| 质粒名称: | pMRE132 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 118486 |
| 存储实验室: | Mitja Remus-Emsermann |
|
载体骨架基本信息 |
|
| 载体名称: | pFru97 |
| 载体出品公司: | Clontech |
| 完整质粒大小: | 7584 bp |
| 载体类型: | 细菌表达 |
| 筛选标记: | Chloramphenicol |
| 载体抗性: | Kanamycin |
| 生长温度: | 37 ℃ |
| 宿主菌: | E. coli S17-1 lambda pir |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | sGFP2 |
| 物种来源: | Synthetic |
| 克隆方法: | Gibson Cloning |
|
文献引用方法 |
|
| 材料和方法部分: | pMRE132 was a gift from Mitja Remus-Emsermann (Addgene plasmid # 118486 ; http://n2t.net/addgene:118486 ; RRID:Addgene_118486) |
| 参考文献部分: | Chromatic Bacteria - A Broad Host-Range Plasmid and Chromosomal Insertion Toolbox for Fluorescent Protein Expression in Bacteria. Schlechter RO, Jun H, Bernach M, Oso S, Boyd E, Munoz-Lintz DA, Dobson RCJ, Remus DM, Remus-Emsermann MNP. Front Microbiol. 2018 Dec 12;9:3052. doi: 10.3389/fmicb.2018.03052. eCollection 2018. 10.3389/fmicb.2018.03052 PubMed 30631309 |
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 pMRE132 7584 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION broad-host range plasmid to constitutively express fluorescent
protein genes in bacteria.
ACCESSION .
VERSION .
KEYWORDS pMRE132
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7584)
AUTHORS Schlechter RO, Jun H, Bernach M, Oso S, Boyd E, Munoz-Lintz DA,
Dobson RCJ, Remus DM, Remus-Emsermann MNP
TITLE Chromatic Bacteria - A Broad Host-Range Plasmid and Chromosomal
Insertion Toolbox for Fluorescent Protein Expression in Bacteria.
JOURNAL Front Microbiol. 2018 Dec 12;9:3052. doi: 10.3389/fmicb.2018.03052.
eCollection 2018.
PUBMED 30631309
REFERENCE 2 (bases 1 to 7584)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..7584
/organism="synthetic DNA construct"
/mol_type="other DNA"
CDS 1..516
/codon_start=1
/product="N-terminal fragment of mVenus for use in
bimolecular fluorescence complementation (BiFC) (Kodama and
Hu, 2010)"
/label=VN173
/translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
NFKIRHNIE"
primer_bind complement(43..64)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(304..323)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 651..672
/label=EGFP-C
/note="EGFP, forward primer"
terminator 701..795
/label=lambda t0 terminator
/note="transcription terminator from phage lambda"
CDS 839..1498
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/label=CmR
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
primer_bind complement(905..924)
/label=CAT-R
/note="Chloramphenicol resistance gene, reverse primer"
terminator 1563..1649
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
CDS complement(1988..2782)
/codon_start=1
/gene="aph(3')-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/label=NeoR/KanR
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(2099..2118)
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind 2709..2728
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
rep_origin 4262..5033
/label=pBBR1 oriV
/note="replication origin of the broad-host-range plasmid
pBBR1 from Bordetella bronchiseptica; requires the pBBR1
Rep protein for replication"
CDS 5034..5696
/codon_start=1
/product="replication protein for the broad-host-range
