaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
6001 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgcca
6061 gggtggtttt tcttttcacc a
//
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