| 质粒名称: | pJB38-NWMN2930 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 84457 |
| 存储实验室: | Reindert Nijland |
|
载体骨架基本信息 |
|
| 载体名称: | pJB38 |
| 空载体大小: | 7015 bp |
| 完整质粒大小: | 8815 bp |
| 筛选标记: | also contains Chloramphenicol resistance gene, but only functional in Gram-postive bacteria. |
| 载体抗性: | 氨苄霉素 |
| 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: | Staphylococcus aureus NWMN genomic region NWMN29-30 |
| 物种来源: | Staphylocccus aureus |
| GenBank编号 | NC_009641.1 |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | EcoRI未破坏) |
| 3’克隆位点: | KpnI (未破坏) |
| 5’测序引物 | AACCTATAAAAATAGGCGTATCA |
|
文献引用方法 |
|
| 材料和方法部分: | pJB38-NWMN2930 was a gift from Reindert Nijland (Addgene plasmid # 84457 ; http://n2t.net/addgene:84457 ; RRID:Addgene_84457) |
| 参考文献部分: | Fluorescent reporters for markerless genomic integration in Staphylococcus aureus. de Jong NW, van der Horst T, van Strijp JA, Nijland R. Sci Rep. 2017 Mar 7;7:43889. doi: 10.1038/srep43889. 10.1038/srep43889 PubMed 28266573 |
买家导航
pJB38-NWMN2930载体质粒基本信息
pJB38-NWMN2930质粒图谱载体图谱和pJB38-NWMN2930载体序列质粒序列多克隆位点信息
pJB38-NWMN2930质粒载体简介
pJB38-NWMN2930质粒序列载体序列
LOCUS pJB38-NWMN2930 8815 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION pJB38 plasmid containing the S.aureus Newman genetic region between
genes 29 and 30.
ACCESSION .
VERSION .
KEYWORDS pJB38-NWMN2930
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8815)
AUTHORS de Jong NW, van der Horst T, van Strijp JA, Nijland R
TITLE Fluorescent reporters for markerless genomic integration in
Staphylococcus aureus.
JOURNAL Sci Rep. 2017 Mar 7;7:43889. doi: 10.1038/srep43889.
PUBMED 28266573
REFERENCE 2 (bases 1 to 8815)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..8815
/organism="synthetic DNA construct"
/mol_type="other DNA"
protein_bind 3811..3829
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O1 for the tetR and tetA genes"
protein_bind 3900..3918
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O1 for the tetR and tetA genes"
CDS 3943..4566
/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
KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
primer_bind complement(4016..4035)
/label=Tet-R
/note="Tetracycline resistance gene, reverse primer"
primer_bind complement(6557..6579)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
misc_feature 6665..6805
/label=bom
/note="basis of mobility region from pBR322"
primer_bind 6720..6739
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind complement(6820..6837)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(6991..7579)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(7071..7090)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(7750..8610)
/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 8373..8392
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(8611..8715)
/gene="bla"
/label=AmpR promoter
primer_bind 8783..8801
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
ORIGIN
1 gaattcagtg gctacattcg aacatatcaa agcatacgat agaaaaacaa taagttatga
61 tatgttacat gtaacaccac ctatgggtcc cttagatgta gtaaaagaaa gtacactttc
121 agatagtgag ggttgggtag atgttaaccc aaccacatta cagcataaaa gctactctaa
181 tgtatttgca cttggtgatg cttcaaatgt acctacttca aaaacaggcg cagctattcg
241 taagcaagca cctatcgtcg ctaataattt attgcaagtg atgaataatc aaatgttaac
