LOCUS       Exported                8530 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pHRS-9, complete sequence.
ACCESSION   AY289598
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8530)
  AUTHORS   Ramos HJO., Soares-Ramos JRL., Souza EM, Pedrosa FO.
  TITLE     Direct Submission
  JOURNAL   Submitted (01-MAY-2003) Department of Biochemistry and Molecular 
            Biology, Universidade Federal do Parana - UFPR, Centro Politecnico, 
            Curitiba, PR 81531-990, Brazil
REFERENCE   2  (bases 1 to 8530)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8530
                     /organism="Cloning vector pHRS-9"
                     /mol_type="genomic DNA"
                     /label=suicide vector for site-directed mutagenesis
                     /note="suicide vector for site-directed mutagenesis"
                     /db_xref="taxon:230832"
     misc_feature    1..28
                     /label=derived from suicide plasmid pSUP202
                     /note="derived from suicide plasmid pSUP202"
     misc_feature    29..838
                     /note="RS-alpha repetitive sequence from Bradyrhizobium 
                     japonicum CPAC7"
     misc_feature    338..412
                     /label=SstI-BamHI-BglII multiple cloning site
                     /note="SstI-BamHI-BglII multiple cloning site"
     gene            839..3334
                     /gene="nonfunctional tetA"
                     /label=nonfunctional tetA
                     /note="tetA was knocked out by cloning of the RS-alpha 
                     repetitive sequence into HindIII site of the pSUP202 
                     vector"
     CDS             896..2086
                     /codon_start=1
                     /gene="tet"
                     /product="tetracycline efflux protein"
                     /label=TcR
                     /note="confers resistance to tetracycline"
                     /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
                     GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
                     VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
                     LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
                     QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
                     AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
                     AALTSLTSITGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
     CDS             2727..2918
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     misc_feature    3020..3160
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      3334..3944
                     /label=pBR322
                     /note="pBR322"
     rep_origin      complement(3346..3934)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4105..4965)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=bla
                     /protein_id="AAP42291.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(4105..4965)
                     /gene="bla"
                     /label=bla
     promoter        complement(4966..5070)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5273..5375
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             5376..6035
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=cat
                     /protein_id="AAP42292.1"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     gene            5376..6035
                     /gene="cat"
                     /label=cat
     misc_feature    6294..8021
                     /label=mob site from RP4
                     /note="mob site from RP4"
     oriT            6916..7025
                     /note="incP origin of transfer"
     CDS             7058..7429
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"