LOCUS       Exported                6797 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Integration expression vector pJM251, complete sequence.
ACCESSION   KX787912
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6797)
  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 6797)
  AUTHORS   Meisner J.
  TITLE     Direct Submission
  JOURNAL   Submitted (29-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 6797)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..6797
                     /organism="Integration expression vector pJM251"
                     /mol_type="other DNA"
                     /db_xref="taxon:1899432"
     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(875..1753)
                     /codon_start=1
                     /gene="araC"
                     /product="transcriptional regulator AraC"
                     /label=araC
                     /note="derived from Escherichia coli"
                     /protein_id="AOO86786.1"
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     gene            complement(875..1753)
                     /gene="araC"
                     /label=araC
     promoter        1780..2064
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     misc_feature    1782..1799
                     /label=araO2 operator
                     /note="araO2 operator"
     regulatory      complement(1904..1932)
                     /regulatory_class="promoter"
                     /note="Escherichia coli araC promoter"
     misc_feature    1940..1961
                     /label=araO1 operator
                     /note="araO1 operator"
     misc_feature    1993..2009
                     /label=araI1 operator
                     /note="araI1 operator"
     misc_feature    2014..2030
                     /label=araI2 operator
                     /note="araI2 operator"
     regulatory      2027..2065
                     /regulatory_class="promoter"
                     /note="Escherichia coli araBAD promoter"
     misc_feature    2084..2131
                     /note="MCS; multiple cloning site"
     primer_bind     complement(2119..2135)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      complement(2160..2179)
                     /regulatory_class="promoter"
                     /note="bacteriophage T7 promoter"
     promoter        complement(2161..2179)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      2238..2262
                     /regulatory_class="terminator"
                     /note="bacteriophage T4 terminator"
     misc_feature    2353..2400
                     /note="FRT; Flp recombinase target site"
     protein_bind    complement(2353..2400)
                     /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            2600..2859
     oriT            2607..2715
                     /note="incP origin of transfer"
     CDS             complement(2874..4043)
                     /codon_start=1
                     /gene="int"
                     /product="bacteriophage CTX integrase"
                     /label=int
                     /protein_id="AOO86787.1"
                     /translation="MADGVEVRGKRIRIYFRYQGELCRESIPGDATPENIANAERLAGI
                     INYEIKQGVFSYSRHFPDSPRVKSNTLGHYIDLWLDIKRNQIAASGFRGYTSRVETHIR
                     PRWGDSQADSIDHLDIQDWVQNTLMPKLHNKTVREIVSNLRQIFRLYRTRNRSAHDPTD
                     GIVITLPDADDPDPFTREEIDLILGTETARIGELNLAEFMIWSGPRVSEAIALAWEDVD
                     LDTGTVVFRRARVRSQYKVTKTRRSTRKVQLLAPALRALQQQAKLTRRLPPVQIEVIDR
                     DNRTRKPQRVRFVFHNSASGAAYSTSDTLRNGWWHGHLRNAGVRSRGPNQCRHTFASQM
                     LSSGIATPEWIADQMGHTSTAMIFKHYAKWISKDGPDIVGLLNQALKLS"
     gene            complement(2874..4043)
                     /gene="int"
                     /label=int
     rep_origin      4081..4754
     rep_origin      complement(4140..4728)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5550..6740)
                     /codon_start=1
                     /gene="tet"
                     /product="tetracycline resistance protein"
                     /label=tet
                     /protein_id="AOO86788.1"
                     /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
                     GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
                     VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
                     LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
                     QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
                     AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
                     AALTSLTSITGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
     gene            complement(5550..6740)
                     /gene="tet"
                     /label=tet
     promoter        complement(6788..19)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"