| 质粒名称: | pRN11 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 84455 |
| 存储实验室: | Reindert Nijland |
|
载体骨架基本信息 |
|
| 载体名称: | pCM29 |
| 空载体大小: | 6604 bp |
| 完整质粒大小: | 6574 bp |
| 载体修饰: | sGFP替换为插入件 |
| 载体类型: | 穿梭载体E.coli-S.aureus，从SarA-P1启动子表达sGFP |
| 筛选标记: | 也含有氯霉素抗性基因，但仅在革兰氏阳性细菌中起作用。 |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | E.coli DC10b (described in doi: 10.1128/mBio.00537-12) |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | mCherry |
| 插入大小: | 739 bp |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | xmai(未破坏) |
| 3’克隆位点: | EcoRI (未破坏) |
| 5’测序引物 | TTGCATGCCTGCAGGTCGACTCTA |
| 3’测序引物 | TTATGCTTCCGGCTCGTATGTTGTGTGG |
|
文献引用方法 |
|
| 材料和方法部分: | pRN11 was a gift from Reindert Nijland (Addgene plasmid # 84455 ; http://n2t.net/addgene:84455 ; RRID:Addgene_84455) |
| 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
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 pRN11 6574 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION shuttle plasmid for sarA P1-mCherry expression.
ACCESSION .
VERSION .
KEYWORDS pRN11
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6574)
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 6574)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6574
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(1010..1026)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(1010..1026)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(1023..1045)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 1034..1050
/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)."
promoter complement(1058..1088)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 1103..1124
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(1241..1258)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(1412..2000)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(1492..1511)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(2468..2847)
/label=ISS
/note="immunostimulatory sequence from the AmpR gene;
contains unmethylated CpG dinucleotides in the context of
5'-AACGTT-3' (Sato et al., 1996)"
primer_bind 2794..2813
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(3032..3136)
/gene="bla"
/label=AmpR promoter
primer_bind 3204..3222
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
primer_bind complement(3260..3282)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind 3383..3402
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 3596..3618
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 3610..3627
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 3611..3627
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
CDS 6302..6316
/codon_start=1
/product="enterokinase recognition and cleavage site"
/label=enterokinase site
/translation="DDDDK"
ORIGIN
1 gctagcctga tatttttgac taaaccaaat gctaacccag aaatacaatc actgtgtcta
61 atgaataatt tgttttataa acactttttt gtttacttct catttttaat tagttataat
121 taactaaata atagagcatt aaatatattt aataaaactt atttaatgca aaattatgac
181 taacatatct ataataaata aagattagat atcaatatat tatcgggcaa atgtatcgag
241 caagatgcat cggatccccg ggtaccttag gaggatgtat acatatggtg agcaagggcg
301 aggaggataa tatggctatt atcaaagaat ttatgagatt caaagttcat atggaaggtt
361 cagtcaatgg tcacgaattt gaaattgagg gagaaggtga aggcagacca tatgaaggta
421 ctcaaacggc caaacttaaa gtcactaaag gaggtcctct tccatttgca tgggatattc
481 ttagtccaca atttatgtat gggtctaaag cttacgttaa acatccagcg gacattccgg
541 attatcttaa attgtcgttt ccagaaggct tcaaatggga aagagtaatg aactttgaag
601 atggaggagt cgttacagtc actcaggata gttctcttca agatggagaa tttatctaca
661 aagtcaagtt aagaggtacc aattttccct cagatggtcc agttatgcaa aagaaaacta
721 tgggatggga agcatccagt gagcgtatgt atcctgaaga tggtgccctt aaaggcgaaa
781 ttaaacaacg cctaaagctt aaagatggtg gacattatga cgctgaagtt aaaacaactt
841 acaaagcgaa gaaaccggta caacttccag gtgcatataa tgtcaacatt aaacttgata
901 ttacaagtca taatgaagat tatactattg ttgaacaata tgagagagct gaaggaagac
961 attctacagg cggcatggac gagctgtaca agtaagaatt cgtaatcatg tcatagctgt
1021 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
1081 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
1141 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
1201 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
1261 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
1321 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
1381 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
1441 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
1501 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
1561 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
1621 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
1681 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
1741 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
1801 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
1861 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
1921 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
1981 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
2041 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
2101 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
