LOCUS       Exported                4663 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pJLS149, complete sequence.
ACCESSION   MG752896
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4663)
  AUTHORS   Stoudenmire JL, Essock-Burns T, Weathers EN, Solaimanpour S, Mrazek 
            J, Stabb EV.
  TITLE     An iterative synthetic approach to engineer a high-performing 
            PhoB-specific reporter
  JOURNAL   Appl. Environ. Microbiol. (2018) In press
  PUBMED    29752265
REFERENCE   2  (bases 1 to 4663)
  AUTHORS   Stoudenmire JL, Stabb EV.
  TITLE     Direct Submission
  JOURNAL   Submitted (04-JAN-2018) Microbiology, University of Georgia, 120 
            Cedar Street, Athens, GA 30605, USA
REFERENCE   3  (bases 1 to 4663)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..4663
                     /organism="Cloning vector pJLS149"
                     /mol_type="other DNA"
                     /db_xref="taxon:2212377"
     CDS             62..535
                     /codon_start=1
                     /product="trimethoprim resistance protein"
                     /label=trimethoprim resistance protein
                     /note="Drf1"
                     /protein_id="AWO14315.1"
                     /translation="MKLSLMVAISKNGVIGNGPDIPWSAKGEQLLFKAITYNQWLLVGR
                     KTFESMGALPNRKYAVVTRSSFTSDNENVLIFPSIKDALTNLKKITDHVIVSGGGEIYK
                     SLIDQVDTLHISTIDIEPEGDVYFPEIPSNFRPVFTQDFASNINYSYQIWQKG"
     CDS             complement(578..1288)
                     /codon_start=1
                     /product="red florescent protein MCherry"
                     /label=red florescent protein MCherry
                     /protein_id="AWO14317.1"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     1390..1406
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(1485..2567)
                     /codon_start=1
                     /product="Lac repressor protein"
                     /label=Lac repressor protein
                     /note="LaqIQ"
                     /protein_id="AWO14316.1"
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     promoter        complement(2568..2645)
                     /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."
     protein_bind    2747..2763
                     /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)."
     protein_bind    2780..2796
                     /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)."
     primer_bind     complement(2860..2876)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(2951..2969)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      2990..3378
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     oriT            4362..4471
                     /note="incP origin of transfer"