pJM253 vector (V005294)

Basic Vector Information

Vector Name:
pJM253
Antibiotic Resistance:
Tetracycline
Length:
7711 bp
Type:
Integration expression vector
Replication origin:
ori
Source/Author:
Meisner J, Goldberg JB.
Promoter:
rhaB

pJM253 vector Map

pJM2537711 bp3006009001200150018002100240027003000330036003900420045004800510054005700600063006600690072007500tet promoterT3 promoterrhaRrhaSrhaB promoterKS primerT7 promoterFRToriToriTcR

pJM253 vector Sequence

LOCUS       40924_26306        7711 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Integration expression vector pJM253, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7711)
  AUTHORS   Meisner J, Goldberg JB.
  TITLE     Escherichia coli rhaSR-PrhaBAD inducible promoter system allows 
            tightly controlled gene expression over a wide range in Pseudomonas 
            aeruginosa
  JOURNAL   Appl. Environ. Microbiol. (2016) In press
  PUBMED    27613678
REFERENCE   2  (bases 1 to 7711)
  AUTHORS   Meisner J.
  TITLE     Direct Submission
  JOURNAL   Submitted (26-AUG-2016) Biochemistry, Emory University School of 
            Medicine, 1510 Clifton Road NE, Rollins Research Center, room G223, 
            Atlanta, GA 30322, USA
REFERENCE   3  (bases 1 to 7711)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7711)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Appl. 
            Environ. Microbiol. (2016) In press"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (26-AUG-2016) Biochemistry, Emory University School of Medicine, 
            1510 Clifton Road NE, Rollins Research Center, room G223, Atlanta, 
            GA 30322, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..7711
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        827..845
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     CDS             complement(968..1813)
                     /codon_start=1
                     /label=rhaR
                     /note="transcriptional activator of rhaR and rhaS"
                     /translation="VAHQLKLLKDDFFASDQQAVAVADRYPQDVFAEHTHDFCELVIVW
                     RGNGLHVLNDRPYRITRGDLFYIHADDKHSYASVNDLVLQNIIYCPERLKLNLDWQGAI
                     PGFNASAGQPHWRLGSMGMAQARQVIGQLEHESSQHVPFANEMAELLFGQLVMLLNRHR
                     YTSDSLPPTSSETLLDKLITRLAASLKSPFALDKFCDEASCSERVLRQQFRQQTGMTIN
                     QYLRQVRVCHAQYLLQHSRLLISDISTECGFEDSNYFSVVFTRETGMTPSQWRHLNSQK
                     D"
     CDS             complement(1890..2723)
                     /codon_start=1
                     /label=rhaS
                     /note="positive regulator of the rhaB promoter"
                     /translation="MTVLHSVDFFPSGNASVAIEPRLPQADFPEHHHDFHEIVIVEHGT
                     GIHVFNGQPYTITGGTVCFVRDHDRHLYEHTDNLCLTNVLYRSPDRFQFLAGLNQLLPQ
                     ELDGQYPSHWRVNHSVLQQVRQLVAQMEQQEGENDLPSTASREILFMQLLLLLRKSSLQ
                     ENLENSASRLNLLLAWLEDHFADEVNWDAVADQFSLSLRTLHRQLKQQTGLTPQRYLNR
                     LRLMKARHLLRHSEASVTDIAYRCGFSDSNHFSTLFRREFNWSPRDIRQGRDGFLQ"
     promoter        2869..2987
                     /label=rhaB promoter
                     /note="promoter of the E. coli rhaBAD operon, conferring
                     tight induction with L-rhamnose and repression with 
                     D-glucose in the presence of RhaR and RhaS (Giacalone et 
                     al., 2006)"
     primer_bind     complement(3033..3049)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(3075..3093)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    complement(3267..3314)
                     /label=FRT
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     oriT            complement(3521..3629)
                     /direction=LEFT
                     /label=oriT
                     /note="incP origin of transfer"
     rep_origin      complement(5054..5642)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6467..7654)
                     /codon_start=1
                     /label=TcR
                     /note="tetracycline efflux protein"
                     /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
                     GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
                     VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
                     LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
                     QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
                     AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
                     AALTSLTSITGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
     promoter        complement(join(7702..7711,1..19))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein
                     gene"

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