LOCUS       Exported                7960 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Reporter vector pGlowRed, complete sequence.
ACCESSION   HM049183
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7960)
  AUTHORS   Krol JE, Rogers LM, Krone SM, Top EM.
  TITLE     Dual reporter system for in situ detection of plasmid transfer under
            aerobic and anaerobic conditions
  JOURNAL   Appl. Environ. Microbiol. 76 (13), 4553-4556 (2010)
  PUBMED    20453134
REFERENCE   2  (bases 1 to 7960)
  AUTHORS   Krol JE.
  TITLE     Direct Submission
  JOURNAL   Submitted (31-MAR-2010) Biological Sciences, University of Idaho, 
            P.O. Box 445031, Moscow, ID 83844, USA
REFERENCE   3  (bases 1 to 7960)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7960
                     /organism="Reporter vector pGlowRed"
                     /mol_type="other DNA"
                     /db_xref="taxon:797074"
     CDS             2..433
                     /codon_start=1
                     /gene="EcFbFP"
                     /product="Evoglow"
                     /label=EcFbFP
                     /protein_id="ADJ00079.1"
                     /translation="MASFQSFGIPGQLEVIKKALDHVRVGVVITDPALEDNPIVYVNQG
                     FVQMTGYETEEILGKNARFLQGKHTDPAEVDNIRTALQNKEPVTVQIQNYKKDGTMFWN
                     ELNIDPMEIEDKTYFVGIQNDITKQKEYEKLLEHHHHHH"
     gene            2..433
                     /gene="EcFbFP"
                     /label=EcFbFP
     CDS             413..430
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      497..544
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1417..2079)
                     /codon_start=1
                     /product="replication protein for the broad-host-range 
                     plasmid pBBR1 from Bordetella bronchiseptica"
                     /label=pBBR1 Rep
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
     rep_origin      2080..2849
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     promoter        4242..4344
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             4345..5364
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=cat
                     /protein_id="ADJ00076.1"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQFLCMRPIRKPPL
                     PARWPIHYSHAPRPPEGADWVEGSQGHRSRSRCLMSELTYINCVALTARFPVGKPVVPA
                     ALMNRPTRGERRFAYWAPGWFFFSPVRRATADCPSPPGPERVAASGPRWFAPAGENPV"
     gene            4345..5364
                     /gene="cat"
                     /label=cat
     CDS             complement(5430..6107)
                     /codon_start=1
                     /gene="dsRed"
                     /product="red fluorescent protein"
                     /label=dsRed
                     /protein_id="ADJ00077.1"
                     /translation="MRSSKNVIKEFMRFKVRMEGTVNGHEFEIEGEGEGRPYEGHNTVK
                     LKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGCFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGEIHK
                     ALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDSKLDITSHNEDYTIVEQYERTEGRHH
                     LFL"
     gene            complement(5430..6107)
                     /gene="dsRed"
                     /label=dsRed
     RBS             6114..6125
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     promoter        complement(6175..6193)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     regulatory      complement(6194..6225)
                     /regulatory_class="promoter"
                     /note="pLac promoter"
     primer_bind     complement(6211..6227)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    6235..6251
                     /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)."
     promoter        complement(6259..6289)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6304..6325
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             complement(6696..7490)
                     /codon_start=1
                     /gene="KmR"
                     /product="kanamycin phosphotransferase"
                     /label=KmR
                     /protein_id="ADJ00078.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     gene            complement(6696..7490)
                     /gene="KmR"
                     /label=KmR
     promoter        7874..7892
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      7874..7889
                     /regulatory_class="promoter"
                     /note="T7 promoter"
     protein_bind    7893..7917
                     /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)."
     RBS             7932..7954
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"