| 质粒名称: | pHoss1 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 63158 |
| 存储实验室: | Attila Karsi |
|
载体骨架基本信息 |
|
| 载体名称: | pMAD |
| 载体出品公司: | Arnaud et al. 2004. Applied and Environmental Microbiology, 70:6887-6891 |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 9666 bp |
| 完整质粒大小: | 8995 bp |
| 载体修饰: | 利用BglII和BseRI限制酶从pMAD中去除耐热半乳糖苷酶（bgaB）。 |
| 载体类型: | 革兰氏阳性菌自杀质粒 |
| 筛选标记: | 红霉素 |
| 高拷贝/低拷贝: | -- |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | anti-secY-Pxyl/tetO-tetR |
| 物种来源: | -- |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | -- |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | NcoI(未破坏) |
| 3' 克隆位点: | BglII（克隆过程中被破坏） |
| 5' 测序引物 | GATCTAATGATTCAAACCCTTGTG |
| 3' 测序引物 | TGAAGTTACCATCACGGAAAAAGG |
|
文献引用方法 |
|
| 材料和方法部分: | pHoss1 was a gift from Attila Karsi (Addgene plasmid # 63158 ; http://n2t.net/addgene:63158 ; RRID:Addgene_63158) |
| 参考文献部分: | A novel suicide plasmid for efficient gene mutation in Listeria monocytogenes. Abdelhamed H, Lawrence ML, Karsi A. Plasmid. 2015 May 31;81:1-8. doi: 10.1016/j.plasmid.2015.05.003. 10.1016/j.plasmid.2015.05.003 PubMed 26038185 |
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 pHoss1 8859 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Suicide plasmid for efficient gene mutation in Gram-positive
bacteria.
ACCESSION .
VERSION .
KEYWORDS pHoss1
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8859)
AUTHORS Abdelhamed H, Lawrence ML, Karsi A
TITLE A novel suicide plasmid for efficient gene mutation in Listeria
monocytogenes.
JOURNAL Plasmid. 2015 May 31;81:1-8. doi: 10.1016/j.plasmid.2015.05.003.
PUBMED 26038185
REFERENCE 2 (bases 1 to 8859)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..8859
/organism="synthetic DNA construct"
/mol_type="other DNA"
CDS complement(4069..4692)
/codon_start=1
/gene="tetR from transposon Tn10"
/product="tetracycline repressor TetR"
/label=TetR
/note="TetR binds to the tetracycline operator tetO to
inhibit transcription. This inhibition can be relieved by
adding tetracycline or doxycycline."
/translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
KNKRALLDALAIEMLDRRHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
primer_bind 4600..4619
/label=Tet-R
/note="Tetracycline resistance gene, reverse primer"
protein_bind 4717..4735
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O1 for the tetR and tetA genes"
protein_bind 4802..4820
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O1 for the tetR and tetA genes"
CDS 6416..6607
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/label=rop
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
primer_bind complement(6601..6623)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
misc_feature 6709..6849
/label=bom
/note="basis of mobility region from pBR322"
primer_bind 6764..6783
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind complement(6864..6881)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(7035..7623)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(7115..7134)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(7794..8654)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind 8417..8436
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(8655..8759)
/gene="bla"
/label=AmpR promoter
primer_bind 8827..8845
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
ORIGIN
