pCT5-bac2.0

价格:1200元

联系方式:I47-825O-882O

相关技术服务:质粒构建    基因合成    质粒大提

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pCT5bac2.0载体基本信息

质粒名称: pCT5-bac2.0
出品公司: Addgene
目录编号: 119872
存储实验室: Claudia Schmidt-dannert

载体骨架基本信息
载体名称: pRBBm34
载体出品公司: Biedendieck et al., 2007
空载体大小: --
完整质粒大小: --
载体修饰: --
载体类型: 细菌表达,合成生物学
筛选标记: --
载体抗性: 氨苄霉素
生长温度: 37 ℃
宿主菌: DH5a
拷贝数: --

插入基因信息
插入基因名称: CymR
物种来源: Pseudomonas putida F1
插入大小: 624 bp
基因突变: --
GenBank编号 --
启动子: PxylR promoter
标签/融合蛋白: --
克隆方法: Gibson克隆
5’克隆位点: --
3’克隆位点: --
5’测序引物 tagagagagaatataaaaagccaTTACTAGCGCTTGAATTTC
3’测序引物 --

文献引用方法
材料和方法部分: pCT5-bac2.0 was a gift from Claudia Schmidt-dannert (Addgene plasmid # 119872 ; http://n2t.net/addgene:119872 ; RRID:Addgene_119872)
参考文献部分: Development of a synthetic cumate-inducible gene expression system for Bacillus. Seo SO, Schmidt-Dannert C. Appl Microbiol Biotechnol. 2018 Nov 3. pii: 10.1007/s00253-018-9485-4. doi: 10.1007/s00253-018-9485-4. 10.1007/s00253-018-9485-4 PubMed 30392122

