| 质粒名称: | pTOX8 | |
|---|---|---|
| 出品公司: | Addgene | |
| 目录编号: | 127453 | |
| 存储实验室: | Matthew Waldor | |
|
载体骨架基本信息 |
||
| 载体名称: | pDS132 | |
| 载体出品公司: | Clontech | |
| 空载体大小: | 4038 bp | |
| 完整质粒大小: | 6629bp | |
| 载体类型: | 细菌表达 | |
| 载体抗性: | Chloramphenicol, 25 μg/mL | |
| 生长温度: | 37 ℃ | |
| 宿主菌: | Pir1 | |
| 拷贝数: | 高拷贝 | |
|
插入基因信息 |
||
| 插入基因名称: | mqsR toxin | |
| 启动子: | pRha | |
| 克隆方法: | Gibson Cloning | |
| 5’测序引物 | tcatctttccctggttgcca | |
| 3’测序引物 |
tcaaacatgagaattcgcgga
(Common Sequencing Primers)
|
|
|
tsPurple chromoprotein | |
| 启动子: | apFAB46 | |
| 克隆方法: | Gibson Cloning | |
| 5’测序引物 | CGGTTTCTTACAGTGAAGGC | |
| 3’测序引物 |
AGATCCTTGGCGGCAAGAAA
(Common Sequencing Primers)
|
|
|
文献引用方法 |
||
| 材料和方法部分: | pTOX8 was a gift from Matthew Waldor (Addgene plasmid # 127453 ; http://n2t.net/addgene:127453 ; RRID:Addgene_127453) | |
| 参考文献部分: | A New Suite of Allelic Exchange Vectors for the Scarless Modification of Proteobacterial Genomes. Lazarus JE, Warr AR, Kuehl CJ, Giorgio RT, Davis BM, Waldor MK. Appl Environ Microbiol. 2019 Jun 14. pii: AEM.00990-19. doi: 10.1128/AEM.00990-19. 10.1128/AEM.00990-19 PubMed 31201277 | |
买家导航
pTOX8载体质粒基本信息
pTOX8质粒图谱载体图谱和pTOX8载体序列质粒序列多克隆位点信息
pTOX8质粒载体简介
pTOX8质粒序列载体序列
LOCUS pTOX8 6629 bp DNA circular SYN 25-AUG-2024
DEFINITION Encodes MqsR toxin and tsPurple chromoprotein. CAM-R.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6629)
AUTHORS Lazarus JE, Warr AR, Kuehl CJ, Giorgio RT, Davis BM, Waldor MK
TITLE A New Suite of Allelic Exchange Vectors for the Scarless
Modification of Proteobacterial Genomes.
JOURNAL Appl Environ Microbiol. 2019 Jun 14. pii: AEM.00990-19. doi:
10.1128/AEM.00990-19.
PUBMED 31201277
REFERENCE 2 (bases 1 to 6629)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 25, 2024 from SnapGene Server 7.0.3
https://www.biofeng.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Appl
Environ Microbiol. 2019 Jun 14. pii: AEM.00990-19. doi:
10.1128/AEM.00990-19."
FEATURES Location/Qualifiers
source 1..6629
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 326..428
/label=cat promoter
/note="promoter of the E. coli cat gene encoding
chloramphenicol acetyltransferase"
CDS 429..1088
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/label=CmR
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPELRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
primer_bind complement(495..514)
/label=CAT-R
/note="Chloramphenicol resistance gene, reverse primer"
terminator 1189..1275
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 1367..1394
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
regulatory 1448..1457
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
promoter 1597..1715
/label=rhaB promoter
/note="promoter of the E. coli rhaBAD operon, conferring
tight induction with L-rhamnose and repression with
D-glucose in the presence of RhaR and RhaS (Giacalone et
al., 2006)"
terminator 2095..2341
/label=lambda tL3 terminator
/note="transcription terminator tL3 from phage lambda"
protein_bind 2377..2393
/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)."
