LOCUS       Exported                9388 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Promoter probe vector pPR9TT, complete sequence.
ACCESSION   AF187996
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9388)
  AUTHORS   Santos PM, Blatny JM, Di Bartolo I, Valla S, Zennaro E.
  TITLE     Physiological analysis of the expression of the styrene degradation 
            gene cluster in Pseudomonas fluorescens ST
  JOURNAL   Appl. Environ. Microbiol. 66 (4), 1305-1310 (2000)
  PUBMED    10742204
REFERENCE   2  (bases 1 to 9388)
  AUTHORS   Santos PM, Di Bartolo I, Blatny JM, Zennaro E, Valla S.
  TITLE     New broad-host-range promoter probe vectors based on the plasmid RK2
            replicon
  JOURNAL   FEMS Microbiol. Lett. 195 (1), 91-96 (2001)
  PUBMED    11167001
REFERENCE   3  (bases 1 to 9388)
  AUTHORS   Santos PM, Blatny JM, Di Bartolo I, Valla S, Zennaro E.
  TITLE     Direct Submission
  JOURNAL   Submitted (16-SEP-1999) Department of Biology, Third University of 
            Rome, Viale Marconi 446, Rome 00146, Italy
REFERENCE   4  (bases 1 to 9388)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..9388
                     /organism="Promoter probe vector pPR9TT"
                     /mol_type="other DNA"
                     /db_xref="taxon:110995"
     oriT            67..175
                     /note="incP origin of transfer"
     oriT            67..130
                     /label=origin of transfer from RK2
                     /note="origin of transfer from RK2"
     promoter        352..456
                     /gene="bla"
                     /label=AmpR promoter
     CDS             457..1317
                     /codon_start=1
                     /gene="bla"
                     /product="Bla"
                     /label=bla
                     /note="beta-lactamase; confers ampicillin resistance"
                     /protein_id="AAG41772.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            457..1317
                     /gene="bla"
                     /label=bla
     misc_feature    1563..2088
                     /note="origin of vegetative replication from RK2; oriV"
     rep_origin      1564..2094
                     /label=oriV
                     /note="incP origin of replication"
     regulatory      2316..2496
                     /regulatory_class="promoter"
                     /note="Pneo"
     CDS             2486..3634
                     /codon_start=1
                     /gene="trfA"
                     /product="TrfA"
                     /label=trfA
                     /note="initiator protein necessary for plasmid replication"
                     /protein_id="AAG41773.1"
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     MVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     gene            2486..3634
                     /gene="trfA"
                     /label=trfA
     primer_bind     complement(3815..3831)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             4225..4986
                     /codon_start=1
                     /gene="cat"
                     /product="Cat"
                     /label=cat
                     /note="chloramphenicol acetyltransferase; confers 
                     chloramphenicol resistance"
                     /protein_id="AAG41774.1"
                     /translation="MRHVRGSNFHHNEIRSLPGVFFELSRFSGAKEAKMEKKITGYTTV
                     DISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFLKTVKKNKHKFYPAFIHILARLMNAH
                     PELRMAMKDGELVIWDSVHPCYTVFHEQTETFSSLWSEYHDDFRQFLHIYSQDVACYGE
                     NLAYFPKGFIENMFFVSANPWVSFTSFDLNVANMDNFFAPVFTMGKYYTQGDKVLMPLA
                     IQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     gene            4225..4986
                     /gene="cat"
                     /label=cat
     CDS             4327..4986
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPELRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     regulatory      5150..5724
                     /regulatory_class="terminator"
                     /note="bidirectional"