plasmid pBBR1 from Bordetella bronchiseptica"
/label=pBBR1 Rep
/translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
primer_bind 6390..6412
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 6404..6421
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 6405..6421
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 6431..6450
/label=T7
/note="T7 promoter, forward primer"
promoter 6431..6449
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
terminator 6492..6578
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 6675..6761
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 6858..6944
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 7041..7127
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
ORIGIN
1 atgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc
61 gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc
121 aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc
181 gtgaccaccc tgacctacgg cgtgcagtgc ttcgcccgct accccgacca catgaagcag
241 cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc
301 aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg
361 aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag
421 ctggagtaca actacaacag ccacaacgtc tatatcaccg ccgacaagca gaagaacggc
481 atcaaggcca acttcaagat ccgccacaac atcgaggacg gcggcgtgca gctcgccgac
541 cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac
601 ctgagcaccc agtccaagct gagcaaagac cccaacgaga agcgcgatca catggtcctg
661 ctggagttcg tgaccgccgc cgggtaatta gctgagcttg gactcctgtt gatagatcca
721 gtaatgacct cagaactcca tctggatttg ttcagaacgc tcggttgccg ccgggcgttt
781 tttattggtg agaatccaag ctagcttggc gagattttca ggagctaagg aagctaaaat
841 ggagaaaaaa atcactggat ataccaccgt tgatatatcc caatggcatc gtaaagaaca
901 ttttgaggca tttcagtcag ttgctcaatg tacctataac cagaccgttc agctggatat
961 tacggccttt ttaaagaccg taaagaaaaa taagcacaag ttttatccgg cctttattca
1021 cattcttgcc cgcctgatga atgctcatcc ggaatttcgt atggcaatga aagacggtga
1081 gctggtgata tgggatagtg ttcacccttg ttacaccgtt ttccatgagc aaactgaaac
1141 gttttcatcg ctctggagtg aataccacga cgatttccgg cagtttctac acatatattc
1201 gcaagatgtg gcgtgttacg gtgaaaacct ggcctatttc cctaaagggt ttattgagaa
1261 tatgtttttc gtctcagcca atccctgggt gagtttcacc agttttgatt taaacgtggc
1321 caatatggac aacttcttcg cccccgtttt caccatgggc aaatattata cgcaaggcga
1381 caaggtgctg atgccgctgg cgattcaggt tcatcatgcc gtttgtgatg gcttccatgt
1441 cggcagaatg cttaatgaat tacaacagta ctgcgatgag tggcagggcg gggcgtaatt
1501 tttttaaggc agttattggt gcccttaaac gcctggggta atgactctct agcttgaggc
1561 atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc tgttgtttgt
1621 cggtgaacgc tctcctgagt aggacaaatc cgccgctagg agcttgcggc cgccagtgtg
1681 atggatatct gcataaaaac tgttgtaatt cattaagcat tctgccgaca tggaagccat
1741 cacaaacggc atgatgaacc tgaatcgcca gcggcatcag caccttgtcg ccttgcgtat
1801 aatatttgcc catgggggtg ggcgaagaac tccagcatga gatccccgcg ctggaggatc
1861 atccagccgg cgtcccggaa aacgattccg aagcccaacc tttcatagaa ggcggcggtg
1921 gaatcgaaat ctcgtgatgg caggttgggc gtcgcttggt cggtcatttc gaaccccaga
1981 gtcccgctca gaagaactcg tcaagaaggc gatagaaggc gatgcgctgc gaatcgggag
2041 cggcgatacc gtaaagcacg aggaagcggt cagcccattc gccgccaagc tcttcagcaa
2101 tatcacgggt agccaacgct atgtcctgat agcggtccgc cacacccagc cggccacagt
