| 质粒名称: | pRMC2 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 68940 |
| 存储实验室: | Tim Foster |
|
载体骨架基本信息 |
|
| 载体名称: | pALC2073 |
| 载体类型: | 细菌表达；tet诱导型大肠杆菌/葡萄球菌穿梭载体 |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
|
插入基因信息 |
|
| 克隆方法: | 限制性内切酶 |
| 5’测序引物 | pRMC2 SEQF: ATTCAGGCTGCGCAAC |
| 3’测序引物 | pRMC2 SEQR: TTGTTGACATTATATCATTG |
|
文献引用方法 |
|
| 材料和方法部分: | pRMC2 was a gift from Tim Foster (Addgene plasmid # 68940 ; http://n2t.net/addgene:68940 ; RRID:Addgene_68940) |
| 参考文献部分: | An improved tetracycline-inducible expression vector for Staphylococcus aureus. Corrigan RM, Foster TJ. Plasmid. 2009 Mar;61(2):126-9. doi: 10.1016/j.plasmid.2008.10.001. Epub 2008 Nov 25. 10.1016/j.plasmid.2008.10.001 PubMed 18996145 |
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 pRMC2 6555 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION tetracycline-inducible expression vector for Staphylococcus aureus.
ACCESSION .
VERSION .
KEYWORDS pRMC2
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6555)
AUTHORS Corrigan RM, Foster TJ
TITLE An improved tetracycline-inducible expression vector for
Staphylococcus aureus.
JOURNAL Plasmid. 2009 Mar;61(2):126-9. doi: 10.1016/j.plasmid.2008.10.001.
Epub 2008 Nov 25.
PUBMED 18996145
REFERENCE 2 (bases 1 to 6555)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6555
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(18..40)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind 140..159
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 368..384
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 495..511
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 562..580
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O1 for the tetR and tetA genes"
CDS 690..1313
/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(763..782)
/label=Tet-R
/note="Tetracycline resistance gene, reverse primer"
CDS complement(1625..1639)
/codon_start=1
/product="enterokinase recognition and cleavage site"
/label=enterokinase site
/translation="DDDDK"
primer_bind complement(4323..4339)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4323..4339)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(4336..4358)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 4347..4363
/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(4371..4401)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4416..4437
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(4554..4571)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4725..5313)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4805..4824)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5484..6344)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind 6107..6126
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(6345..6449)
/gene="bla"
/label=AmpR promoter
primer_bind 6517..6535
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
ORIGIN
1 ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg
61 taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt
121 cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatgcgg
181 tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgcca ttcgccattc
241 aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg
301 gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca
361 cgacgttgta aaacgacggc cagtgaattc gagctcagat ctgttaacgg taccatcaag
421 cttattttaa ttatactcat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc
481 ccagtcacga cgttgtaaaa cgacggccag tgaattcgag ctcagatctg ttaacggtac
541 catcaagctt attttaatta tactctatca atgatagagt gtcaatattt tttttagttt
601 ttcatgaact cgaggggatc caaataaaaa actagtttga caaataactc tatcaatgat
661 ataatgtcaa caaaaaggag gaattaatga tgtctagatt agataaaagt aaagtgatta
721 acagcgcatt agagctgctt aatgaggtcg gaatcgaagg tttaacaacc cgtaaactcg
781 cccagaagct aggtgtagag cagcctacat tgtattggca tgtaaaaaat aagcgggctt
841 tgctcgacgc cttagccatt gagatgttag ataggcacca tactcacttt tgccctttag
901 aaggggaaag ctggcaagat tttttacgta ataacgctaa aagttttaga tgtgctttac
961 taagtcatcg cgatggagca aaagtacatt taggtacacg gcctacagaa aaacagtatg
1021 aaactctcga aaatcaatta gcctttttat gccaacaagg tttttcacta gagaatgcat
1081 tatatgcact cagcgctgtg gggcatttta ctttaggttg cgtattggaa gatcaagagc
1141 atcaagtcgc taaagaagaa agggaaacac ctactactga tagtatgccg ccattattac
1201 gacaagctat cgaattattt gatcaccaag gtgcagagcc agccttctta ttcggccttg
1261 aattgatcat atgcggatta gaaaaacaac ttaaatgtga aagtgggtct taaaagcagc
1321 ataacctttt tccgtgatgg taacttcacg gtaaccaaga tgtcgagtta tcgataccgt
1381 cgacctgcag gcatgcaagc ttttaaaaag caaatatgag ccaaataaat atattctaat
1441 tctacaaaca aaaatttgag caaattcagt gtcgattttt taagacactg cccagttaca
1501 tgcaaattaa aattttcatg attttttata gttcctaaca gggttaaaat ttgtataacg
1561 aaagtataat gtttatataa cgttagtata ataaagcatt ttaacattat acttttgata
1621 atcgtttatc gtcgtcatca caataacttt taaaatactc gtgcataatt caacagctga
1681 cctcccaata actacatggt gttatcggga ggtcagctgt tagcacttat attttgttat
1741 tgttcttcct cgatttcgtc tatcattttg tgattaattt ctcttttttc ttgttctgtt
1801 aagtcataaa gttcactagc taaatactct ttttgtttcc aaatataaaa aatttgatag
1861 atatattcgg ttggatcaat ttcttttaag taatctaaat ccccattttt taatttcttt