     primer_bind     5775..5791
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(5810..8860)
                     /codon_start=1
                     /gene="lacZ"
                     /product="LacZ"
                     /label=lacZ
                     /note="beta-galactosidase; lacking first eight amino acids"
                     /protein_id="AAG41775.1"
                     /translation="PVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLR
                     SLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNVTYPITVNP
                     PFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVNSAFHLWCNGRWVGYGQDSRLPSE
                     FDLSAFLRAGENRLAVMVLRWSDGSYLEDQDMWRMSGIFRDVSLLHKPTTQISDFHVAT
                     RFNDDFSRAVLEAEVQMCGELRDYLRVTVSLWQGETQVASGTAPFGGEIIDERGGYADR
                     VTLRLNVENPKLWSAEIPNLYRAVVELHTADGTLIEAEACDVGFREVRIENGLLLLNGK
                     PLLIRGVNRHEHHPLHGQVMDEQTMVQDILLMKQNNFNAVRCSHYPNHPLWYTLCDRYG
                     LYVVDEANIETHGMVPMNRLTDDPRWLPAMSERVTRMVQRDRNHPSVIIWSLGNESGHG
                     ANHDALYRWIKSVDPSRPVQYEGGGADTTATDIICPMYARVDEDQPFPAVPKWSIKKWL
                     SLPGETRPLILCEYAHAMGNSLGGFAKYWQAFRQYPRLQGGFVWDWVDQSLIKYDENGN
                     PWSAYGGDFGDTPNDRQFCMNGLVFADRTPHPALTEAKHQQQFFQFRLSGQTIEVTSEY
                     LFRHSDNELLHWMVALDGKPLASGEVPLDVAPQGKQLIELPELPQPESAGQLWLTVRVV
                     QPNATAWSEAGHISAWQQWRLAENLSVTLPAASHAIPHLTTSEMDFCIELGNKRWQFNR
                     QSGFLSQMWIGDKKQLLTPLRDQFTRAPLDNDIGVSEATRIDPNAWVERWKAAGHYQAE
                     AALLQCTADTLADAVLITTAHAWQHQGKTLFISRKTYRIDGSGQMAITVDVEVASDTPH
                     PARIGLNCQLAQVAERVNWLGLGPQENYPDRLTAACFDRWDLPLSDMYTPYVFPSENGL
                     RCGTRELNYGPHQWRGDFQFNISRYSQQQLMETSHRHLLHAEEGTWLNIDGFHMGIGGD
                     DSWSPSVSAEFQLSAGRYHYQLVWCQK"
     gene            complement(5810..8860)
                     /gene="lacZ"
                     /label=lacZ
     CDS             complement(5810..8857)
                     /codon_start=1
                     /gene="lacZ"
                     /product="beta-galactosidase"
                     /label=lacZ
                     /translation="VVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRS
                     LNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNVTYPITVNPP
                     FVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVNSAFHLWCNGRWVGYGQDSRLPSEF
                     DLSAFLRAGENRLAVMVLRWSDGSYLEDQDMWRMSGIFRDVSLLHKPTTQISDFHVATR
                     FNDDFSRAVLEAEVQMCGELRDYLRVTVSLWQGETQVASGTAPFGGEIIDERGGYADRV
                     TLRLNVENPKLWSAEIPNLYRAVVELHTADGTLIEAEACDVGFREVRIENGLLLLNGKP
                     LLIRGVNRHEHHPLHGQVMDEQTMVQDILLMKQNNFNAVRCSHYPNHPLWYTLCDRYGL
                     YVVDEANIETHGMVPMNRLTDDPRWLPAMSERVTRMVQRDRNHPSVIIWSLGNESGHGA
                     NHDALYRWIKSVDPSRPVQYEGGGADTTATDIICPMYARVDEDQPFPAVPKWSIKKWLS
                     LPGETRPLILCEYAHAMGNSLGGFAKYWQAFRQYPRLQGGFVWDWVDQSLIKYDENGNP
                     WSAYGGDFGDTPNDRQFCMNGLVFADRTPHPALTEAKHQQQFFQFRLSGQTIEVTSEYL
                     FRHSDNELLHWMVALDGKPLASGEVPLDVAPQGKQLIELPELPQPESAGQLWLTVRVVQ
                     PNATAWSEAGHISAWQQWRLAENLSVTLPAASHAIPHLTTSEMDFCIELGNKRWQFNRQ
                     SGFLSQMWIGDKKQLLTPLRDQFTRAPLDNDIGVSEATRIDPNAWVERWKAAGHYQAEA
                     ALLQCTADTLADAVLITTAHAWQHQGKTLFISRKTYRIDGSGQMAITVDVEVASDTPHP
                     ARIGLNCQLAQVAERVNWLGLGPQENYPDRLTAACFDRWDLPLSDMYTPYVFPSENGLR
                     CGTRELNYGPHQWRGDFQFNISRYSQQQLMETSHRHLLHAEEGTWLNIDGFHMGIGGDD
                     SWSPSVSAEFQLSAGRYHYQLVWCQK"
     misc_feature    8860..8931
                     /label=multicloning site
                     /note="multicloning site"
     primer_bind     complement(8897..8913)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      complement(8961..9365)
                     /regulatory_class="terminator"
                     /note="rrnBT1T2 terminators"
     terminator      8964..8991
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      9083..9169
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"