2161 cgatgaatcc agaaaagcgg ccattttcca ccatgatatt cggcaagcag gcatcgccat
2221 gggtcacgac gagatcctcg ccgtcgggca tgcgcgcctt gagcctggcg aacagttcgg
2281 ctggcgcgag cccctgatgc tcttcgtcca gatcatcctg atcgacaaga ccggcttcca
2341 tccgagtacg tgctcgctcg atgcgatgtt tcgcttggtg gtcgaatggg caggtagccg
2401 gatcaagcgt atgcagccgc cgcattgcat cagccatgat ggatactttc tcggcaggag
2461 caaggtgaga tgacaggaga tcctgccccg gcacttcgcc caatagcagc cagtcccttc
2521 ccgcttcagt gacaacgtcg agcacagctg cgcaaggaac gcccgtcgtg gccagccacg
2581 atagccgcgc tgcctcgtcc tgcagttcat tcagggcacc ggacaggtcg gtcttgacaa
2641 aaagaaccgg gcgcccctgc gctgacagcc ggaacacggc ggcatcagag cagccgattg
2701 tctgttgtgc ccagtcatag ccgaatagcc tctccaccca agcggccgga gaacctgcgt
2761 gcaatccatc ttgttcaatc atgcgaaacg atcctcatcc tgtctcttga tcagatcttg
2821 atcccctgcg ccatcagatc cttggcggca agaaagccat ccagtttact ttgcagggct
2881 tcccaacctt accagagggc gccccagctg gcaattccgg ttcgcttgct gtccataaaa
2941 ccgcccagtc tagctatcgc catgtaagcc cactgcaagc tacctgcttt ctctttgcgc
3001 ttgcgttttc ccttgtccag atagcccagt agctgacatt catcccaggt ggcacttttc
3061 ggggaaatgt gcgcgcccgc gttcctgctg gcgctgggcc tgtttctggc gctggacttc
3121 ccgctgttcc gtcagcagct tttcgcccac ggccttgatg atcgcggcgg ccttggcctg
3181 catatcccga ttcaacggcc ccagggcgtc cagaacgggc ttcaggcgct cccgaaggtc
3241 tcgggccgtc tcttgggctt gatcggcctt cttgcgcatc tcacgcgctc ctgcggcggc
3301 ctgtagggca ggctcatacc cctgccgaac cgcttttgtc agccggtcgg ccacggcttc
3361 cggcgtctca acgcgctttg agattcccag cttttcggcc aatccctgcg gtgcataggc
3421 gcgtggctcg accgcttgcg ggctgatggt gacgtggccc actggtggcc gctccagggc
3481 ctcgtagaac gcctgaatgc gcgtgtgacg tgccttgctg ccctcgatgc cccgttgcag
3541 ccctagatcg gccacagcgg ccgcaaacgt ggtctggtcg cgggtcatct gcgctttgtt
3601 gccgatgaac tccttggccg acagcctgcc gtcctgcgtc agcggcacca cgaacgcggt
3661 catgtgcggg ctggtttcgt cacggtggat gctggccgtc acgatgcgat ccgccccgta
3721 cttgtccgcc agccacttgt gcgccttctc gaagaacgcc gcctgctgtt cttggctggc
3781 cgacttccac cattccgggc tggccgtcat gacgtactcg accgccaaca cagcgtcctt
3841 gcgccgcttc tctggcagca actcgcgcag tcggcccatc gcttcatcgg tgctgctggc
3901 cgcccagtgc tcgttctctg gcgtcctgct ggcgtcagcg ttgggcgtct cgcgctcgcg
3961 gtaggcgtgc ttgagactgg ccgccacgtt gcccattttc gccagcttct tgcatcgcat
4021 gatcgcgtat gccgccatgc ctgcccctcc cttttggtgt ccaaccggct cgacgggggc
4081 agcgcaaggc ggtgcctccg gcgggccact caatgcttga gtatactcac tagactttgc
4141 ttcgcaaagt cgtgaccgcc tacggcggct gcggcgccct acgggcttgc tctccgggct
4201 tcgccctgcg cggtcgctgc gctcccttgc cagcccgtgg atatgtggac gatggccgcg
4261 agcggccacc ggctggctcg cttcgctcgg cccgtggaca accctgctgg acaagctgat
4321 ggacaggctg cgcctgccca cgagcttgac cacagggatt gcccaccggc tacccagcct
4381 tcgaccacat acccaccggc tccaactgcg cggcctgcgg ccttgcccca tcaatttttt
4441 taattttctc tggggaaaag cctccggcct gcggcctgcg cgcttcgctt gccggttgga
4501 caccaagtgg aaggcgggtc aaggctcgcg cagcgaccgc gcagcggctt ggccttgacg
4561 cgcctggaac gacccaagcc tatgcgagtg ggggcagtcg aagggcgaag cccgcccgcc
4621 tgccccccga gcctcacggc ggcgagtgcg ggggttccaa gggggcagcg ccaccttggg
4681 caaggccgaa ggccgcgcag tcgatcaaca agccccggag gggccacttt ttgccggagg
4741 gggagccgcg ccgaaggcgt gggggaaccc cgcaggggtg cccttctttg ggcaccaaag
4801 aactagatat agggcgaaat gcgaaagact taaaaatcaa caacttaaaa aaggggggta
4861 cgcaacagct cattgcggca ccccccgcaa tagctcattg cgtaggttaa agaaaatctg
