pNCMO2 vector (V017507)

Basic Vector Information

pNCMO2 is a shuttle vector for Brevibacillus and E. coli. Accordingly, the expression plasmid is constructed in E. coli, after which it is introduced to Brevibacillus for protein expression. The P2 promoter, derived from a cell wall protein of the host bacterium, is used as the expression promoter for pNCMO2. Because the P2 promoter does not work in E. coli, it is useful in the cloning of the target gene. However, it is an exceptionally strong promoter in Brevibacillus resulting in efficient protein production.

Vector Name:: pNCMO2

Length:: 5225 bp

Type:: Brevibacillus Expression Vector

Source/Author:: Takara Bio

Promoter:: P2

pNCMO2 vector Map

Copy Sequence View Map

pNCMO2 vector Sequence

LOCUS       Exported                5225 bp DNA     circular SYN 22-NOV-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pNCMO2
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5225)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     Created by Takara Bio Inc. http://www.takarabio.com
            Cat. No. HB112
FEATURES             Location/Qualifiers
     source          1..5225
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        61..103
                     /label=P2 promoter
     protein_bind    104..125
                     /label=Lac operator
                     /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)."
     misc_feature    258..347
                     /label=Secretion signal peptide
     misc_feature    346..409
                     /label=MCS
                     /note="multiple cloning site"
     CDS             926..1930
                     /codon_start=1
                     /label=RepB
                     /note="RepB replication protein from Enterococcus faecalis 
                     plasmid pAM-alpha-1"
                     /translation="MGVSFNIMCPNSSIYSDEKSRVLVDKTKSGKVRPWREKKIANVDY
                     FELLHILEFKKAERVKDCAEILEYKQNRETGERKLYRVWFCKSRLCPMCNWRRAMKHGI
                     QSQKVVAEVIKQKPTVRWLFLTLTVKNVYDGEELNKSLSDMAQGFRRMMQYKKINKNLV
                     GFMRATEVTINNKDNSYNQHMHVLVCVEPTYFKNTENYVNQKQWIQFWKKAMKLDYDPN
                     VKVQMIRPKNKYKSDIQSAIDETAKYPVKDTDFMTDDEEKNLKRLSDLEEGLHRKRLIS
                     YGGLLKEIHKKLNLDDTEEGDLIHTDDDEKADEDGFSIIAMWNWERKNYFIKE"
     CDS             2099..2860
                     /codon_start=1
                     /label=NmR
                     /note="Neomycin resistance gene, selection marker in 
                     Brevibacillus"
                     /translation="VNGPIIMTREERMKIVHEIKERILDKYGDDVKAIGVYGSLGRQTD
                     GPYSDIEMMCVMSTEEAEFSHEWTTGEWKVEVNFDSEEILLDYASQVESDWPLTHGQFF
                     SILPIYDSGGYLEKVYQTAKSVEAQTFHDAICALIVEELFEYAGKWRNIRVQGPTTFLP
                     SLTVQVAMAGAMLIGLHHRICYTTSASVLTEAVKQSDLPSGYDHLCQFVMSGQLSDSEK
                     LLESLENFWNGIQEWTERHGYIVDVSKRIPF"
     promoter        3083..3187
                     /label=AmpR promoter
     CDS             3188..4048
                     /codon_start=1
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVPLIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      4219..4807
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"

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