LOCUS       Exported               10546 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Broad-host-range expression vector pQLICE, complete sequence.
ACCESSION   EF189157
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10546)
  AUTHORS   Harms K, Schon V, Kickstein E, Wackernagel W.
  TITLE     The RecJ DNase strongly suppresses genomic integration of short but 
            not long foreign DNA fragments by homology-facilitated illegitimate 
            recombination during transformation of Acinetobacter baylyi
  JOURNAL   Mol. Microbiol. 64 (3), 691-702 (2007)
  PUBMED    17462017
REFERENCE   2  (bases 1 to 10546)
  AUTHORS   Kickstein E, Harms K, Wackernagel W.
  TITLE     Deletions of recBCD or recD influence genetic transformation 
            differently and are lethal together with a recJ deletion in 
            Acinetobacter baylyi
  JOURNAL   Microbiology (Reading, Engl.) 153 (PT 7), 2259-2270 (2007)
  PUBMED    17600070
REFERENCE   3  (bases 1 to 10546)
  AUTHORS   Huelter N.
  TITLE     Direct Submission
  JOURNAL   Submitted (15-DEC-2006) Genetics, Department of Biology and 
            Environmental Sciences, Carl von Ossietzky University of Oldenburg, 
            Carl-von-Ossietzky-Strasse 9-11, Oldenburg D-26111, Germany
REFERENCE   4  (bases 1 to 10546)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..10546
                     /organism="Broad host range expression vector pQLICE"
                     /mol_type="other DNA"
                     /note="incompatibility group Q; mobilizable; derivative of 
                     plasmid RSF1010"
                     /db_xref="taxon:417992"
     CDS             63..866
                     /codon_start=1
                     /gene="strA"
                     /product="streptomycin resistance protein A"
                     /label=strA
                     /protein_id="ABM30607.1"
                     /translation="MNRTNIFFGESHSDWLPVRGGESGDFVFRRGDGHAFAKIAPASRR
                     GELAGERDRLIWLKGRGVACPEVINWQEEQEGACLVITAIPGVPAADLSGADLLKAWPS
                     MGQQLGAVHSLSVDQCPFERRLSRMFGRAVDVVSRNAVNPDFLPDEDKSTPLHDLLARV
                     ERELPVRLDQERTDMVVCHGDPCMPNFMVDPKTLQCTGLIDLGRLGTADRYADLALMIA
                     NAEENWAAPDEAERAFAVLFNVLGIEAPDRERLAFYLRLDPLTWG"
     gene            63..866
                     /gene="strA"
                     /label=strA
     CDS             866..1702
                     /codon_start=1
                     /gene="strB"
                     /product="streptomycin resistance protein B"
                     /label=strB
                     /protein_id="ABM30608.1"
                     /translation="MFMPPVFPAHWHVSQPVLIADTFSSLVWKVSLPDGTPAIVKGLKP
                     IEDIADELRGADYLVWRNGRGAVRLLGRENNLMLLEYAGERMLSHIVAEHGDYQATEIA
                     AELMAKLYAASEEPLPSALLPIRDRFAALFQRARDDQNAGCQTDYVHAAIIADQMMSNA
                     SELRGLHGDLHHENIMFSSRGWLVIDPVGLVGEVGFGAANMFYDPADRDDLCLDPRRIA
                     QMADAFSRALDVDPRRLLDQAYAYGCLSAAWNADGEEEQRDLAIAAAIKQVRQTSY"
     gene            866..1702
                     /gene="strB"
                     /label=strB
     misc_feature    1674..2034
                     /note="fragment; similar to transposase"
     misc_feature    2035..2050
                     /label=multiple cloning site
                     /note="multiple cloning site"
     regulatory      complement(2052..2128)
                     /regulatory_class="promoter"
                     /note="tac promoter"
     protein_bind    complement(2065..2099)
                     /label=LacI repressor protein binding site
                     /bound_moiety="LacI repressor protein"
                     /note="lac operator"
     protein_bind    2074..2090
                     /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(2098..2126)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     promoter        2360..2437
                     /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      2372..2403
                     /regulatory_class="promoter"
                     /function="lacIq promoter"
     misc_feature    2375
                     /label=lacIq point mutation
                     /note="lacIq point mutation"
     CDS             2438..3520
                     /codon_start=1
                     /gene="lacIq"
                     /product="LacI repressor protein"
                     /label=lacIq
                     /protein_id="ABM30609.1"
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPSTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     gene            2438..3520