1921 ttagcctctt taaataatcc tgaataaact aatacctgtt tacctttaag tgatttataa
1981 aatgcatcaa agactttttg atttattaaa taatcactat ctttaccaga atacttagcc
2041 atttcatata attctttatt attattttgt cttatttttt gaacttgaac ttgtgttatt
2101 tctgaaatgc ccgttacatc acgccataaa tctaaccatt cttgttggct aatataatat
2161 cttttatctg tgaaatacga tttatttact gcaattaaca catgaaaatg aggattataa
2221 tcatctcttt ttttattata tgtaatctct aacttacgaa catatccctt tataacacta
2281 cctacttttt ttctctttat aagttttcta aaagaattat tataacgttt tatttcattt
2341 tctaattcat cactcattac attaggtgta gtcaaagtta aaaagataaa ctcctttttc
2401 tcttgctgct taatatattg catcatcaaa gataaaccca atgcatcttt tctagctttt
2461 ctccaagcac agacaggaca aaatcgattt ttacaagaat tagctttata taatttctgt
2521 ttttctaaag ttttatcagc tacaaaagac agaaatgtat tgcaatcttc aactaaatcc
2581 atttgattct ctccaatatg acgtttaata aatttctgaa atacttgatt tctttgtttt
2641 ttctcagtat acttttccat gttataacac ataaaaacaa cttagttttc acaaactatg
2701 acaataaaaa aagttgcttt ttcccctttc tatgtatgtt ttttactatt catttaaaac
2761 gatacattaa taggtacgaa aaagcaactt tttttgcgct taaaaccagt cataccaata
2821 acttaagggt aactagcctc gccggcaata gttaccctta ttatcaagat aagaaagaaa
2881 aggatttttc gctacgctca aatcctttaa aaaaacacaa aagaccacat tttttaatgt
2941 ggtcttttat tcttcaacta aagcacccat tagttcaaca aacgaaaatt ggataaagtg
3001 ggatattttt aaaatatata tttatgttac agtaatattg acttttaaaa aaggattgat
3061 tctaatgaag aaagcagaca agtaagcctc ctaaattcac tttagataaa aatttaggag
3121 gcatatcaaa tgaactttaa taaaattgat ttagacaatt ggaagagaaa agagatattt
3181 aatcattatt tgaaccaaca aacgactttt agtataacca cagaaattga tattagtgtt
3241 ttataccgaa acataaaaca agaaggatat aaattttacc ctgcatttat tttcttagtg
3301 acaagggtga taaactcaaa tacagctttt agaactggtt acaatagcga cggagagtta
3361 ggttattggg ataagttaga gccactttat acaatttttg atggtgtatc taaaacattc
3421 tctggtattt ggactcctgt aaagaatgac ttcaaagagt tttatgattt atacctttct
3481 gatgtagaga aatataatgg ttcggggaaa ttgtttccca aaacacctat acctgaaaat
3541 gctttttctc tttctattat tccatggact tcatttactg ggtttaactt aaatatcaat
3601 aataatagta attaccttct acccattatt acagcaggaa aattcattaa taaaggtaat
3661 tcaatatatt taccgctatc tttacaggta catcattctg tttgtgatgg ttatcatgca
3721 ggattgttta tgaactctat tcaggaattg tcagataggc ctaatgactg gcttttataa
3781 tatgagataa tgccgactgt actttttaca gtcggttttc taatgtcact aacctgcccc
3841 gttagttgaa gaaggttttt atattacagc tccagatcca tatccttctt tttctgaacc
3901 gacttctcct ttttcgcttc tttattccaa ttgctttatt gacgttgagc ctcggaaccc
3961 ttaacaatcc caaaacttgt cgaatggtcg gcttaatagc tcacgctatg ccgacattcg
4021 tctgcaagtt tagttaaggg ttcttctcaa cgcacaataa attttctcgg cataaatgcg
4081 tggtctaatt tttattttta ataaccttga tagcaaaaaa tgccattcca atacaaaacc
4141 acatacctat aatcgataac cacataacag tcataaaacc actccttttt aacaaacttt
4201 atcacaagaa atatttaaat tttaaatgcc tttattttga attttaaggg gcattttaaa
4261 gatttagggg taaatcatat agttttatgc ctaaaaacct acagaagctt ggcgtaatca
4321 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
4381 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
4441 gcgttgcgct cactgtccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
4501 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
4561 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
4621 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
4681 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
4741 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
4801 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
4861 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
4921 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
4981 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
5041 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
5101 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
5161 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
5221 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
5281 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
5341 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
5401 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
5461 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
5521 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
5581 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
5641 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
5701 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
5761 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
5821 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
5881 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
5941 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
6001 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
6061 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
6121 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
6181 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
6241 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
6301 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
6361 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
6421 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
6481 taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt
6541 cgtctcgcgc gtttc
//
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