LOCUS       Exported                6047 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pUC18T-mini-Tn7T-LAC-Gm, complete sequence.
ACCESSION   EU240902
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6047)
  AUTHORS   Choi KH, Mima T, Casart Y, Rholl D, Kumar A, Beacham IR, Schweizer 
            HP.
  TITLE     Genetic tools for select-agent-compliant manipulation of 
            Burkholderia pseudomallei
  JOURNAL   Appl. Environ. Microbiol. 74 (4), 1064-1075 (2008)
  PUBMED    18156318
REFERENCE   2  (bases 1 to 6047)
  AUTHORS   Choi K-H., Schweizer HP.
  TITLE     Genetic tools for Pseudomonas and its related bacteria
  JOURNAL   Unpublished
REFERENCE   3  (bases 1 to 6047)
  AUTHORS   Choi K-H., Schweizer HP.
  TITLE     Direct Submission
  JOURNAL   Submitted (23-OCT-2007) Microbiology, Immunology and Pathology, 
            Colorado State University, 1682 Campus delivery, Ft. Collins, CO 
            80523, USA
REFERENCE   4  (bases 1 to 6047)
  AUTHORS   Choi K-H., Schweizer HP.
  TITLE     Direct Submission
  JOURNAL   Submitted (25-MAY-2010) Microbiology, Immunology and Pathology, 
            Colorado State University, 1682 Campus delivery, Ft. Collins, CO 
            80523, USA
REFERENCE   5  (bases 1 to 6047)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     On May 25, 2010 this sequence version replaced EU240902.1.
FEATURES             Location/Qualifiers
     source          1..6047
                     /organism="Expression vector pUC18T-mini-Tn7T-LAC-Gm"
                     /mol_type="other DNA"
                     /db_xref="taxon:483254"
     repeat_region   382..580
                     /label=right end of Tn7
                     /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      629..723
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     regulatory      820..917
                     /regulatory_class="terminator"
                     /note="Escherichia coli rrnB T1"
     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="gentamycin acetyl transferase"
                     /label=aacC1
                     /protein_id="ABX52091.1"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     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)."
     gene            complement(2016..3164)
                     /gene="lacI"
                     /label=lacI
     CDS             complement(2016..3098)
                     /codon_start=1
                     /gene="lacI"
                     /product="Lac repressor"
                     /label=lacI
                     /protein_id="ABX52092.1"
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPSTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     promoter        complement(3099..3176)
                     /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(3134..3164)
                     /regulatory_class="promoter"
                     /gene="lacI"
                     /note="lacI promoter region with lacIQ promoter-up"
     promoter        3406..3434
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     regulatory      3406..3433
                     /regulatory_class="promoter"
                     /note="tac"
     protein_bind    3440..3460
                     /label=LacI binding site
                     /bound_moiety="LacI"
                     /note="lac operator"
     protein_bind    3442..3458
                     /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    3475..3555
                     /note="MCS; multiple cloning site"
     repeat_region   3564..3729
                     /label=left end of Tn7
                     /note="left end of Tn7"
     mobile_element  3564..3729
                     /mobile_element_type="transposon:Tn7"
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      3881..4148
                     /note="oriT; origin of conjugal transfer"
     oriT            3888..3996
                     /note="incP origin of transfer"
     rep_origin      complement(4228..4816)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4987..5847)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=bla
                     /protein_id="ABX52093.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(4987..5847)
                     /gene="bla"
                     /label=bla
     promoter        complement(5848..5952)
                     /gene="bla"
                     /label=AmpR promoter