1 gaattaattc tcatgtttga cagcttatca tcgattcaca aaaaataggc acacgaaaaa
61 caagttaagg gatgcagttt atgcatccct taacttactt attaaataat ttatagctat
121 tgaaaagaga taagaattgt tcaaagctaa tattgtttaa atcgtcaatt cctgcatgtt
181 ttaaggaatt gttaaattga ttttttgtaa atattttctt gtattctttg ttaacccatt
241 tcataacgaa ataattatac ttttgtttat ctttgtgtga tattcttgat ttttttctac
301 ttaatctgat aagtgagcta ttcactttag gtttaggatg aaaatattct cttggaacca
361 tacttaatat agaaatatca acttctgcca ttaaaagtaa tgccaatgag cgttttgtat
421 ttaataatct tttagcaaac ccgtattcca cgattaaata aatctcatta gctatactat
481 caaaaacaat tttgcgtatt atatccgtac ttatgttata aggtatatta ccatatattt
541 tataggattg gtttttagga aatttaaact gcaatatatc cttgtttaaa acttggaaat
601 tatcgtgatc aacaagttta ttttctgtag ttttgcataa tttatggtct atttcaatgg
661 cagttacgaa attacacctc tttactaatt caagggtaaa atggcctttt cctgagccga
721 tttcaaagat attatcatgt tcatttaatc ttatatttgt cattatttta tctatattat
781 gttttgaagt aataaagttt tgactgtgtt ttatattttt ctcgttcatt ataaccctct
841 ttaatttggt tatatgaatt ttgcttatta acgattcatt ataaccactt attttttgtt
901 tggttgataa tgaactgtgc tgattacaaa aatactaaaa atgcccatat tttttcctcc
961 ttataaaatt agtataatta tagcacgagc tctgataaat atgaacatga tgagtgatcg
1021 ttaaatttat actgcaatcg gatgcgatta ttgaataaaa gatatgagag atttatctaa
1081 tttctttttt cttgtaaaaa aagaaagttc ttaaaggttt tatagttttg gtcgtagagc
1141 acacggttta acgacttaat tacgaagtaa ataagtctag tgtgttagac tttatgaaat
1201 ctatatacgt ttatatatat ttattatccg gaggtgtagc atgtctcatt caattttgag
1261 ggttgccaga gttaaaggat caagtaatac aaacgggata caaagacata atcaaagaga
1321 gaataaaaac tataataata aagacataaa tcatgaggaa acatataaaa attatgattt
1381 gattaacgca caaaatataa agtataaaga taaaattgat gaaacgattg atgagaatta
1441 ttcagggaaa cgtaaaattc ggtcagatgc aattcgacat gtggacggac tggttacaag
1501 tgataaagat ttctttgatg atttaagcgg agaagaaata gaacgatttt ttaaagatag
1561 cttggagttt ctagaaaatg aatacggtaa ggaaaatatg ctgtatgcga ctgtccatct
1621 ggatgaaaga gtcccacata tgcactttgg ttttgtccct ttaacagagg acgggagatt
1681 gtctgcaaaa gaacagttag gcaacaagaa agactttact caattacaag atagatttaa
1741 tgagtatgtg aatgagaaag gttatgaact tgaaagaggc acgtccaaag aggttacaga
1801 acgagaacat aaagcgatgg atcagtacaa gaaagatact gtatttcata aacaggaact
1861 gcaagaagtt aaggatgagt tacagaaggc aaataagcag ttacagagtg gaatagagca
1921 tatgaggtct acgaaaccct ttgattatga aaatgagcgt acaggtttgt tctctggacg
1981 tgaagagact ggtagaaaga tattaactgc tgatgaattt gaacgcctgc aagaaacaat
2041 ctcttctgca gaacggattg ttgatgatta cgaaaatatt aagagcacag actattacac
2101 agaaaatcaa gaattaaaaa aacgtagaga gagtttgaaa gaagtagtga atacatggaa
2161 agaggggtat cacgaaaaaa gtaaagaggt taataaatta aagcgagaga atgatagttt
2221 gaatgagcag ttgaatgtat cagagaaatt tcaagctagt acagtgactt tatatcgtgc
2281 tgcgagggcg aatttccctg ggtttgagaa agggtttaat aggcttaaag agaaattctt
2341 taatgattcc aaatttgagc gtgtgggaca gtttatggat gttgtacagg ataatgtcca
2401 gaaggtcgat agaaagcgtg agaaacagcg tacagacgat ttagagatgt agaggtactt
2461 ttatgccgag aaaacttttt gcgtgtgaca gtccttaaaa tatacttaga gcgtaagcga
2521 aagtagtagc gacagctatt aactttcggt tgcaaagctc taggattttt aatggacgca
2581 gcgcatcaca cgcaaaaagg aaattggaat aaatgcgaaa tttgagatgt taattaaaga
2641 cctttttgag gtcttttttt cttagatttt tggggttatt taggggagaa aacatagggg
2701 ggtactacga cctcccccct aggtgtccat tgtccattgt ccaaacaaat aaataaatat
2761 tgggttttta atgttaaaag gttgtttttt atgttaaagt gaaaaaaaca gatgttggga
2821 ggtacagtga tagttgtaga tagaaaagaa gagaaaaaag ttgctgttac tttaagactt
