LOCUS       Exported                8603 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Expression vector pBBRKW2HTsLsM, complete sequence.
ACCESSION   JN565028
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8603)
  AUTHORS   Faries KM, Kressel LL, Wander MJ, Holten D, Laible PD, Kirmaier C, 
            Hanson DK.
  TITLE     High throughput engineering to revitalize a vestigial electron 
            transfer pathway in bacterial photosynthetic reaction centers
  JOURNAL   J. Biol. Chem. 287 (11), 8507-8514 (2012)
  PUBMED    22247556
REFERENCE   2  (bases 1 to 8603)
  AUTHORS   Kressel L, Wander MJ, Hanson DK, Laible PD.
  TITLE     Direct Submission
  JOURNAL   Submitted (08-AUG-2011) Biosciences, Argonne National Laboratory, 
            9700 S. Cass Ave., Argonne, IL 60439, USA
REFERENCE   3  (bases 1 to 8603)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8603
                     /organism="Expression vector pBBRKW2HTsLsM"
                     /mol_type="other DNA"
                     /db_xref="taxon:1128098"
     rep_origin      1023..1792
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     CDS             1793..2455
                     /codon_start=1
                     /gene="rep"
                     /product="Rep"
                     /function="required for plasmid replication"
                     /label=rep
                     /protein_id="AEW50175.1"
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
     gene            1793..2455
                     /gene="rep"
                     /label=rep
     primer_bind     3162..3178
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        3188..3206
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             complement(4132..4368)
                     /codon_start=1
                     /gene="pufX"
                     /product="intrinsic membrane protein PufX"
                     /label=pufX
                     /protein_id="AEW50176.1"
                     /translation="MSMFDKPFDYENGSKFEMGIWIGRQMAYGAFLGSIPFLLGLGLVL
                     GSYGLGLMLPERAHQAPSPYTTEVVVQHATEVV"
     gene            complement(4132..4368)
                     /gene="pufX"
                     /label=pufX
     CDS             complement(4381..5325)
                     /codon_start=1
                     /gene="pufM"
                     /product="photosynthetic reaction center subunit M"
                     /label=pufM
                     /protein_id="AEW50177.1"
                     /translation="MAEYQNFFNQVQVAGAPEMGLKEDVDTFERTPAGMFNILGWMGNA
                     QIGPIYLGIAGTVSLAFGAAWFFTIGVWYWYQAGFDPFIFMRDLFFFSLEPPPAEYGLA
                     IAPLKQGGVWQIASLFMAISVIAWWVRVYTRADQLGMGKHMAWAFLSAIWLWSVLGFWR
                     PILMGSWSVAPPYGIFSHLDWTNQFSLDHGNLFYNPFHGLSIAALYGSALLFAMHGATI
                     LAVTRFGGERELEQIVDRGTASERAALFWRWTMGFNATMEGIHRWAIWMAVMVTLTGGI
                     GILLSGTVVDNWYVWAQVHGYAPVTPHHHHHHH"
     gene            complement(4381..5325)
                     /gene="pufM"
                     /label=pufM
     CDS             complement(4384..4404)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=7xHis
                     /translation="HHHHHHH"
     regulatory      4542..4551
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             complement(5318..6166)
                     /codon_start=1
                     /gene="pufL"
                     /product="photosynthetic reaction center subunit L"
                     /label=pufL
                     /protein_id="AEW50178.1"
                     /translation="MALLSFERKYRVPGGTLIGGSLFDFWVGPFYVGFFGVTTIFFATL
                     GFLLILWGAAMQGTWNPQLISIFPPPVENGLNVAALDKGGLWQVITVCVTGAFCSWALR
                     EVEICRKLGIGFHIPVAFSMAIFAYLTLVVIRPMMMGSWGYAFPYGIWTHLDWVSNTGY
                     TYGNFHYNPFHMLGISLFFTTAWALAMHGALVLSAANPVKGKTMRTPDHEDTYFRDLMG
                     YSVGTLGIHRLGLLLALNAVFWSACCMLVSGTIYFDLWSDWWYWWVNMPFWADMAGGIN
                     G"
     gene            complement(5318..6166)
                     /gene="pufL"
                     /label=pufL
     CDS             complement(6299..6475)
                     /codon_start=1
                     /gene="pufA"
                     /product="light-harvesting protein B-875 subunit alpha"
                     /label=pufA
                     /protein_id="AEW50179.1"
                     /translation="MSKFYKIWLVFDPRRVFVAQGVFLFLLAVLIRLILLSTPAFNWLT
                     VATAKHGYVAAAQ"
     gene            complement(6299..6475)
                     /gene="pufA"
                     /label=pufA
     CDS             complement(6489..6638)
                     /codon_start=1
                     /gene="pufB"
                     /product="light-harvesting protein B-875 subunit beta"
                     /label=pufB
                     /protein_id="AEW50180.1"
                     /translation="MADKNDLSFTGLTDEQAQELHAVYMSGLSAFIAVAVLAHLAVMIW
                     RPWF"
     gene            complement(6489..6638)
                     /gene="pufB"
                     /label=pufB
     promoter        complement(6802..6820)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6841..6857)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    6865..6881
                     /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(6889..6919)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6934..6955
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             complement(7356..8150)
                     /codon_start=1
                     /gene="kanR"
                     /product="kanamycin resistance protein"
                     /label=kanR
                     /protein_id="AEW50181.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     gene            complement(7356..8150)
                     /gene="kanR"
                     /label=kanR
     CDS             complement(8404..799)
                     /codon_start=1
                     /gene="mob"
                     /product="Mob"
                     /function="required for plasmid mobilization"
                     /label=mob
                     /protein_id="AEW50174.1"
                     /translation="MAAYAIMRCKKLAKMGNVAASLKHAYRERETPNADASRTPENEHW
                     AASSTDEAMGRLRELLPEKRRKDAVLAVEYVMTASPEWWKSASQEQQAAFFEKAHKWLA
                     DKYGADRIVTASIHRDETSPHMTAFVVPLTQDGRLSAKEFIGNKAQMTRDQTTFAAAVA
                     DLGLQRGIEGSKARHTRIQAFYEALERPPVGHVTISPQAVEPRAYAPQGLAEKLGISKR
                     VETPEAVADRLTKAVRQGYEPALQAAAGAREMRKKADQAQETARDLRERLKPVLDALGP
                     LNRDMQAKAAAIIKAVGEKLLTEQREVQRQKQAQRQQERGRAHFPEKCHLG"
     gene            complement(8404..799)
                     /gene="mob"
                     /label=mob