rep_origin 2428..2816
/label=R6K gamma ori
/note="gamma replication origin from E. coli plasmid R6K;
requires the R6K initiator protein pi for replication"
CDS complement(3434..3805)
/codon_start=1
/gene="traJ"
/product="oriT-recognizing protein"
/label=traJ
/translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
KQDELGKVMMGVVRPRAEP"
oriT 3838..3947
/note="incP origin of transfer"
primer_bind complement(4628..4647)
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
primer_bind complement(4786..4808)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
RBS 4972..4980
/label=Shine-Dalgarno sequence
/note="full consensus sequence for ribosome-binding sites
upstream of start codons in E. coli; complementary to a
region in the 3' end of the 16S rRNA (Chen et al., 1994)"
RBS 5902..5913
/note="strong bacterial ribosome binding site (Elowitz and
Leibler, 2000)"
ORIGIN
1 ttggcggatg agagggtacc gcatgagctt cgagctaatc gcgcaaaaaa ccccgcttcg
61 gcggggtttt ttcgctgatc acgtacgata tcttcgaacc ggtgcacatg gtgtacaggg
121 ccctaggata ggacgtctta aggtttaaac caggttaatt aatttaaatg catcccggga
181 cgtctcgagc tcgatcggac cgcggccgct agcacgtata ccaagtgtcc tgttttcttg
241 ccgccaagga tctgatggcg caggggatca agatctctgt tgataccggg aagccctggg
301 ccaacttttg gcgaaaatga gacgttgatc ggcacgtaag aggttccaac tttcaccata
361 atgaaataag atcactaccg ggcgtatttt ttgagttatc gagattttca ggagctaagg
421 aagctaaaat ggagaaaaaa atcactggat ataccaccgt tgatatatcc caatggcatc
481 gtaaagaaca ttttgaggca tttcagtcag ttgctcaatg tacctataac cagaccgttc
541 agctggatat tacggccttt ttaaagaccg taaagaaaaa taagcacaag ttttatccgg
601 cctttattca cattcttgcc cgcctgatga atgctcatcc ggaattacgt atggcaatga
661 aagacggtga gctggtgata tgggatagtg ttcacccttg ttacaccgtt ttccatgagc
721 aaactgaaac gttttcatcg ctctggagtg aataccacga cgatttccgg cagtttctac
781 acatatattc gcaagatgtg gcgtgttacg gtgaaaacct ggcctatttc cctaaagggt
841 ttattgagaa tatgtttttc gtctcagcca atccctgggt gagtttcacc agttttgatt
901 taaacgtggc caatatggac aacttcttcg cccccgtttt caccatgggc aaatattata
961 cgcaaggcga caaggtgctg atgccgctgg cgattcaggt tcatcatgcc gtttgtgatg
1021 gcttccatgt cggcagaatg cttaatgaat tacaacagta ctgcgatgag tggcagggcg
1081 gggcgtaatt tttttaaggc agttattgtc ttcaaattcc cgttgcatcg atggcccccg
1141 atggtagtgt ggggtctccc catgcgagag tagggaactg ccaggcatca aataaaacga
1201 aaggctcagt cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc
1261 ctgagtagga caaatccgcc gggagcggat ttgaacgttg cgaagcaacg gcccggaggg
1321 tggcgggcag gacgcccgcc ataaactgcc aggcatcaaa ttaagcagaa ggccatcctg
1381 acggatggcc tttttgcgtg gccagtgcca agcttgcatg cctgcagctg tccggaacta
1441 gtcgaggcca tggtggcctc ctgatgtcgt caacacggcg aaatagtaat cacgaggtca
1501 ggttcttacc ttaaattttc gacggaaaac cacgtaaaaa acgtcgattt ttcaagatac
1561 agcgtgaatt ttcaggaaat gcggtgagca tcacatcacc acaattcagc aaattgtgaa
1621 catcatcacg ttcatctttc cctggttgcc aatggcccat tttcctgtca gtaacgagaa
1681 ggtcgcgaat tcaggcgctt tttagactgg tcgtaatgaa attcagcagg atcaccatat