序号

pCT5bac2.0载体图谱质粒图谱和多克隆位点信息

pCT5-bac2.0载体图谱

pCT5bac2.0载体简介

pCT5bac2.0载体序列

LOCUS       pCT5-bac2.0                7877 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Cumate-inducible expression of sfGFP gene in Bacillus subtilis, B. 
            megaterium, and Escherichia coli.
ACCESSION   .
VERSION     .
KEYWORDS    pCT5-bac2.0
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7877)
  AUTHORS   Seo SO, Schmidt-Dannert C
  TITLE     Development of a synthetic cumate-inducible gene expression system 
            for Bacillus.
  JOURNAL   Appl Microbiol Biotechnol. 2018 Nov 3. pii: 
            10.1007/s00253-018-9485-4. doi: 10.1007/s00253-018-9485-4.
  PUBMED    30392122
REFERENCE   2  (bases 1 to 7877)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7877
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     terminator      14..41
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(77..688)
                     /codon_start=1
                     /gene="cymR from Pseudomonas putida"
                     /product="cumate repressor (Mullick et al., 2006)"
                     /label=CymR
                     /translation="MSPKRRTQAERAMETQGKLIAAALGVLREKGYAGFRIADVPGAAG
                     VSRGAQSHHFPTKLELLLATFEWLYEQITERSRARLAKLKPEDDVIQQMLDDAAEFFLD
                     DDFSISLDLIVAADRDPALREGIQRTVERNRFVVEDMWLGVLVSRGLSRDDAEDILWLI
                     FNSVRGLAVRSLWQKDKERFERVRNSTLEIARERYAKFKR"
     protein_bind    1026..1053
                     /label=CuO
                     /bound_moiety="CymR"
                     /note="CymR-binding P2 operator sequence from the p-cmt 
                     operon of Pseudomonas putida (Mullick et al., 2006)"
     CDS             1082..1798
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /note="mammalian codon-optimized"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     terminator      1849..1920
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      complement(2101..2689)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2181..2200)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2860..3720)
                     /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     3483..3502
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3721..3825)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3893..3911
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             complement(3989..5365)
                     /codon_start=1
                     /gene="Bacillus subtilis tetL"
                     /product="tetracycline efflux protein"
                     /label=TcR
                     /note="confers resistance to tetracycline"
                     /translation="MNTSYSQSNLRHNQILIWLCILSFFSVLNEMVLNVSLPDIANDFN
                     KPPASTNWVNTAFMLTFSIGTAVYGKLSDQLGIKRLLLFGIIINCFGSVIGFVGHSFFS
                     LLIMARFIQGAGAAAFPALVMVVVARYIPKENRGKAFGLIGSIVAMGEGVGPAIGGMIA
                     HYIHWSYLLLIPMITIITVPFLMKLLKKEVRIKGHFDIKGIILMSVGIVFFMLFTTSYS
                     ISFLIVSVLSFLIFVKHIRKVTDPFVDPGLGKNIPFMIGVLCGGIIFGTVAGFVSMVPY
                     MMKDVHQLSTAEIGSVIIFPGTMSVIIFGYIGGILVDRRGPLYVLNIGVTFLSVSFLTA
                     SFLLETTSWFMTIIIVFVLGGLSFTKTVISTIVSSSLKQQEAGAGMSLLNFTSFLSEGT
                     GIAIVGGLLSIPLLDQRLLPMEVDQSTYLYSNLLLLFSGIIVISWLVTLNVYKHSQRDF
                     "
     CDS             complement(5565..6569)
                     /codon_start=1
                     /gene="repB"
                     /product="RepB replication protein"
                     /label=repB
                     /note="from Enterococcus faecalis plasmid pAM-alpha-1"
                     /db_xref="GI:22652809"
                     /protein_id="AAN03827.1"
                     /translation="MGVSFNIMCPNSSIYSDEKSRVLVDKTKSGKVRPWREKKIANVDY
                     FELLHILEFKKAERVKDCAEILEYKQNRETGERKLYRVWFCKSRLCPMCNWRRAMKHGI
                     QSQKVVAEVIKQKPTVRWLFLTLTVKNVYDGEELNKSLSDMAQGFRRMMQYKKINKNLV
                     GFMRATEVTINNKDNSYNQHMHVLVCVEPTYFKNTENYVNQKQWIQFWKKAMKLDYDPN
                     VKVQMIRPKNKYKSDIQSAIDETAKYPVKDTDFMTDDEEKNLKRLSDLEEGLHRKRLIS
                     YGGLLKEIHKKLNLDDTEEGDLIHTDDDEKADEDGFSIIAMWNWERKNYFIKE"
     promoter        6994..7022
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
ORIGIN
        1 actagttcac actggctcac cttcgggtgg gcctttctgc gtttatatac tagagagaga
       61 atataaaaag ccattactag cgcttgaatt tcgcgtaccg ctctcgcgca atttcgagtg
      121 tcgagttcct gacacgctca aagcgttctt tgtccttctg ccataggcta cgaacagcaa
      181 gcccacgcac cgaattgaat atcaaccaaa ggatatcttc tgcatcatca cgcgaaagac
      241 cacggctcac cagaacacca agccacatat cctcgacgac aaaccgattc ctctctaccg
