LOCUS       Exported               10690 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Broad host range expression vector pRK415, complete sequence.
ACCESSION   EF437940
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10690)
  AUTHORS   Hulter N, Wackernagel W.
  TITLE     Double illegitimate recombination events integrate DNA segments 
            through two different mechanisms during natural transformation of 
            Acinetobacter baylyi
  JOURNAL   Mol. Microbiol. 67 (5), 984-995 (2008)
  PUBMED    18194157
REFERENCE   2  (bases 1 to 10690)
  AUTHORS   Huelter N, Wackernagel W.
  TITLE     Direct Submission
  JOURNAL   Submitted (13-FEB-2007) Genetics, Department of Biology and 
            Environmental Sciences, Carl von Ossietzky University Oldenburg, 
            Carl-von-Ossietzkystrasse 9-11, Oldenburg 26111, Germany
REFERENCE   3  (bases 1 to 10690)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..10690
                     /organism="Broad host range expression vector pRK415"
                     /mol_type="other DNA"
                     /note="derived from plasmid pRK404 GenBank Accession Number
                     AY204475"
                     /db_xref="taxon:429723"
     rep_origin      1..712
                     /label=oriV
                     /note="incP origin of replication"
     rep_origin      276..987
                     /note="oriV; IncP"
     misc_feature    complement(707..860)
                     /label=similar to NC_001371 Plasmid ColE1
                     /note="similar to NC_001371 Plasmid ColE1"
     misc_feature    complement(852..1082)
                     /label=similar to NC_001371 Plasmid ColE1
                     /note="similar to NC_001371 Plasmid ColE1"
     CDS             complement(971..1312)
                     /codon_start=1
                     /gene="lacZ-alpha"
                     /product="beta-galactosidase alpha peptide"
                     /label=lacZ-alpha
                     /protein_id="ABO32178.1"
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRHASRREKGGQVSGKRQGRNRRAHEGA
                     SRGKRLVSL"
     gene            complement(971..1312)
                     /gene="lacZ-alpha"
                     /label=lacZ-alpha
     primer_bind     1222..1238
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    1239..1295
                     /label=multiple cloning site
                     /note="multiple cloning site"
     primer_bind     complement(1308..1324)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1332..1348
                     /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)."
     regulatory      1352..1421
                     /regulatory_class="promoter"
                     /note="lac promoter"
     promoter        complement(1356..1386)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1401..1422
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     gene            complement(1654..2279)
                     /gene="traJ"
                     /label=traJ
     CDS             complement(1654..2025)
                     /codon_start=1
                     /gene="traJ"
                     /product="TraJ"
                     /label=traJ
                     /note="oriT-recognizing protein"
                     /protein_id="ABO32179.1"
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            2058..2167
                     /label=incP
                     /note="incP"
     misc_feature    2299..2532
                     /label=similar to NC_001371 Plasmid ColE1
                     /note="similar to NC_001371 Plasmid ColE1"
     misc_feature    2524..2676
                     /label=similar to NC_001371 Plasmid ColE1
                     /note="similar to NC_001371 Plasmid ColE1"
     CDS             complement(2848..3498)
                     /codon_start=1
                     /gene="tetR"
                     /product="TetR"
                     /label=tetR
                     /note="tetracycline resistance regulation"
                     /protein_id="ABO32180.1"
                     /translation="MTKLQPNTVIRAALDLLNEVGVDGLTTRKLAERLGVQQPALYWHF
                     RNKRALLDALAEAMLAENHTHSVPRADDDWRSFLIGNARSFRQALLAYRDGARIHAGTR
                     PGAPQMETADAQLRFLCEAGFSAGDAVNALMTISYFTVGAVLEEQAGDSDAGERGGTVE
                     QAPLSPLLRAAIDAFDEAGPDAAFEQGLAVIVDGLAKRRLVVRNVEGPRKGDD"
     gene            complement(2848..3498)
                     /gene="tetR"
                     /label=tetR
     CDS             3604..4803
                     /codon_start=1
                     /gene="tetA"
                     /product="tetracycline resistance protein"
                     /label=tetA
                     /protein_id="ABO32181.1"
                     /translation="MKPNIPLIVILSTVALDAVGIGLIMPVLPGLLRDLVHSNDVTAHY
                     GILLALYALVQFACAPVLGALSDRFGRRPILLVSLAGATVDYAIMATAPFLWVLYIGRI
                     VAGITGATGAVAGAYIADITDGDERARHFGFMSACFGFGMVAGPVLGGLMGGFSPHAPF
                     FAAAALNGLNFLTGCFLLPESHKGERRPLRREALNPLASFRWARGMTVVAALMAVFFIM
                     QLVGQVPAALWVIFGEDRFHWDATTIGISLAAFGILHSLAQAMITGPVAARLGERRALM
                     LGMIADGTGYILLAFATRGWMAFPIMVLLASGGIGMPALQAMLSRQVDEERQGQLQGSL
                     AALTSLTSIVGPLLFTAIYAASITTWNGWAWIAGAALYLLCLPALRRGLWSGAGQRADR
                     "
     gene            3604..4803
                     /gene="tetA"
                     /label=tetA
     CDS             complement(5434..6099)