                     /gene="lacIq"
                     /label=lacIq
     misc_feature    3897..4075
                     /note="fragment; similar to transposase"
     rep_origin      4208..4602
                     /label=RSF1010 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     RSF1010; requires the RSF1010 RepA/B/C proteins for 
                     replication (Scholz et al., 1989)"
     rep_origin      4209..4633
                     /standard_name="oriV"
                     /label=oriV
                     /note="oriV; independent of host replication functions; 
                     requires RepA/B/C; replication can proceed either uni- or 
                     bidirectionally"
     repeat_region   4212..4275
                     /rpt_unit_range=4212 .. 4231
                     /note="incompatibility region; replication-specific 
                     iterons; RepC binding sites"
     misc_feature    4416..4562
                     /note="ssiB; site of DNA priming for replication; 
                     recognized by RepB"
     misc_feature    4563..4610
                     /note="ssiA; site of DNA priming for replication; 
                     recognized by RepB"
     CDS             complement(4629..4913)
                     /codon_start=1
                     /gene="mobC"
                     /product="mobilization protein C"
                     /label=mobC
                     /protein_id="ABM30610.1"
                     /translation="MVKGSNKAADRLAKLEEQRARINAEIQRVRAREQQQERKNETRRK
                     VLVGAMILAKVNSSEWPEDRLMAAMDAYLERDHDRALFGLPPRQKDEPG"
     gene            complement(4629..4913)
                     /gene="mobC"
                     /label=mobC
     oriT            4867..5023
                     /label=required in cis for mobilization
                     /note="required in cis for mobilization"
     oriT            4944..5031
                     /label=RSF1010 oriT
                     /note="origin of transfer of the broad-host-range plasmid 
                     RSF1010 (Scholz et al., 1989)"
     regulatory      4964..4968
                     /regulatory_class="promoter"
                     /note="PI"
     protein_bind    4997..5007
                     /label=MobA binding site
                     /bound_moiety="MobA"
     repeat_region   5022..5031
                     /label=imperfect repeat A
                     /note="imperfect repeat A"
     CDS             5112..7241
                     /codon_start=1
                     /gene="mobA"
                     /product="mobilization protein A"
                     /label=mobA
                     /note="site-specific endonuclease; functions during plasmid
                     transfer; recognizes aagtgcgccct; cleaves after g at 
                     position 7 and remains covalently attached to the 5 
                     phosphate through Tyr24"
                     /protein_id="ABM30611.1"
                     /translation="MAIYHLTAKTGSRSGGQSARAKADYIQREGKYARDMDEVLHAESG
                     HMPEFVERPADYWDAADLYERANGRLFKEVEFALPVELTLDQQKALASEFAQHLTGAER
                     LPYTLAIHAGGGENPHCHLMISERINDGIERPAAQWFKRYNGKTPEKGGAQKTEALKPK
                     AWLEQTREAWADHANRALERAGHDARIDHRTLEAQGIERLPGVHLGPNVVEMEGRGIRT
                     DRADVALNIDTANAQIIDLQEYREAIDHERNRQSEEIQRHQRVSGADRTAGPEHGDTGR
                     RSPAGHEPDPAGQRGAGGGVAESPAPDRGGMGGAGQRVAGGSRRGEQRRAERPERVAGV
                     ALEAMANRDAGFHDAYGGAADRIVALARPDATDNRGRLDLAALGGPMKNDRTLQAIGRQ
                     LKAMGCERFDIGVRDATTGQMMNREWSAAEVLQNTPWLKRMNAQGNDVYIRPAEQERHG
                     LVLVDDLSEFDLDDMKAEGREPALVVETSPKNYQAWVKVADAAGGELRGQIARTLASEY
                     DADPASADSRHYGRLAGFTNRKDKHTTRAGYQPWVLLRESKGKTATAGPALVQQAGQQI
                     EQAQRQQEKARRLASLELPERQLSRHRRTALDEYRSEMAGLVKRFGDDLSKCDFIAAQK
                     LASRGRSAEEIGKAMAEASPALAERKPGHEADYIERTVSKVMGLPSVQLARAELARAPA
                     PRQRGMDRGGPDFSM"
     gene            5112..7241
                     /gene="mobA"
                     /label=mobA
     CDS             5860..6273
                     /codon_start=1
                     /gene="mobB"
                     /product="mobilization protein B"
                     /label=mobB
                     /protein_id="ABM30612.1"
                     /translation="MNAIDRVKKSRGINELAEQIEPLAQSMATLADEARQVMSQTQQAS
                     EAQAAEWLKAQRQTGAAWVELAKELREVAAEVSSAAQSARSASRGWHWKLWLTVMLASM
                     MPTVVLLIASLLLLDLTPLTTEDGSIWLRLVAR"