301 gcatcattat gatggttata cttcatgccc tattgttact ggatataata ggttaatact
361 tgcagagttt gattataata aaaatactaa agaaacaatg ccgtttaatc aggccaaaga
421 acgtagaagt atgtatatat ttaagaaaga tttattacct aaaatgtatt ggtacggcat
481 gctaaaagga ttaatataat aaagtacaga aaacaataaa tttttaatga aaaatctttt
541 actataaaag attaagtatt taaatgacgt gtcagtgttg tgtttatatg tcgtgaattt
601 ttagctctaa atagtataag attgaaaaag ttgttactgt tttaaatgat cacgatgaag
661 tcattcaata agaatgatta tgaaaataga aacagcagta agatattttc taattgaaaa
721 tcatctcact gctgtttttt aaaggtttat acctcatcct ctaaattatt taaaaataat
781 taatggtatt tgagcacgtt tagcgacttt atgactgaca ttaccaattt ccatttcttg
841 ccagatattc aaaccacgtg tactcaaaat gatagcttgg tatgtacctc caatagtaat
901 ttcaataact ttgtctgttg aacactaaga gcaattttaa tttcataatg tgttgtaaac
961 attttttttg attggagttt ttttctgagt taaacgatat cctgatgtat ttttaatttt
1021 gcaccatttc caaaaggata agtgacataa gtaaaaaggc atcatcggga gttatcctat
1081 caggaaaacc aagataatac ctaagtagaa agtgttcaat ccgtgttaaa ttgggaaata
1141 tcatccataa actttattac tcatactata attcaatttt aacgtcttcg tccatttggg
1201 cttcaaattc atcgagtagt gctcgtgctt ctgcaattga ttgtgtgttc atcaattgat
1261 gtcgaagttc gctagcgcct cttatgccac gcacatagat tttaaagaat ctacgcaagc
1321 tcttgaattg tcgtatttca tctttttcat atttgttaaa caatgataaa tgcaatctca
1381 atagatctaa tagttccttg cttgtgtgtt cgcgtggttc tttttcaaaa gcgaatggat
1441 tgtggaaaat gcctctacca atcatgacgc catcaatgcc atatttttct gccagttcaa
1501 gtcctgtttt tctatcggga atatcaccgt taattgttaa caatgtattt ggtgcaattt
1561 cgtcacgtaa atttttaata gcttcgatta attcccaatg tgcatctact ttactcattt
1621 ctttacgtgt acgaagatga atagataaat tggcaatgtc ttgttcgaag acgtgcttca
1681 accaatcttt ccattcatcg atttcatagt agccaaggcg tgttttaaca cttaccggaa
1741 gcccacctgc tttagtcgct tgaataattt cggcagcaac gtcaggtctt aagattaagc
1801 cggaaccctt acggtacccg gggatcctct agagtcgacc tgcaggcatg caagctctga
1861 tttcactttt tgcattctac aaactgcata actcatatgt aaatcgctcc tttttaggtg
1921 gcacaaatgt gaggcatttt cgctctttcc ggcgaggcta gttaccctta agttattggt
1981 atgactggtt ttaagcgcaa aaaaagttgc tttttcgtac ctattaatgt atcgttagaa
2041 aaccgactgt aaaaagtaca gtcggcatta tctcatatta taaaagccag tcattaggcc
2101 tatctgacaa ttcctgaata gagttcataa acaatcctgc atgataacca tcacaaacag
2161 aatgatgtac ctgtaaagat agcggtaaat atattgaatt acctttatta atgaattttc
2221 ctgctgtaat aatgggtaga aggtaattac tattattatt gatatttaag ttaaacccag
2281 taaatgaagt ccatggaata atagaaagag aaaaagcatt ttcaggtata ggtgttttgg
2341 gaaacaattt ccccgaacca ttatatttct ctacatcaga aaggtataaa tcataaaact
2401 ctttgaagtc attctttaca ggagtccaaa taccagagaa tgttttagat acaccatcaa
2461 aaattgtata aagtggctct aacttatccc aataacctaa ctctccgtcg ctattgtaac
2521 cagttctaaa agctgtattt gagtttatca cccttgtcac taagaaaata aatgcagggt
2581 aaaatttata tccttcttgt tttatgtttc ggtataaaac actaatatca atttctgtgg
2641 ttatactaaa agtcgtttgt tggttcaaat aatgattaaa tatctctttt ctcttccaat
2701 tgtctaaatc aattttatta aagttcattt gatatgcctc ctaaattttt atctaaagtg
2761 aatttaggag gcttacttgt ctgctttctt cattagaatc aatccttttt taaaagtcaa
2821 tattactgta acataaatat atattttaaa aatatcccac tttatccaat tttcgtttgt
2881 tgaactaatg ggtgctttag ttgaagaata aagaccacat taaaaaatgt ggtcttttgt
2941 gtttttttaa aggatttgag cgtagcgaaa aatccttttc tttcttatct tgataataag
3001 ggtaactatt gccgggatag actgtaacat tctcacgcat aaaatcccct ttcattttct