1741 ggaaaaacgc acaccacata cacgtttgag tcaggttaaa aaacttgtca atgccgggca
1801 agttcgtaca acacgtagtg ccctgttaaa tgcagatgag ttaggtttgg attttgatgg
1861 tatgtgtaat gttatcattg gattatcaga gagcgacttt tataaaagca tgaccaccta
1921 ctctgatcat actatctggc aggatgttta cagacccagg cttgttacag gccaggttta
1981 tcttaaaatt acggtaattc atgacgtact gatcgtctcg tttaaggaga agtaaggatc
2041 cgcgaattct catgtttgac agcttatcac tgatcagtga attaatggcg atgacgcatc
2101 ctcacgataa tatccgggta ggcgcaatca ctttcgtctc tactccgtta caaagcgagg
2161 ctgggtattt cccggccttt ctgttatccg aaatccactg aaagcacagc ggctggctga
2221 ggagataaat aataaacgag gggctgtatg cacaaagcat cttctgttga gttaagaacg
2281 agtatcgaga tggcacatag ccttgctcaa attggaatca ggtttgtgcc aataccagta
2341 gaaacagacg aagaagtcgt gggggaattc cacaaattgt tatccgctca caattccaca
2401 tgtggaattc cacatgtgga attcccatgt cagccgttaa gtgttcctgt gtcactcaaa
2461 attgctttga gaggctctaa gggcttctca gtgcgttaca tccctggctt gttgtccaca
2521 accgttaaac cttaaaagct ttaaaagcct tatatattct tttttttctt ataaaactta
2581 aaaccttaga ggctatttaa gttgctgatt tatattaatt ttattgttca aacatgagag
2641 cttagtacgt gaaacatgag agcttagtac gttagccatg agagcttagt acgttagcca
2701 tgagggttta gttcgttaaa catgagagct tagtacgtta aacatgagag cttagtacgt
2761 gaaacatgag agcttagtac gtactatcaa caggttgaac tgctgatctt cagatcctct
2821 acgccggacg catcgtggcc ggatccagcc gaccaggctt tccacgcccg cgtgccgctc
2881 catgtcgttc gcgcggttct cggaaacgcg ctgccgcgtt tcgtgattgt cacgctcaag
2941 cccgtagtcc cgttcgagcg tcgcgcagag gtcagcgagg gcgcggtagg cccgatacgg
3001 ctcatggatg gtgtttcggg tcgggtgaat cttgttgatg gcgatatgga tgtgcaggtt
3061 gtcggtgtcg tgatgcacgg cactgacgcg ctgatgctcg gcgaagccaa gcccagcgca
3121 gatgcggtcc tcaatcgcgc gcaacgtctc cgcgtcgggc ttctctcccg cgcggaagct
3181 aaccagcagg tgataggtct tgtcggcctc ggaacgggtg ttgccgtgct gggtcgccat
3241 cacctcggcc atgacagcgg gcagggtgtt tgcctcgcag ttcgtgacgc gcacgtgacc
3301 caggcgctcg gtcttgcctt gctcgtcggt gatgtacttc accagctccg cgaagtcgct
3361 cttcttgatg gagcgcatgg ggacgtgctt ggcaatcacg cgcacccccc ggccgtttta
3421 gcggctaaaa aagtcatggc tctgccctcg ggcggaccac gcccatcatg accttgccaa
3481 gctcgtcctg cttctcttcg atcttcgcca gcagggcgag gatcgtggca tcaccgaacc
3541 gcgccgtgcg cgggtcgtcg gtgagccaga gtttcagcag gccgcccagg cggcccaggt
3601 cgccattgat gcgggccagc tcgcggacgt gctcatagtc cacgacgccc gtgattttgt
3661 agccctggcc gacggccagc aggtaggccg acaggctcat gccggccgcc gccgcctttt
3721 cctcaatcgc tcttcgttcg tctggaaggc agtacacctt gataggtggg ctgcccttcc
3781 tggttggctt ggtttcatca gccatccgct tgccctcatc tgttacgccg gcggtagccg
3841 gccagcctcg cagagcagga ttcccgttga gcaccgccag gtgcgaataa gggacagtga
3901 agaaggaaca cccgctcgcg ggtgggccta cttcacctat cctgcccggc tgacgccgtt
3961 ggatacacca aggaaagtct acacgaaccc tttggcaaaa tcctgtatat cgtgcgaaaa
4021 aggatggata taccgaaaaa atcgctataa tgaccccgaa gcagggttat gcagcggaaa
4081 agcgctgctt ccctgctgtt ttgtggaata tctaccgact ggaaacaggc aaatgcagga
4141 aattactgaa ctgaggggac aggcgagaga cgatgccaaa gagctacacc gacgagctgg