4921 taattgactg ccacttttac gcaacgcata attgttgtcg cgctgccgaa aagttgcagc
4981 tgattgcgca tggtgccgca accgtgcggc acccctaccg catggagata agcatggcca
5041 cgcagtccag agaaatcggc attcaagcca agaacaagcc cggtcactgg gtgcaaacgg
5101 aacgcaaagc gcatgaggcg tgggccgggc ttattgcgag gaaacccacg gcggcaatgc
5161 tgctgcatca cctcgtggcg cagatgggcc accagaacgc cgtggtggtc agccagaaga
5221 cactttccaa gctcatcgga cgttctttgc ggacggtcca atacgcagtc aaggacttgg
5281 tggccgagcg ctggatctcc gtcgtgaagc tcaacggccc cggcaccgtg tcggcctacg
5341 tggtcaatga ccgcgtggcg tggggccagc cccgcgacca gttgcgcctg tcggtgttca
5401 gtgccgccgt ggtggttgat cacgacgacc aggacgaatc gctgttgggg catggcgacc
5461 tgcgccgcat cccgaccctg tatccgggcg agcagcaact accgaccggc cccggcgagg
5521 agccgcccag ccagcccggc attccgggca tggaaccaga cctgccagcc ttgaccgaaa
5581 cggaggaatg ggaacggcgc gggcagcagc gcctgccgat gcccgatgag ccgtgttttc
5641 tggacgatgg cgagccgttg gagccgccga cacgggtcac gctgccgcgc cggtagcact
5701 tgggttgcgc agcaacccgt aagtgcgctg ttccagacta tcggctgtag ccgcctcgcc
5761 gccctatacc ttgtctgcct ccccgcgttg cgtcgcggtg catggagccg ggccacctcg
5821 acctgaatgg aagccggcgg cacctcgcta acggattcac cgtttttatc aggctctggg
5881 aggcagaata aatgatcata tcgtcaatta ttacctccac ggggagagcc tgagcaaact
5941 ggcctcaggc atttgagaag cacacggtca cactgcttcc ggtagtcaat aaaccggtaa
6001 accagcaata gacataagcg gctatttaac gaccctgccc tgaaccgacg accgggtcga
6061 atttgctttc gaatttctgc cattcatccg cttattatca cttattcagg cgtagcaacc
6121 aggcgtttaa gggcaccaat aactgcctta aaaaaattac gccccgccct gccactcatc
6181 gcagtacggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
6241 acaaaatatt aacgcttaca atttccattc gccattcagg ctgcgcaact gttgggaagg
6301 gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag
6361 gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag
6421 tgagcgcgcg taatacgact cactataggg cgaattggag ctggggccgc aattcccaat
6481 tccccaggca tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt cgttttatct
6541 gttgtttgtc ggtgaacgct ctcctgagta ggacaaatcc gccgggagcg gatttgaacg
6601 ttgcgaagca acggcccgga gggtggcggg caggacgccc gccataaact gccaggaatt
6661 aattccccag gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta
6721 tctgttgttt gtcggtgaac gctctcctga gtaggacaaa tccgccggga gcggatttga
6781 acgttgcgaa gcaacggccc ggagggtggc gggcaggacg cccgccataa actgccagga
6841 attaattccc caggcatcaa ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt
6901 ttatctgttg tttgtcggtg aacgctctcc tgagtaggac aaatccgccg ggagcggatt
6961 tgaacgttgc gaagcaacgg cccggagggt ggcgggcagg acgcccgcca taaactgcca
7021 ggaattaatt ccccaggcat caaataaaac gaaaggctca gtcgaaagac tgggcctttc
7081 gttttatctg ttgtttgtcg gtgaacgctc tcctgagtag gacaaatccg ccgggagcgg
7141 atttgaacgt tgcgaagcaa cggcccggag ggtggcgggc aggacgcccg ccataaactg
7201 ccaggaattg gggatcggat tcgagctcgc acgctgccgc aagcactcag ggcgcaaggg
7261 ctgctaaagg aagcggaaca cgtagaaagc cagtccgcag aaacggtgct gaccccggat
7321 gaatgtcagc tactgggcta tctggacaag ggaaaacgca agcgcaaaga gaaagcaggt
7381 agcttgcagt gggcttacat ggcgatagct agactgggcg gttttatgga cagcaagcga
7441 accggaattg ccagctgggg cgccctctgg taaggttggg aagccctgca aagtaaactg
7501 gatggctttc ttgccgccaa ggatctgatg gcgcagggga tcaagatctg atcaagagac
7561 aggatactag tggaggaaga aaaa
//
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