LOCUS       Exported                9233 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pHRGFPTC, complete sequence.
ACCESSION   AY237646
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9233)
  AUTHORS   Quandt J, Hynes MF.
  TITLE     Versatile suicide vectors which allow direct selection for gene 
            replacement in gram-negative bacteria
  JOURNAL   Gene 127 (1), 15-21 (1993)
  PUBMED    8486283
REFERENCE   2  (bases 1 to 9233)
  AUTHORS   Dennis JJ, Zylstra GJ.
  TITLE     Plasposons: modular self-cloning minitransposon derivatives for 
            rapid genetic analysis of gram-negative bacterial genomes
  JOURNAL   Appl. Environ. Microbiol. 64 (7), 2710-2715 (1998)
  PUBMED    9647854
REFERENCE   3  (bases 1 to 9233)
  AUTHORS   Ouahrani-Bettache S, Porte F, Teyssier J, Liautard JP, Kohler S.
  TITLE     pBBR1-GFP: a broad-host-range vector for prokaryotic promoter 
            studies
  JOURNAL   BioTechniques 26 (4), 620-622 (1999)
  PUBMED    10343896
REFERENCE   4  (bases 1 to 9233)
  AUTHORS   Ramos HJ, Roncato-Maccari LD, Souza EM, Soares-Ramos JR, Hungria M, 
            Pedrosa FO.
  TITLE     Monitoring Azospirillum-wheat interactions using the gfp and gusA 
            genes constitutively expressed from a new broad-host range vector
  JOURNAL   J. Biotechnol. 97 (3), 243-252 (2002)
  PUBMED    12084480
REFERENCE   5  (bases 1 to 9233)
  AUTHORS   Ramos HJO., Soares-Ramos JRL., Souza EM, Pedrosa FO.
  TITLE     Direct Submission
  JOURNAL   Submitted (14-FEB-2003) Department of Biochemistry and Molecular 
            Biology, Universidade Federal do Parana - UFPR, Centro Politecnico, 
            Curitiba, PR 81531-990, Brazil
REFERENCE   6  (bases 1 to 9233)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..9233
                     /organism="Cloning vector pHRGFPTC"
                     /mol_type="genomic DNA"
                     /label=broad-host-range vector expressing GFP
                     /note="broad-host-range vector expressing GFP"
                     /db_xref="taxon:223837"
     misc_feature    1..9233
                     /note="Mobilization (mob) and replication (rep) regions are
                     derived from the broad-host-range vector pBBR1 isolated 
                     from Bordetella bronchiseptica. The expression of cat and 
                     gfp genes is under control of gentamycin resistance aacC1 
                     promoter. The pgen promoter and part of the gentamycin gene
                     (aacC1) were isolated from pJQ200KS as a BglII/BamHI 
                     fragment."
     misc_feature    1..2642
                     /note="mob and rep regions from broad-host-range vector 
                     pBBR1-GFP as deposited in GenBank Accession number 
                     AF110459"
     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
                     /product="replication protein for the broad-host-range 
                     plasmid pBBR1 from Bordetella bronchiseptica"
                     /label=pBBR1 Rep
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
     primer_bind     2614..2630
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2664..2680)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(3110..3130)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     intron          3260..3325
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             complement(3491..4150)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol-acetyl-transferase"
                     /label=cat
                     /note="CAT"
                     /protein_id="AAO63170.1"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     gene            complement(3491..4150)
                     /gene="cat"
                     /label=cat
                     /note="promoterless cat gene from pBBR1-GFP as deposited in
                     GenBank Accession number AF110459"
     CDS             complement(4213..4929)
                     /codon_start=1
                     /gene="gfp"
                     /product="green fluorescent protein"
                     /label=gfp
                     /note="promoterless gfp gene from pBBR1-GFP; modified form 
                     mut3 from Aequorea victoria GFP as deposited in GenBank 
                     Accession number M62653"
                     /protein_id="AAO63171.1"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     gene            complement(4213..4929)
                     /gene="gfp"
                     /label=gfp
                     /note="promoterless gfp gene from pBBR1-GFP; modified form 
                     mut3 from pBBR1-GFP"
     misc_feature    4966..5465
                     /label=from pJQ200KS
                     /note="from pJQ200KS"
     regulatory      complement(5472..5501)
                     /regulatory_class="promoter"
                     /note="-35 and -10 promoter regions of the gentamycin 
                     resistance aacC1 gene from pJQ200KS"
     promoter        complement(5473..5501)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     misc_feature    5510..6178
                     /label=from pJQ200KS
                     /note="from pJQ200KS"
     protein_bind    6010..6031
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6046..6076
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6084..6100
                     /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)."
     primer_bind     6108..6124
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        6145..6163
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        6192..6221
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     regulatory      6197..6232
                     /regulatory_class="promoter"
                     /note="tetA promoter"
     CDS             6269..7459
                     /codon_start=1
                     /gene="tetA"
                     /product="tetracyclin resistance protein class C"
                     /label=tetA
                     /note="tetA gene from p34S-Tc as deposited in GenBank 
                     Accession number AF62082"
                     /protein_id="AAO63172.1"
                     /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
                     GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
                     VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
                     LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
                     QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
                     AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
                     AALTSLTSITGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
     gene            6269..7459
                     /gene="tetA"
                     /label=tetA
     CDS             complement(8162..9022)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=bla
                     /protein_id="AAO63173.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(8162..9022)
                     /gene="bla"
                     /label=bla
                     /note="bla gene from pBBR1-GFP"