4201 ccgagtgggt tgaatcccgc gcggccaaga agcgccggcg tgatgaggct gcggttgcgt
4261 tcctggcggt gagggcggat gtcgaggcgg cgttagcgtc cggctatgcg ctcgtcacca
4321 tttgggagca catgcgggaa acggggaagg tcaagttctc ctacgagacg ttccgctcgc
4381 acgccaggcg gcacatcaag gccaagcccg ccgatgtgcc cgcaccgcag gccaaggctg
4441 cggaacccgc gccggcaccc aagacgccgg agccacggcg gccgaagcag gggggcaagg
4501 ctgaaaagcc ggcccccgct gcggccccga ccggcttcac cttcaaccca acaccggaca
4561 aaaaggatcc tctacgccgg acgcatcgtg gccggcatca ccggcgccac aggtgcggtt
4621 gctggcgcct atatcgccga catcaccgat ggggaagatc gggctcgcca cttcgggctc
4681 atgagcgctt gtttcggcgt gggtatggtg gcaggccccg tggccggggg actgttgggc
4741 gccatctcct tgctgcctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca
4801 gctcccggag acggtcacag cttgtctgta agcggatgcc gtaagtaagg taaaggcctc
4861 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
4921 aataaccctg ataaatgctt caataatatt gaaaaaggag cttatcaccg ataaggaggt
4981 tttttaatga cggtgctgca ctcggttgac ttcttcccta gcggcaatgc cagcgttgcc
5041 attgagccgc gcctgcctca agccgacttc ccggagcacc accacgactt ccacgagatc
5101 gttatcgtgg agcacggtac cggcatccac gttttcaacg gccaaccgta cacgattacg
5161 ggcggtaccg tgtgcttcgt tcgtgatcac gaccgccact tatacgagca cacggacaac
5221 ttatgcttaa ccaacgtttt ataccgtagc cctgaccgct tccaattcct ggcgggctta
5281 aaccaactgt taccgcagga attagacggc caatacccta gccattggcg tgtgaatcat
5341 tcggtgctgc aacaagttcg ccaattagtg gcgcaaatgg agcaacaaga gggcgagaac
5401 gacctgccga gcacggcgag ccgtgaaatt ctgttcatgc agctgttact gctgttacgc
5461 aagtcgagcc tgcaagaaaa tttagagaat tcggcgagcc gcctgaatct gctgttagcg
5521 tggttagaag atcacttcgc ggacgaagtt aactgggatg cggttgccga ccagttcagc
5581 ctgagcttac gcaccctgca ccgccaactg aaacaacaga ccggcttaac cccgcaacgc
5641 tatttaaacc gtttacgctt aatgaaggcg cgccacttac tgcgccattc ggaagcgtcg
5701 gtgaccgata ttgcgtacca ctgtggcttt tcggatagca atcatttcag caccctgttc
5761 cgtcgcgaat tcaattggag ccctcgcgac atccgccaag gccgcgacgg tttcttacag
5821 tgaaggcctt ctagagtcga cctgcaaaaa agagtattga cttcgcatct ttttgtacct
5881 ataatagatt cattactaga gaaagaggag aaatactaga tggcgtccct tgtaaagaag
5941 gatatgtgtg ttaagatgac aatggaggga accgtcaacg ggtatcactt taagtgtgtt
6001 ggcgagggag aaggcaaacc gttcgagggt acacagaata tgcgcattcg tgtcaccgaa
6061 gggggccctt tgccctttgc tttcgatatt ttggccccgt gctgtatgta tggttcgaag
6121 acttttatca aacacgtaag cggaatccct gactacttca aggagagctt tcccgaaggc
6181 tttacttggg aacgtacgca gatttttgaa gacggaggag ttttgactgc ccatcaagat
6241 acatcacttg agggaaactg tttaatctac aaagttaagg tgttagggac aaattttcct
6301 gccaatggtc cggttatgca gaaaaagaca gcaggctggg aaccgtgtgt agagatgttg
6361 tacccccgcg acggggttct ttgcgggcaa tcccttatgg ccttgaaatg tactgatgga
6421 aatcacttga catcccattt acgcaccact tatcgctcgc gtaagccaag taatgcggtg
6481 aacatgccgg aatttcactt cggagaccat cgtattgaaa tcttgaaagc cgaacaagga
6541 aaattttatg aacaatacga atccgcagtg gcacgttatt cagatgttcc agaaaaggcg
6601 acttgataag gcatgcaagc ttggctgtt
//
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