                     /codon_start=1
                     /gene="upf16.5"
                     /product="unknown plasmid factor 16.5"
                     /label=upf16.5
                     /protein_id="ABO32182.1"
                     /translation="MTWCIANARALWGQFRLGVQQPALYWHFRNKRALLDALASLSIAR
                     AHGALYELPINRGLKLTIVIKAQIRRLGAADVQDFREIRLSALKKAPEMFGSVYEHEEK
                     KPMEAFAERLRDAVAFGAYIDGEIIGLSVFKQEDGPKDAHKAHLSGVFVEPEQRGRGVA
                     GMLLRALPAGLLLVMIVRQIPTGIWWMRIFILGALNISLFWSLLFISVYRLPGGSRRR"
     gene            complement(5434..6099)
                     /gene="upf16.5"
                     /label=upf16.5
     CDS             complement(6074..7222)
                     /codon_start=1
                     /gene="trfA2"
                     /product="trans-acting replication A2"
                     /label=trfA2
                     /protein_id="ABO32183.1"
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     MVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     gene            complement(6074..7222)
                     /gene="trfA2"
                     /label=trfA2
     CDS             complement(6074..6931)
                     /codon_start=1
                     /gene="trfA1"
                     /product="trans-acting replication A1"
                     /label=trfA1
                     /protein_id="ABO32184.1"
                     /translation="MTTKKRKTAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAP
                     WPDTMRAMPNDTARSALFTTRNKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELV
                     WQQVLEYAKRTPIGEPITFTFYELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRV
                     GHLESVSLLHRFRVLDRGKKTSRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRM
                     FDYFSSHREPYPLKLETFRLMCGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLV
                     HCKR"
     gene            complement(6074..6931)
                     /gene="trfA1"
                     /label=trfA1
     CDS             complement(7271..7621)
                     /codon_start=1
                     /gene="ssb"
                     /product="single-stranded DNA binding protein"
                     /label=ssb
                     /protein_id="ABO32185.1"
                     /translation="MSHNQFQFIGNLTRDTEVRHGNSNKPQAIFDIAVNEEWRNDAGDK
                     QERTDFFRIKCFGSQAEAHGKYLGKGSLVFVQGKIRNTKYEKDGQTVYGTDFIADKVDY
                     LDTKAPGGSNQE"
     gene            complement(7271..7621)
                     /gene="ssb"
                     /label=ssb
     CDS             7742..8107
                     /codon_start=1
                     /gene="trbA"
                     /product="TrbA"
                     /label=trbA
                     /note="regulation of tra gene expression"
                     /protein_id="ABO32186.1"
                     /translation="MYNQIFFTNILRLLDERGMTKHELSERAGVSISFLSDLTNGKANP
                     SLKVMEAIADALETPLPLLLESTDLDREALAEIAGHPFKSSVPPGYERISVVLPSHKAF
                     IVKKWGDDTRKKLRGRL"
     gene            7742..8107
                     /gene="trbA"
                     /label=trbA
     CDS             complement(9145..10239)
                     /codon_start=1
                     /gene="incC1"
                     /product="IncC1"
                     /label=incC1
                     /note="IncC partitioning protein, long form"
                     /protein_id="ABO32187.1"
                     /translation="MGVIHEETAYRKPVPGGDPGAGSGAADHRDSAGRLSRWEATGDVR
                     NVAGTDQGRSVASGASRVGRVRGQELARGVRAGNGGSAGTSGVHRPEVGSGRQEKTGNQ
                     TMKTLVTANQKGGVGKTSTLVHLAFDFFERGLRVAVIDLDPQGNASYTLKDFATGLHAS
                     KLFGAVPAGGWTETAPAAGDGQAARLALIESNPVLANAERLSLDDARELFGANIKALAN
                     QGFDVCLIDTAPTLGVGLAAALFAADYVLSPIELEAYSIQGIKKMVTTIANVRQKNAKL
                     QFLGMVPSKVDARNPRHARHQAELLAAYPKMMIPATVGLRSSIADALASGVPVWKIKKT
                     AARKASKEVRALADYVFTKMEISQ"
     gene            complement(9145..10239)
                     /gene="incC1"
                     /label=incC1
     CDS             complement(9145..9924)
                     /codon_start=1
                     /gene="incC2"
                     /product="IncC2"
                     /label=incC2
                     /note="IncC partitioning protein, short form"
                     /protein_id="ABO32188.1"
                     /translation="MKTLVTANQKGGVGKTSTLVHLAFDFFERGLRVAVIDLDPQGNAS
                     YTLKDFATGLHASKLFGAVPAGGWTETAPAAGDGQAARLALIESNPVLANAERLSLDDA
                     RELFGANIKALANQGFDVCLIDTAPTLGVGLAAALFAADYVLSPIELEAYSIQGIKKMV
                     TTIANVRQKNAKLQFLGMVPSKVDARNPRHARHQAELLAAYPKMMIPATVGLRSSIADA
                     LASGVPVWKIKKTAARKASKEVRALADYVFTKMEISQ"
     gene            complement(9145..9924)
                     /gene="incC2"
                     /label=incC2
     CDS             complement(9921..10226)
                     /codon_start=1
                     /gene="korA"
                     /product="KorA"
                     /label=korA
                     /note="regulation of gene expression"
                     /protein_id="ABO32189.1"
                     /translation="MKKRLTESQFQEAIQGLEVGQQTIEIARGVLVDGKPQATFATSLG
                     LTRGAVSQAVHRVWAAFEDKNLPEGYARVTAVLPEHQAYIVRKWEADAKKKQETKR"
     gene            complement(9921..10226)
                     /gene="korA"
                     /label=korA