     gene            5860..6273
                     /gene="mobB"
                     /label=mobB
     CDS             6270..7241
                     /codon_start=1
                     /gene="repB"
                     /product="replication protein B"
                     /label=repB
                     /note="required for primer synthesis"
                     /protein_id="ABM30613.1"
                     /translation="MKNDRTLQAIGRQLKAMGCERFDIGVRDATTGQMMNREWSAAEVL
                     QNTPWLKRMNAQGNDVYIRPAEQERHGLVLVDDLSEFDLDDMKAEGREPALVVETSPKN
                     YQAWVKVADAAGGELRGQIARTLASEYDADPASADSRHYGRLAGFTNRKDKHTTRAGYQ
                     PWVLLRESKGKTATAGPALVQQAGQQIEQAQRQQEKARRLASLELPERQLSRHRRTALD
                     EYRSEMAGLVKRFGDDLSKCDFIAAQKLASRGRSAEEIGKAMAEASPALAERKPGHEAD
                     YIERTVSKVMGLPSVQLARAELARAPAPRQRGMDRGGPDFSM"
     gene            6270..7241
                     /gene="repB"
                     /label=repB
     CDS             7302..7514
                     /codon_start=1
                     /gene="E"
                     /product="hypothetical protein"
                     /label=E
                     /note="unknown protein E"
                     /protein_id="ABM30614.1"
                     /translation="MEYEKSASGSVYLIKSDKGYWLPGGFGYTSNKAEAGRFSVADMAS
                     LNLDGCTLSLFREDKPFGPGKFLGD"
     gene            7302..7514
                     /gene="E"
                     /label=E
     CDS             7516..7722
                     /codon_start=1
                     /gene="F"
                     /product="repressor protein F"
                     /label=F
                     /note="regulates repAC operon"
                     /protein_id="ABM30615.1"
                     /translation="MKDQKDKQTGDLLASPDAVRQARYAERMKAKGMRQRKFWLTDDEY
                     EALRECLEELRAAQGGGSDPASA"
     gene            7516..7722
                     /gene="F"
                     /label=F
     misc_RNA        7685..7759
                     /product="regulatory RNA"
     gene            7739..8591
                     /gene="repA"
                     /label=repA
     regulatory      7739..7745
                     /regulatory_class="ribosome_binding_site"
                     /gene="repA"
     CDS             7752..8591
                     /codon_start=1
                     /gene="repA"
                     /product="replication protein A"
                     /label=repA
                     /note="hexameric replicative helicase; ssDNA-dependent 
                     ATPase"
                     /protein_id="ABM30616.1"
                     /translation="MATHKPINILEAFAAAPPPLDYVLPNMVAGTVGALVSPGGAGKSM
                     LALQLAAQIAGGPDLLEVGELPTGPVIYLPAEDPPTAIHHRLHALGAHLSAEERQAVAD
                     GLLIQPLIGSLPNIMAPEWFDGLKRAAEGRRLMVLDTLRRFHIEEENASGPMAQVIGRM
                     EAIAADTGCSIVFLHHASKGAAMMGAGDQQQASRGSSVLVDNIRWQSYLSSMTSAEAEE
                     WGVDDDQRRFFVRFGVSKANYGAPFADRWFRRHDGGVLKPAVLERQRKSKGVPRGEA"
     CDS             8578..9429
                     /codon_start=1
                     /gene="repC"
                     /product="replication protein C"
                     /label=repC
                     /note="required for origin recognition"
                     /protein_id="ABM30617.1"
                     /translation="MVKPKNKHSLSHVRHDPAHCLAPGLFRALKRGERKRSKLDVTYDY
                     GDGKRIEFSGPEPLGADDLRILQGLVAMAGPNGLVLGPEPKTEGGRQLRLFLEPKWEAV
                     TAECHVVKGSYRALAKEIGAEVDSGGALKHIQDCIERLWKVSIIAQNGRKRQGFRLLSE
                     YASDEADGRLYVALNPLIAQAVMGGGQHVRISMDEVRALDSETARLLHQRLCGWIDPGK
                     TGKASIDTLCGYVWPSEASGSTMRKRRQRVREALPELVALGWTVTEFAAGKYDITRPKA
                     AG"
     gene            8578..9429
                     /gene="repC"
                     /label=repC
     CDS             9737..10525
                     /codon_start=1
                     /gene="sulII"
                     /product="sulfonamid resistance protein"
                     /label=sulII
                     /protein_id="ABM30618.1"
                     /translation="MNKSLIIFGIVNITSDSFSDGGRYLAPDAAIAQARKLMAEGADVI
                     DLVRHPAIPTPRLFRPTQKSRVCAGAGRAQADGIPVSLDSYQPATQAYALSRGVAYLND
                     IRGFPDAAFYPQLAKSSAKLVVMHSVQDGQADRREAPAGDIMDHIAAFFDARIAALTGA
                     GIKRNRLVLDPGMGFFLGAAPETSLSVLARFDELRLRFDLPVLLSVSRKSFLRALTGRG
                     PGVSGPRHSLQSLPPPQVELTSSAHTSRAPCATGWRYWRR"
     gene            9737..10525
                     /gene="sulII"
                     /label=sulII