pBE2- R Plasmid

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  • Model: PVT5016
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pBE2-R

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pBE2-R,Plasmid pBE2-R,pBE2-R vector

 

pBE2-R Informaiton

 

Promoter: P43 promoter

Replicon: ori

Plasmid classification: Bacillus subtilis plasmid; free plasmid of Bacillus subtilis; secretory plasmid of Bacillus subtilis

Plasmid size: 7014bp

Prokaryotic resistance: ampicillin Amp

Cloning strain: Escherichia coli Stbl3

Culture conditions: 37 C, aerobic, LB

Expression host: Bacillus subtilis

5'Sequencing Primers: Designing Primers Based on Sequences

3'Sequencing Primers: Designing Primers Based on Sequences

 

pBE2-R Description

The shuttle expression vector pBE2R was constructed by introducing strong promoter P43 from pWB980 plasmid and introducing signal peptide and propeptide of alkaline albumin enzyme from Bacillus alkalophilus downstream. The number of plasmid copies showed that pBE2R contained about one copy in each cell of E. coli and Bacillus subtilis. Plasmid stability test showed that the plasmid passed through 30 generations in E. coli and Bacillus subtilis without loss, and could be stable in E. coli and Bacillus subtilis.

 

pBE2-R Sequence

LOCUS       Exported                7014 bp ds-DNA     circular SYN 29-SEP-2017
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pBE2R
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7014)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 29, 2017  
FEATURES             Location/Qualifiers
     source          1..7014
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        803..907
                     /gene="bla"
                     /label=AmpR promoter
     CDS             908..1768
                     /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"
     rep_origin      1939..2527
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3930..4700)
                     /codon_start=1
                     /label=KanR/NeoR
                     /translation="MRIVNGPIIMTREERMKIVHEIKERILDKYGDDVKAIGVYGSLGR
                     QTDGPYSDIEMMCVMSTEEAEFSHEWTTGEWKVEVNFDSEEILLDYASQVESDWPLTHG
                     QFFSILPIYDSGGYLEKVYQTAKSVEAQTFHDAICALIVEELFEYAGKWRNIRVQGPTT
                     FLPSLTVQVAMAGAMLIGLHHRICYTTSASVLTEAVKQSDLPSGYDHLCQFVMSGQLSD
                     SEKLLESLENFWNGIQEWTERHGYIVDVSKRIPF"
     CDS             complement(4860..5864)
                     /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"
     misc_feature    6283..6587
                     /label=P43 promoter
     misc_feature    6620..6952
                     /label=Apr sp+pro
     CDS             6983..7000
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
ORIGIN
        1 tatagtgtca cctaaatcgt atgtgtatga tacataaggt tatgtattaa ttgtagccgc
       61 gttctaacga caatatgtac aagcctaatt gtgtagcatc tggcttactg aagcagaccc
      121 tatcatctct ctcgtaaact gccgtcagag tcggtttggt tggacgaacc ttctgagttt
      181 ctggtaacgc cgttccgcac cccggaaatg gtcagcgaac caatcagcag ggtcatcgct
      241 agccagatcc tctacgccgg acgcatcgtg gccggcatca ccggcgccac aggtgcggtt
      301 gctggcgcct atatcgccga catcaccgat ggggaagatc gggctcgcca cttcgggctc
      361 atgagcgctt gtttcggcgt gggtatggtg gcaggccccg tggccggggg actgttgggc
      421 gccatctcct tgcaccattc cttgcggcgg cggtgctcaa cggcctcaac ctactactgg
      481 gctgcttcct aatgcaggag tcgcataagg gagagcgtcg atatggtgca ctctcagtac
      541 aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc
      601 gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg
      661 gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct
      721 cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg
      781 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
      841 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
      901 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
      961 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     1021 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     1081 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     1141 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     1201 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     1261 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     1321 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     1381 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     1441 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     1501 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     1561 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     1621 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     1681 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     1741 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     1801 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     1861 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     1921 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     1981 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     2041 tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc
     2101 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     2161 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
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