      301 tgcgctgaat accctcgcgt aacgctgggt cccggtcggc agccacaatc aaatcaaggc
      361 tgatagagaa gtcatcgtcg aggaaaaatt cggcggcgtc gtccagcatt tgctggatga
      421 cgtcatcctc tggcttcaat ttcgctaatc gagcccgact gcgttcggtg atctgttcgt
      481 aaagccattc aaaagtggca agcagaagct caagctttgt cgggaaatga tggctctgcg
      541 ctcctctgga gacaccagca gcaccgggca catctgcgat ccggaatccc gcgtaacctt
      601 tttcccgtaa aacccccagg gccgctgcaa tcaacttgcc ctgggtctcc attgcgcgct
      661 ctgcctgtgt tcttctcttt ggactcatga tcaccatttt ttttactaaa gcttgatctg
      721 caatttgaat aataaccact cctttgttta tccaccgaac taagttggtg ttttttgaag
      781 cttgaattag atatttaaaa gtatcatatc taatattata actaaatttt ctaaaaaaaa
      841 cattgaagga acggttattt gattattaat ccggcttttt tattatttag gcaactgaaa
      901 cgattcggat cctgtattac tattctattt tgtcaaaata attttattga caacgtctta
      961 ttaacgttga tataatttaa attttatttg acaaaaatgg gctcgtgttg tacaataaat
     1021 gtagtaacaa acagacaatc tggtctgttt gtattatgat taactaataa ggaggacaaa
     1081 catgtcaaaa ggagaagaac tttttacagg tgtagtacct atcttggttg aattggatgg
     1141 tgatgttaac ggtcacaaat tttctgtacg tggtgaaggt gaaggtgatg caactaacgg
     1201 taaattgaca cttaaattca tttgtacaac tggaaaactt cctgttcctt ggcctactct
     1261 tgttacaaca ttgacatatg gagtacaatg tttttcacgt tatcctgatc atatgaaacg
     1321 tcacgatttt tttaaatctg ctatgccaga aggttatgta caagaacgta caatttcatt
     1381 taaagatgac ggaacatata aaacacgtgc tgaagtaaaa ttcgaaggtg acactcttgt
     1441 taatcgtatc gaattgaaag gaatcgattt caaagaagat ggtaacattt tgggacacaa
     1501 acttgaatac aacttcaact ctcataatgt ttatatcaca gctgacaaac aaaaaaacgg
     1561 tattaaagct aattttaaaa ttcgtcacaa tgttgaagat ggatctgttc aattggctga
     1621 tcattatcaa caaaatacac caatcggaga cggaccagta ttgcttccag ataaccacta
     1681 cctttctact caatcagttc tttcaaaaga tcctaacgaa aaacgtgacc atatggtact
     1741 tcttgaattt gttacagcag caggtatcac tcacggtatg gacgaacttt ataaataagt
     1801 taaaagtctc gtaaagcgtt ctatcaataa cccgttggtg ccaggcatca aataaaacga
     1861 aaggctcagt cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc
     1921 tactagagga gctcatgccg gcggcacctc gctaacggat tcaccactcc aagaattgga
     1981 gccaatcaat tcttgcggag aactgtgaat gcgcaaacca acccttggca gaacatatcc
     2041 atcgcgtccg ccatctccag cagccgcacg cggcgcatct cgggccgcgt tgctggcgtt
     2101 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     2161 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     2221 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     2281 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     2341 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     2401 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     2461 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     2521 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac
     2581 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     2641 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     2701 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     2761 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     2821 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     2881 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     2941 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     3001 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     3061 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     3121 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctgcagg
     3181 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     3241 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     3301 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     3361 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     3421 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaacacg
     3481 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     3541 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     3601 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     3661 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     3721 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     3781 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     3841 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg
     3901 tatcacgagg ccctttcgtc ttcaagaatt cctgttataa aaaaaggatc aattttgaac
     3961 tctctcccaa agttgatccc ttaacgattt agaaatccct ttgagaatgt ttatatacat
     4021 tcaaggtaac cagccaacta atgacaatga ttcctgaaaa aagtaataac aaattactat
     4081 acagataagt tgactgatca acttccatag gtaacaacct ttgatcaagt aagggtatgg
     4141 ataataaacc acctacaatt gcaatacctg ttccctctga taaaaagctg gtaaagttaa
     4201 gcaaactcat tccagcacca gcttcctgct gtttcaagct acttgaaaca attgttgata