2161 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
2221 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
2281 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
2341 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
2401 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
2461 gttgcgcaac gttgttgcca ttgctacagg catcgtgggt gtcacgctcg tcgtttggta
2521 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
2581 gcaaaaaagc ggttagctct ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
2641 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
2701 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
2761 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
2821 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
2881 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
2941 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
3001 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
3061 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
3121 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta
3181 ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt
3241 tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc
3301 tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt
3361 gtcggggctg gcttaaacta tgcggcatca gagcagattg tactgagagt gcaccatatg
3421 cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca
3481 ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag
3541 ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag
3601 tcacgacgtt gtaaaacgac ggccagtgcc aagcttctgt aggtttttag gcataaaact
3661 atatgattta cccctaaatc tttaaaatgc cccttaaaat tcaaaataaa ggcatttaaa
3721 atttaaatat ttcttgtgat aaagtttgtt aaaaaggagt ggttttatga ctgttatgtg
3781 gttatcgatt ataggtatgt ggttttgtat tggaatggca ttttttgcta tcaaggttat
3841 taaaaataaa aattagacca cgcatttatg ccgagaaaat ttattgtgcg ttgagaagaa
3901 cccttaacta aacttgcaga cgaatgtcgg catagcgtga gctattaagc cgaccattcg
3961 acaagttttg ggattgttaa gggttccgag gctcaacgtc aataaagcaa ttggaataaa
4021 gaagcgaaaa aggagaagtc ggttcagaaa aagaaggata tggatctgga gctgtaatat
4081 aaaaaccttc ttcaactaac ggggcaggtt agtgacatta gaaaaccgac tgtaaaaagt
4141 acagtcggca ttatctcata ttataaaagc cagtcattag gcctatctga caattcctga
4201 atagagttca taaacaatcc tgcatgataa ccatcacaaa cagaatgatg tacctgtaaa
4261 gatagcggta aatatattga attaccttta ttaatgaatt ttcctgctgt aataatgggt
4321 agaaggtaat tactattatt attgatattt aagttaaacc cagtaaatga agtccatgga
4381 ataatagaaa gagaaaaagc attttcaggt ataggtgttt tgggaaacaa tttccccgaa
4441 ccattatatt tctctacatc agaaaggtat aaatcataaa actctttgaa gtcattcttt
4501 acaggagtcc aaataccaga gaatgtttta gatacaccat caaaaattgt ataaagtggc
4561 tctaacttat cccaataacc taactctccg tcgctattgt aaccagttct aaaagctgta
4621 tttgagttta tcacccttgt cactaagaaa ataaatgcag ggtaaaattt atatccttct
4681 tgttttatgt ttcggtataa aacactaata tcaatttctg tggttatact aaaagtcgtt
4741 tgttggttca aataatgatt aaatatctct tttctcttcc aattgtctaa atcaatttta
4801 ttaaagttca tttgatatgc ctcctaaatt tttatctaaa gtgaatttag gaggcttact
4861 tgtctgcttt cttcattaga atcaatcctt ttttaaaagt caatattact gtaacataaa
4921 tatatatttt aaaaatatcc cactttatcc aattttcgtt tgttgaacta atgggtgctt
4981 tagttgaaga ataaaagacc acattaaaaa atgtggtctt ttgtgttttt ttaaaggatt
5041 tgagcgtagc gaaaaatcct tttctttctt atcttgataa taagggtaac tattgccggc
5101 gaggctagtt acccttaagt tattggtatg actggtttta agcgcaaaaa aagttgcttt
5161 ttcgtaccta ttaatgtatc gttttaaatg actagtaaaa aacatacata gaaaggggaa
5221 aaagcaactt tttttattgt catagtttgt gaaaactaag ttgtttttat gtgttataac
5281 atggaaaagt atactgagaa aaaacaaaga aatcaagtat ttcagaaatt tattaaacgt
5341 catattggag agaatcaaat ggatttagtt gaagattgca atacatttct gtcttttgta
5401 gctgataaaa ctttagaaaa acagaaatta tataaagcta attcttgtaa aaatcgattt
5461 tgtcctgtct gtgcttggag aaaagctaga aaagatgcat tgggtttatc tttgatgatg
5521 caatatatta agcagcaaga gaaaaaggag tttatctttt taactttgac tacacctaat
5581 gtaatgagtg atgaattaga aaatgaaata aaacgttata ataattcttt tagaaaactt
5641 ataaagagaa aaaaagtagg tagtgttata aagggatatg ttcgtaagtt agagattaca
5701 tataataaaa aaagagatga ttataatcct cattttcatg tgttaattgc agtaaataaa
5761 tcgtatttca cagataaaag atattatatt agccaacaag aatggttaga tttatggcgt
5821 gatgtaacgg gcatttcaga aataacacaa gttcaagttc aaaaaataag acaaaataat
5881 aataaagaat tatatgaaat ggctaagtat tctggtaaag atagtgatta tttaataaat
5941 caaaaagtct ttgatgcatt ttataaatca cttaaaggta aacaggtatt agtttattca
6001 ggattattta aagaggctaa aaagaaatta aaaaatgggg atttagatta cttaaaagaa
6061 attgatccaa ccgaatatat ctatcaaatt ttttatattt ggaaacaaaa agagtattta
6121 gctagtgaac tttatgactt aacagaacaa gaaaaaagag aaattaatca caaaatgata
6181 gacgaaatcg aggaagaaca ataacaaaat ataagtgcta acagctgacc tcccgataac
6241 accatgtagt tattgggagg tcagctgttg aattatgcac gagtatttta aaagttattg
6301 tgatgacgac gataaacgat tatcaaaagt ataatgttaa aatgctttat tatactaacg
6361 ttatataaac attatacttt cgttatacaa attttaaccc tgttaggaac tataaaaaat
6421 catgaaaatt ttaatttgca tgtaactggg cagtgtctta aaaaatcgac actgaatttg
6481 ctcaaatttt tgtttgtaga attagaatat atttatttgg ctcatatttg ctttttaaaa
6541 gcttgcatgc ctgcaggtcg actctagagg atcc
//
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