LOCUS       Exported                7711 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Integration expression vector pJM253, complete sequence.
ACCESSION   KX782327
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)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..7711
                     /organism="Integration expression vector pJM253"
                     /mol_type="other DNA"
                     /db_xref="taxon:1901987"
     misc_feature    238..285
                     /note="FRT; Flp recombinase target site"
     protein_bind    complement(238..285)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     misc_feature    495..521
                     /note="attP; bacteriophage CTX attachment site"
     regulatory      745..769
                     /regulatory_class="terminator"
                     /note="bacteriophage T4 terminator"
     regulatory      827..846
                     /regulatory_class="promoter"
                     /note="bacteriophage T3 promoter"
     promoter        827..845
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     CDS             complement(965..1813)
                     /codon_start=1
                     /gene="rhaR"
                     /product="transcriptional activator RhaR"
                     /label=rhaR
                     /note="derived from Escherichia coli"
                     /protein_id="AOR07786.1"
                     /translation="MAHQLKLLKDDFFASDQQAVAVADRYPQDVFAEHTHDFCELVIVW
                     RGNGLHVLNDRPYRITRGDLFYIHADDKHSYASVNDLVLQNIIYCPERLKLNLDWQGAI
                     PGFNASAGQPHWRLGSMGMAQARQVIGQLEHESSQHVPFANEMAELLFGQLVMLLNRHR
                     YTSDSLPPTSSETLLDKLITRLAASLKSPFALDKFCDEASCSERVLRQQFRQQTGMTIN
                     QYLRQVRVCHAQYLLQHSRLLISDISTECGFEDSNYFSVVFTRETGMTPSQWRHLNSQK
                     D"
     gene            complement(965..1813)
                     /gene="rhaR"
                     /label=rhaR
     CDS             complement(1887..2723)
                     /codon_start=1
                     /gene="rhaS"
                     /product="transcriptional activator RhaS"
                     /label=rhaS
                     /note="derived from Escherichia coli"
                     /protein_id="AOR07785.1"
                     /translation="MTVLHSVDFFPSGNASVAIEPRLPQADFPEHHHDFHEIVIVEHGT
                     GIHVFNGQPYTITGGTVCFVRDHDRHLYEHTDNLCLTNVLYRSPDRFQFLAGLNQLLPQ
                     ELDGQYPSHWRVNHSVLQQVRQLVAQMEQQEGENDLPSTASREILFMQLLLLLRKSSLQ
                     ENLENSASRLNLLLAWLEDHFADEVNWDAVADQFSLSLRTLHRQLKQQTGLTPQRYLNR
                     LRLMKARHLLRHSEASVTDIAYRCGFSDSNHFSTLFRREFNWSPRDIRQGRDGFLQ"
     gene            complement(1887..2723)
                     /gene="rhaS"
                     /label=rhaS
     regulatory      complement(2748..2783)
                     /regulatory_class="promoter"
                     /note="Escherichia coli rhaSR promoter"
     misc_feature    2780..2796
                     /label=rhaI4 operator
                     /note="rhaI4 operator"
     misc_feature    2814..2830
                     /label=rhaI3 operator
                     /note="rhaI3 operator"
     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)"
     misc_feature    2906..2922
                     /label=rhaI1 operator
                     /note="rhaI1 operator"
     misc_feature    2939..2955
                     /label=rhaI2 operator
                     /note="rhaI2 operator"
     regulatory      2952..2987
                     /regulatory_class="promoter"
                     /note="Escherichia coli rhaBAD promoter"
     misc_feature    2998..3045
                     /note="MCS; multiple cloning site"
     primer_bind     complement(3033..3049)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      complement(3074..3093)
                     /regulatory_class="promoter"
                     /note="bacteriophage T7 promoter"
     promoter        complement(3075..3093)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      3152..3176
                     /regulatory_class="terminator"
                     /note="bacteriophage T4 terminator"
     misc_feature    3267..3314
                     /note="FRT; Flp recombinase target site"
     protein_bind    complement(3267..3314)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     oriT            3514..3773
     oriT            3521..3629
                     /note="incP origin of transfer"
     CDS             complement(3788..4957)
                     /codon_start=1
                     /gene="int"
                     /product="bacteriophage CTX integrase"
                     /label=int
                     /protein_id="AOR07788.1"
                     /translation="MADGVEVRGKRIRIYFRYQGELCRESIPGDATPENIANAERLAGI
                     INYEIKQGVFSYSRHFPDSPRVKSNTLGHYIDLWLDIKRNQIAASGFRGYTSRVETHIR
                     PRWGDSQADSIDHLDIQDWVQNTLMPKLHNKTVREIVSNLRQIFRLYRTRNRSAHDPTD
                     GIVITLPDADDPDPFTREEIDLILGTETARIGELNLAEFMIWSGPRVSEAIALAWEDVD
                     LDTGTVVFRRARVRSQYKVTKTRRSTRKVQLLAPALRALQQQAKLTRRLPPVQIEVIDR
                     DNRTRKPQRVRFVFHNSASGAAYSTSDTLRNGWWHGHLRNAGVRSRGPNQCRHTFASQM
                     LSSGIATPEWIADQMGHTSTAMIFKHYAKWISKDGPDIVGLLNQALKLS"
     gene            complement(3788..4957)
                     /gene="int"
                     /label=int
     rep_origin      4995..5668
     rep_origin      complement(5054..5642)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6464..7654)
                     /codon_start=1
                     /gene="tet"
                     /product="tetracycline resistance protein"
                     /label=tet
                     /protein_id="AOR07787.1"
                     /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
                     GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
                     VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
                     LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
                     QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
                     AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
                     AALTSLTSITGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
     gene            complement(6464..7654)
                     /gene="tet"
                     /label=tet
     promoter        complement(7702..19)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"