     4261 taactgtttt ggtgaacgaa agcccaccta aaacaaatac gattataatt gtcatgaacc
     4321 atgatgttgt ttctaaaaga aaggaagcag ttaaaaagct aacagaaaga aatgtaactc
     4381 cgatgtttaa cacgtataaa ggacctcttc tatcaacaag tatcccacca atgtagccga
     4441 aaataatgac actcattgtt ccagggaaaa taattacact tccgatttcg gcagtactta
     4501 gctggtgaac atctttcatc atataaggaa ccatagagac aaaccctgct actgttccaa
     4561 atataattcc cccacaaaga actccaatca taaaaggtat atttttccct aatccgggat
     4621 caacaaaagg atctgttact ttcctgatat gttttacaaa tatcaggaat gacagcacgc
     4681 taacgataag aaaagaaatg ctatatgatg ttgtaaacaa cataaaaaat acaatgccta
     4741 cagacattag tataattcct ttgatatcaa aatgaccttt tatccttact tctttcttta
     4801 ataatttcat aagaaacgga acagtgataa ttgttatcat aggaatgagt agaagatagg
     4861 accaatgaat ataatgggct atcattccac caatcgctgg accgactcct tctcccatgg
     4921 ctactatcga tccaataaga ccaaatgctt tacccctatt ttcctttgga atatagcgcg
     4981 caactacaac cattacgagt gctggaaatg cagctgcacc agccccttga ataaaacgag
     5041 ccataataag taaggaaaag aaagaatggc caacaaaccc aattaccgac ccgaaacaat
     5101 ttattataat tccaaatagg agtaaccttt tgatgcctaa ttgatcagat agctttccat
     5161 atacagctgt tccaatggaa aaggttaaca taaaggctgt gttcacccag tttgtactcg
     5221 caggtggttt attaaaatca tttgcaatat caggtaatga gacgttcaaa accatttcat
     5281 ttaatacgct aaaaaaagat aaaatgcaaa gccaaattaa aatttggttg tgtcgtaaat
     5341 tcgattgtga ataggatgta ttcacatttc accctccaat aatgagggca gacgtagttt
     5401 atagggttaa tgatacgctt ccctctttta attgaaccct gttacattca ttacacttca
     5461 taattaattc ctcctaaact tgattaaaac attttaccac atataaacta agttttaaat
     5521 tcagtatttc atcacttata caacaatatg gcccgtttgt tgaactactc tttaataaaa
     5581 taatttttcc gttcccaatt ccacattgca ataatagaaa atccatcttc atcggctttt
     5641 tcgtcatcat ctgtatgaat caaatcgcct tcttctgtgt catcaaggtt taatttttta
     5701 tgtatttctt ttaacaaacc accataggag attaaccttt tacggtgtaa accttcctcc
     5761 aaatcagaca aacgtttcaa attcttttct tcatcatcgg tcataaaatc cgtatccttt
     5821 acaggatatt ttgcagtttc gtcaattgcc gattgtatat ccgatttata tttatttttc
     5881 ggtcgaatca tttgaacttt tacatttgga tcatagtcta atttcattgc ctttttccaa
     5941 aattgaatcc attgtttttg attcacgtag ttttctgtat tcttaaaata agttggttcc
     6001 acacatacca atacatgcat gtgctgatta taagaattat ctttattatt tattgtcact
     6061 tccgttgcac gcataaaacc aacaagattt ttattaattt ttttatattg catcattcgg
     6121 cgaaatcctt gagccatatc tgacaaactc ttatttaatt cttcgccatc ataaacattt
     6181 ttaactgtta atgtgagaaa caaccaacga actgttggct tttgtttaat aacttcagca
     6241 acaacctttt gtgactgaat gccatgtttc attgctctcc tccagttgca cattggacaa
     6301 agcctggatt tacaaaacca cactcgatac aactttcttt cgcctgtttc acgattttgt
     6361 ttatactcta atatttcagc acaatctttt actctttcag cctttttaaa ttcaagaata
     6421 tgcagaagtt caaagtaatc aacattagcg attttctttt ctctccatgg tctcactttt
     6481 ccactttttg tcttgtccac taaaaccctt gatttttcat ctgaataaat gctactatta
     6541 ggacacataa tattaaaaga aacccccatc tatttagtta tttgtttggt cacttataac
     6601 tttaacagat ggggtttttc tgtgcaacca attttaaggg ttttcaatac tttaaaacac
     6661 atacatacca acacttcaac gcacctttca gcaactaaaa taaaaatgac gttatttcta
     6721 tatgtatcaa gataagaaag aacaagttca aaaccatcaa aaaaagacac cttttcaggt
     6781 gcttttttta ttttataaac tcattccctg atctcgactt cgttcttttt ttacctctcg
     6841 gttatgagtt agttcaaatt cgttcttttt aggttctaaa tcgtgttttt cttggaattg
     6901 tgctgtttta tcctttacct tgtctacaaa ccccttaaaa acgtttttaa aggcttttaa
     6961 gccgtctgta cgttccttaa ggaattctca tgtttgacag cttatcatcg ataagcttta
     7021 atgcggtagt ttatcacagt taaattgcta acgcagtcag gcaccgtgta tgaaatctaa
     7081 caatgcgctc atcgtcatcc tcggcaccgt caccctggat gctgtaggca taggcttggt
     7141 tatgccggta ctgccgggcc tcttgcggga tatcgtccat tccgacagca tcgccagtca
     7201 ctatggcgtg ctgctagcgc tatatgcgtt gatgcaattt ctatgcgcac ccgttctcgg
     7261 agcactgtcc gaccgctttg gccgccgccc agtcctgctc gcttcgctac ttggagccac
     7321 tatcgactac gcgatcatgg cgaccacacc cgtcctgtgg atctgacgcg tgtaactgcg
     7381 aaggtaaaac gggtgattga aaataaacta acaaatgaag aggaacgaaa gagaagtcta
     7441 gaatttgtta cgtttatatc ggatgaatta ctgcaaaatg atacggagcc tcgcacattt
     7501 attattcaga actctctttt atcgctagag aaaatccata ctctcgcaca gaccaaagag
     7561 gtagaggaat ataaagaagt cataactact ctgacaagac atgattcata atttgaaaag
     7621 caggtaaact aacccaaaag gcaagactct ccgtagttta gaaaagagca ggttgctaat
     7681 aacatataaa cagccagttg ccgttatgat aggtgactgg ctgcatggga tgaaaaggtg
     7741 agggtggaga cagacataac actcttaata gaagagggta attctttctc ttttatagaa
     7801 aatcaattaa ttgaaagtag ctccttcatt cttaagatca aacgtgatat agtttgctaa
     7861 cctttgcgtt cacttaa
//

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