2881 acaacagaag aaaatgagat attaaataga atcaaagaaa aatataatat tagcaaatca
2941 gatgcaaccg gtattctaat aaaaaaatat gcaaaggagg aatacggtgc attttaaaca
3001 aaaaaagata gacagcactg gcatgctgcc tatctatgac taaattttgt taagtgtatt
3061 agcaccgtta ttatatcatg agcgaaaatg taataaaaga aactgaaaac aagaaaaatt
3121 caagaggacg taattggaca tttgttttat atccagaatc agcaaaagcc gagtggttag
3181 agtatttaaa agagttacac attcaatttg tagtgtctcc attacatgat agggatactg
3241 atacagaagg taggatgaaa aaagagcatt atcatattct agtgatgtat gagggtaata
3301 aatcttatga acagataaaa ataattacag aagaattgaa tgcgactatt ccgcagattg
3361 caggaagtgt gaaaggtctt gtgagatata tgcttcacat ggacgatcct aataaattta
3421 aatatcaaaa agaagatatg atagtttatg gcggtgtaga tgttgatgaa ttattaaaga
3481 aaacaacaac agatagatat aaattaatta aagaaatgat tgagtttatt gatgaacaag
3541 gaatcgtaga atttaagagt ttaatggatt atgcaatgaa gtttaaattt gatgattggt
3601 tcccgctttt atgtgataac tcggcgtatg ttattcaaga atatataaaa tcaaatcggt
3661 ataaatctga ccgatagatt ttgaatttaa gagtgtcaca agacactctt ttttcgcacc
3721 aacgaaaact ggtttaagcc gactgcgcaa aagacataat cgataagctt taatgcggta
3781 gtttatcaca gttaaattgc taacgcagtc aggcaccgtg tatgaaatct aacaatgcgc
3841 tcatcgtcat cctcggcacc gtcaccctgg atgctgtagg cataggcttg gttatgccgg
3901 tactgccggg cctcttgcgg gatcgaggcc tttagcatat tatgttgcca actgtcggaa
3961 cgagacttct ctaaaccttc ccggcttcat catgctctct taacactgct acatcaaccc
4021 cttgaatcga tcttgaagtt accatcacgg aaaaaggtta tgctgctttt aagacccact
4081 ttcacattta agttgttttt ctaatccgca tatgatcaat tcaaggccga ataagaaggc
4141 tggctctgca ccttggtgat caaataattc gatagcttgt cgtaataatg gcggcatact
4201 atcagtagta ggtgtttccc tttcttcttt agcgacttga tgctcttgat cttccaatac
4261 gcaacctaaa gtaaaatgcc ccacagcgct gagtgcatat aatgcattct ctagtgaaaa
4321 accttgttgg cataaaaagg ctaattgatt ttcgagagtt tcatactgtt tttctgtagg
4381 ccgtgtacct aaatgtactt ttgctccatc gcgatgactt agtaaagcac atctaaaact
4441 tttagcgtta ttacgtaaaa aatcttgcca gctttcccct tctaaagggc aaaagtgagt
4501 atggcgccta tctaacatct caatggctaa ggcgtcgagc aaagcccgct tattttttac
4561 atgccaatac aatgtaggct gctctacacc tagcttctgg gcgagtttac gggttgttaa
4621 accttcgatt ccgacctcat taagcagctc taatgcgctg ttaatcactt tacttttatc
4681 taatctagac atcattaatt cctccttttt gttgacactc tatcattgat agagttattt
4741 gtcaaactag ttttttattt ggatcccctc gagttcatga aaaactaaaa aaaatattga
4801 cactctatca ttgatagagt ataattaaaa taagcttgat gggaatatga taatagaaat
4861 accattacca acaccgaact gagtgatttg atcaccaagc catattaaga aagcagttcc
4921 tgctgtcaaa actagtgcta ttaataaata actcataatt gactgattga taatcagcgc
4981 acctttgaga taattattaa attggaatgc catacctata gattggataa atgctaaaga
5041 aattgctaaa taacgagtaa cgttatttaa ctttcttcta cctacttcac cttgttttgc
5101 ccattctgag aatttaggga caatatccat ttgtaataat tgcattacga ttgatgcagt
5161 gatgtagggt acaataccca ttgcaaaaat agaaaatcgt ttcaaggctc cgccaccaaa
5221 agtatttaat aactcagtgg caccttgaga accttgggga ttatcaaaag ctgcaggatt
5281 tactcctgga gctggtatat aagtccctat tttaaaaatt actaacattg ctagtgtgaa
5341 gaaaatcttg ttacgaacct cttttgttct aaagaagttc acaagggttt gaatcattag
5401 atcccatggt acccgggagc tcgaattcta gaagcttctg cagacgcgtc gacgtcatat
5461 ggatccgata tcgcccgacg cgaggctgga tggccttccc cattatgatt cttctcgctt
5521 ccggcggcat cgggatgccc gcgttgcagg ccatgctgtc caggcaggta gatgacgacc
5581 atcagggaca gcttcaagga tcgctcgcgg ctcttaccag cctaacttcg atcattggac
5641 cgctgatcgt cacggcgatt tatgccgcct cggcgagcac atggaacggg ttggcatgga
5701 ttgtaggcgc cgccctatac cttgtctgcc tccccgcgtt gcgtcgcggt gcatggagcc