3061 aatgtaaatc tattacctta ttattaattc aattcgctca taattaatcc tttttcttat
3121 tacgcaaaat ggcccgattt aagcacaccc tttattccgt taatgcgcca tgacagccat
3181 gataattact aatactagga gaagttaata aatacgggtg atctaatgat tcaatgattc
3241 aaacccttgt gaacttcttt agaacaaaag aggttcgtaa caagattttc ttcacactag
3301 caatgttagt aatttttaaa atagggactt atataccagc tccaggagta aatcctgcag
3361 cttttgataa tccccaaggt tctcaaggtg ccactgagtt attaaatact tttggtggcg
3421 gagccttgaa acgattttct atttttgcaa tgggtattgt accctacatc actgcatcaa
3481 tcgtaatgca attattacaa atggatattg tccctaaatt ctcagaatgg gcaaaacaag
3541 gtgaagtagg tagaagaaag ttaaataacg ttactcgtta tttagcaatt tctttagcat
3601 ttatccaatc tataggtatg gcattccaat ttaataatta tctcaaaggt gcgctgatta
3661 tcaatcagtc aattatgagt tatttattaa tagcactagt tttgacagca ggaactgctt
3721 tcttaatatg gcttggtgat caaatcactc agttcggtgt tggtaatggt atttctatta
3781 tcatattccc atcaagctta ttttaattat actctatcaa tgatagagtg tcaatatttt
3841 ttttagtttt tcatgaactc gatcgagggg atccaaataa aaaactagtt tgacaaataa
3901 ctctatcaat gatagagtgt caacaaaaag gaggaattaa tgatgtctag attagataaa
3961 agtaaagtga ttaacagcgc attagagctg cttaatgagg tcggaatcga aggtttaaca
4021 acccgtaaac tcgcccagaa gctaggtgta gagcagccta cattgtattg gcatgtaaaa
4081 aataagcggg ctttgctcga cgccttagcc attgagatgt tagataggca ccatactcac
4141 ttttgccctt tagaagggga aagctggcaa gattttttac gtaataacgc taaaagtttt
4201 agatgtgctt tactaagtca tcgcgatgga gcaaaagtac atttaggtac acggcctaca
4261 gaaaaacagt atgaaactct cgaaaatcaa ttagcctttt tatgccaaca aggtttttca
4321 ctagagaatg cattatatgc actcagcgct gtggggcatt ttactttagg ttgcgtattg
4381 gaagatcaag agcatcaagt cgctaaagaa gaaagggaaa cacctactac tgatagtatg
4441 ccgccattat tacgacaagc tatcgaatta tttgatcacc aaggtgcaga gccagccttc
4501 ttattcggcc ttgaattgat catatgcgga ttagaaaaac aacttaaatg tgaaagtggg
4561 tcttaaaagc agcataacct ttttccgtga tggtaacttc acggtaacca agataacaaa
4621 gaatacaaga aaatatttac aaaaaatcaa tttaacaatt ccttaaaaca tgcaggaatt
4681 gacgatttaa acaatattag ctttgaacaa ttcttatctc ttttcaatag ctataaatta
4741 tttaataagt aagttaaggg atgcataaac tgcatccctt aacttgtttt tcgtgtgcct
4801 attttttgtg aatcgattat gtcttttgcg cagtcggctt aaaccagttt tcgctggtgc
4861 gaaaaaagag tgtcttgtga cacctaaatt caaaatctat cggtcagatt tataccgatt
4921 tgattttata tattcttgaa taacatacgc cgagttatca cataaaagcg ggaaccaatc
4981 atcaaattta aacttcattg cataatccat taaactctta aattctacga ttccttgttc
5041 atcaataaac tcaatcattt ctttaattaa tttatatcta tctgttgttg ttttctttaa
5101 taattcatca acatctacac cgccataaac tatcatatct tctttttgat atttaaattt
5161 attaggatcg tccatgtgaa gcatatatct cacaagacct ttcacacttc ctgcaatctg
5221 cggaatagtc gcattcaatt cttctgtaat tatttttatc tgttcataag atttattacc
5281 ctcatacatc actagaatat gataatgctc ttttttcatc ctatcttctg tatcagtatc
5341 cctatcatgt aatggagaca ctacaaattg aatgtgtaac tcttttaaat actctaacca
5401 ctcggctttt gctgattctg gatataaaac aaatgtccaa ttacgtcctc ttgaattttt
5461 cttgttttca gtttctttta ttacattttc gctcatgata taataacggt gctaatacat
5521 ttaacaaaat ttagtcatag ataggcagca tgccagtgct gtctatcttt ttttgtttaa
5581 aatgcaccgt attcctcctt tgcatatttt tttattagaa taccggttgc atctgatttg
5641 ctaatattat atttttcttt gattctattt aatatctcat tttcttctgt tgtaagtctt
5701 aaagtaacag caactttttt ctcttctttt ctatctacaa ccatcactgt acctcccaac
5761 atctgttttt ttcactttaa cataaaaaac aaccttttaa cattaaaaac ccaatattta
