LOCUS       Exported                6058 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Integration expression vector pJM101, complete sequence.
ACCESSION   KX782328
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6058)
  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 6058)
  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 6058)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..6058
                     /organism="Integration expression vector pJM101"
                     /mol_type="other DNA"
                     /db_xref="taxon:1901984"
     mobile_element  382..580
                     /mobile_element_type="transposon:Tn7R"
                     /note="right end of Tn7"
     primer_bind     585..601
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      629..723
                     /regulatory_class="terminator"
                     /note="bacteriophage lambda T0 terminator"
     terminator      629..723
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     regulatory      820..917
                     /regulatory_class="terminator"
                     /note="Escherichia coli rrnB T1 terminator"
     terminator      826..912
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     misc_feature    958..1005
                     /note="FRT; Flp recombinase target site"
     protein_bind    958..1005
                     /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)."
     CDS             complement(1135..1668)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamicin acetyltransferase"
                     /label=aacC1
                     /protein_id="AOR07789.1"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     gene            complement(1135..1668)
                     /gene="aacC1"
                     /label=aacC1
     promoter        complement(1857..1885)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     misc_feature    1926..1973
                     /note="FRT; Flp recombinase target site"
     protein_bind    1926..1973
                     /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)."
     CDS             complement(2050..3132)
                     /codon_start=1
                     /gene="lacI"
                     /product="transcriptional repressor LacI"
                     /label=lacI
                     /note="derived from Escherichia coli"
                     /protein_id="AOR07790.1"
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     gene            complement(2050..3132)
                     /gene="lacI"
                     /label=lacI
     promoter        complement(3133..3210)
                     /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(3161..3196)
                     /regulatory_class="promoter"
                     /note="Escherichia coli lacIq promoter"
     regulatory      3440..3474
                     /regulatory_class="promoter"
                     /note="Escherichia coli tac promoter"
     promoter        3440..3468
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     misc_feature    3475..3495
                     /label=lacO operator
                     /note="lacO operator"
     protein_bind    3476..3492
                     /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    3510..3566
                     /note="MCS; multiple cloning site"
     mobile_element  3575..3740
                     /mobile_element_type="transposon:Tn7L"
                     /note="left end of Tn7"
     oriT            3892..4159
     oriT            3899..4007
                     /note="incP origin of transfer"
     rep_origin      4180..4853
     rep_origin      complement(4239..4827)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4998..5858)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=bla
                     /protein_id="AOR07791.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(4998..5858)
                     /gene="bla"
                     /label=bla
     promoter        complement(5859..5963)
                     /gene="bla"
                     /label=AmpR promoter