LOCUS       Exported                7626 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Shuttle vector pEX1.8 plasmid pEX1.8, complete sequence.
ACCESSION   JQ342676
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7626)
  AUTHORS   Pearson JP, Pesci EC, Iglewski BH.
  TITLE     Roles of Pseudomonas aeruginosa las and rhl quorum-sensing systems 
            in control of elastase and rhamnolipid biosynthesis genes
  JOURNAL   J. Bacteriol. 179 (18), 5756-5767 (1997)
  PUBMED    9294432
REFERENCE   2  (bases 1 to 7626)
  AUTHORS   Schwingel JM, Iglewski BH.
  TITLE     pEX1.8, Pseudomonas aeruginosa/E. coli shuttle vector
  JOURNAL   Unpublished
REFERENCE   3  (bases 1 to 7626)
  AUTHORS   Schwingel JM, Iglewski BH.
  TITLE     Direct Submission
  JOURNAL   Submitted (02-JAN-2012) Department of Microbiology and Immunology, 
            University of Rochester, 601 Elmwood Ave., Box 672, Rochester, NY 
            14642, USA
REFERENCE   4  (bases 1 to 7626)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7626
                     /organism="Shuttle vector pEX1.8"
                     /plasmid="pEX1.8"
                     /mol_type="other DNA"
                     /note="Pseudomonas aeruginosa/E. coli shuttle vector with 
                     pTac promoter"
                     /db_xref="taxon:1223316"
     rep_origin      1161..3044
                     /note="Pseudomonas aeruginosa stabilization fragment from 
                     pRO1614; allows for plasmid replication"
     rep_origin      1813..2164
                     /label=pRO1600 oriV
                     /note="broad-host-range origin of replication from 
                     Pseudomonas aeruginosa plasmid pRO1600; requires the 
                     pRO1600 Rep protein for replication (West et al., 1994)"
     CDS             2178..3011
                     /codon_start=1
                     /product="replication protein for the broad-host-range 
                     plasmid pRO1600 from Pseudomonas aeruginosa "
                     /label=pRO1600 Rep
                     /translation="MASPPMVYKSNALVEAAYRLSVQEQRIVLACISQVKRSEPVTDEV
                     MYSVTAEDIATMAGVPIESSYNQLKEAALRLKRREVRLTQEPNGKGKRPSVMITGWVQT
                     IIYREGEGRVELRFTKDMLPYLTELTKQFTKYALADVAKMDSTHAIRLYELLMQWDSIG
                     QREIEIDQLRKWFQLEGRYPSIKDFKLRVLDPAVTQINEHSPLQVEWAQRKTGRKVTHL
                     LFSFGPKKPAKAVGKAPAKRKAGKISDAEIAKQARPGETWEAARARLTQMPLDLA"
     promoter        4120..4148
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     regulatory      4120..4147
                     /regulatory_class="promoter"
                     /note="pTac promoter"
     protein_bind    4156..4172
                     /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_structure  4189..4224
                     /note="multiple cloning site; MCS; contains 
                     EcoRI-XmaI/SmaI-PstI-HindIII"
     terminator      4427..4513
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4605..4632
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        4651..4742
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4743..5603
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=bla
                     /note="confers ampicillin resistance; derived from pBR322"
                     /protein_id="AFQ23787.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            4743..5603
                     /gene="bla"
                     /label=bla
     rep_origin      5774..6362
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      5948..6178
                     /label=Escherichia coli colE1 origin
                     /note="Escherichia coli colE1 origin"
     misc_feature    6548..6688
                     /label=bom
                     /note="basis of mobility region from pBR322"
     promoter        6864..6941
                     /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."