5821 tttatttgtt tggacaatgg acaatggaca cctagggggg aggtcgtagt acccccctat
5881 gttttctccc ctaaataacc ccaaaaatct aagaaaaaaa gacctcaaaa aggtctttaa
5941 ttaacatctc aaatttcgca tttattccaa tttccttttt gcgtgtgatg cgctgcgtcc
6001 attaaaaatc ctagagcttt gcaaccgaaa gttaatagct gtcgctacta ctttcgctta
6061 cgctctaagt atattttaag gactgtcaca cgcaaaaagt tttctcggca taaaagtacc
6121 tctacatctc taaatcgtct gtacgctgtt tctcacgctt tctatcgacc ttctggacat
6181 tatcctgtac aacatccata aactgtccca cacgctcgaa tttggaatca ttaaagaatt
6241 tctctttaag cctattaaac cctttctcaa acccagggaa attcgccctc gcagcacgat
6301 ataaagtcac tgtactatct tgaaatttct ctgatacatt caactgctca ttcaaactat
6361 cattctctcg ctttaattta ttaacctctt tacttttttc gtgatacccc tctttccatg
6421 tattcactac ttctttcaaa ctctctctac gtttttttaa ttcttgattt tctgtgtaat
6481 agtctgtgct cttaatattt tcgtaatcat caacaatccg ttcctgcctc gcgcgtttcg
6541 gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt
6601 aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc
6661 ggggcgcagc catgacccag tcacgtagcg atagcggagt gtatactggc ttaactatgc
6721 ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg
6781 cgtaaggaga aaataccgca tcaggcgctc ttccgcttcc tcgctcactg actcgctgcg
6841 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
6901 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
6961 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
7021 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
7081 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
7141 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
7201 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
7261 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
7321 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
7381 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt
7441 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
7501 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
7561 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
7621 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
7681 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
7741 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
7801 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
7861 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
7921 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
7981 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
8041 tttgcgcaac gttgttgcca ttgctgcagg catcgtggtg tcacgctcgt cgtttggtat
8101 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
8161 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
8221 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
8281 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
8341 accgagttgc tcttgcccgg cgtcaacacg ggataatacc gcgccacata gcagaacttt
8401 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
8461 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
8521 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
8581 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
8641 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
8701 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat
8761 tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc ttcaa
//
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pJB38-NWMN2930质粒载体应用举例
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