5761 gggccacctc gacctgaatg gaagccggcg gcacctcgct aacggattca ccactccaag
5821 aattggagcc aatcaattct tgcggagaac tgtgaatgcg caaaccaacc cttggcagaa
5881 catatccatc gcgtccgcca tctccagcag ccgcacgcgg cgcatctcgg gcagcgttgg
5941 gtcctggcca cgggtgcgca tgatcgtgct cctgtcgttg aggacccggc taggctggcg
6001 gggttgcctt actggttagc agaatgaatc accgatacgc gagcgaacgt gaagcgactg
6061 ctgctgcaaa acgtctgcga cctgagcaac aacatgaatg gtcttcggtt tccgtgtttc
6121 gtaaagtctg gaaacgcgga agtcagcgcc ctgcaccatt atgttccgga tctgcatcgc
6181 aggatgctgc tggctaccct gtggaacacc tacatctgta ttaacgaagc gctggcattg
6241 accctgagtg atttttctct ggtcccgccg catccatacc gccagttgtt taccctcaca
6301 acgttccagt aaccgggcat gttcatcatc agtaacccgt atcgtgagca tcctctctcg
6361 tttcatcggt atcattaccc ccatgaacag aaatccccct tacacggagg catcagtgac
6421 caaacaggaa aaaaccgccc ttaacatggc ccgctttatc agaagccaga cattaacgct
6481 tctggagaaa ctcaacgagc tggacgcgga tgaacaggca gacatctgtg aatcgcttca
6541 cgaccacgct gatgagcttt accgcagctg cctcgcgcgt ttcggtgatg acggtgaaaa
6601 cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag
6661 cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggcg cagccatgac
6721 ccagtcacgt agcgatagcg gagtgtatac tggcttaact atgcggcatc agagcagatt
6781 gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac
6841 cgcatcaggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
6901 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
6961 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
7021 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
7081 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
7141 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
7201 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
7261 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
7321 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
7381 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
7441 ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg
7501 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
7561 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
7621 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
7681 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
7741 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
7801 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
7861 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
7921 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
7981 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
8041 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
8101 gccattgctg caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
8161 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
8221 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
8281 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
8341 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
8401 ccggcgtcaa cacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
8461 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
8521 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
8581 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
8641 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
8701 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
8761 acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc
8821 tataaaaata ggcgtatcac gaggcccttt cgtcttcaa
//
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