LOCUS       Exported                7075 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Integration expression vector pJM252, complete sequence.
ACCESSION   KX782329
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7075)
  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 7075)
  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 7075)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..7075
                     /organism="Integration expression vector pJM252"
                     /mol_type="other DNA"
                     /db_xref="taxon:1901986"
     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(902..1984)
                     /codon_start=1
                     /gene="lacI"
                     /product="transcriptional repressor LacI"
                     /label=lacI
                     /note="derived from Escherichia coli"
                     /protein_id="AOR07792.1"
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     gene            complement(902..1984)
                     /gene="lacI"
                     /label=lacI
     promoter        complement(1985..2062)
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     regulatory      complement(2013..2048)
                     /regulatory_class="promoter"
                     /note="Escherichia coli lacIq promoter"
     regulatory      2292..2326
                     /regulatory_class="promoter"
                     /note="Escherichia coli tac promoter"
     promoter        2292..2320
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     misc_feature    2327..2347
                     /label=lacO operator
                     /note="lacO operator"
     protein_bind    2328..2344
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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    2362..2415
                     /note="MCS; multiple cloning site"
     primer_bind     complement(2397..2413)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      complement(2438..2457)
                     /regulatory_class="promoter"
                     /note="bacteriophage T7 promoter"
     promoter        complement(2439..2457)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      2516..2540
                     /regulatory_class="terminator"
                     /note="bacteriophage T4 terminator"
     misc_feature    2631..2678
                     /note="FRT; Flp recombinase target site"
     protein_bind    complement(2631..2678)
                     /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            2878..3137
     oriT            2885..2993
                     /note="incP origin of transfer"
     CDS             complement(3152..4321)
                     /codon_start=1
                     /gene="int"
                     /product="bacteriophage CTX integrase"
                     /label=int
                     /protein_id="AOR07793.1"
                     /translation="MADGVEVRGKRIRIYFRYQGELCRESIPGDATPENIANAERLAGI
                     INYEIKQGVFSYSRHFPDSPRVKSNTLGHYIDLWLDIKRNQIAASGFRGYTSRVETHIR
                     PRWGDSQADSIDHLDIQDWVQNTLMPKLHNKTVREIVSNLRQIFRLYRTRNRSAHDPTD
                     GIVITLPDADDPDPFTREEIDLILGTETARIGELNLAEFMIWSGPRVSEAIALAWEDVD
                     LDTGTVVFRRARVRSQYKVTKTRRSTRKVQLLAPALRALQQQAKLTRRLPPVQIEVIDR
                     DNRTRKPQRVRFVFHNSASGAAYSTSDTLRNGWWHGHLRNAGVRSRGPNQCRHTFASQM
                     LSSGIATPEWIADQMGHTSTAMIFKHYAKWISKDGPDIVGLLNQALKLS"
     gene            complement(3152..4321)
                     /gene="int"
                     /label=int
     rep_origin      4359..5032
     rep_origin      complement(4418..5006)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5828..7018)
                     /codon_start=1
                     /gene="tet"
                     /product="tetracycline resistance protein"
                     /label=tet
                     /protein_id="AOR07794.1"
                     /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
                     GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
                     VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
                     LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
                     QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
                     AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
                     AALTSLTSITGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
     gene            complement(5828..7018)
                     /gene="tet"
                     /label=tet
     promoter        complement(7066..19)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"