LOCUS       Exported                7682 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Promoter probe vector pJB785TTKm1, complete sequence.
ACCESSION   AF320510
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7682)
  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   2  (bases 1 to 7682)
  AUTHORS   Santos PM, Di Bartolo I, Blatny JM, Zennaro E, Valla S.
  TITLE     Direct Submission
  JOURNAL   Submitted (09-NOV-2000) Biology, University of Rome Roma Tre, 
            Viale Marconi 446, Rome, RM 00146, Italy
REFERENCE   3  (bases 1 to 7682)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7682
                     /organism="Promoter probe vector pJB785TTKm1"
                     /mol_type="other DNA"
                     /db_xref="taxon:152797"
     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="AAK13423.1"
                     /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="AAK13424.1"
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     MVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     gene            2486..3634
                     /gene="trfA"
                     /label=trfA
     CDS             4248..4946
                     /codon_start=1
                     /gene="kan"
                     /product="aminoglycoside-3-phosphotransferase"
                     /label=kan
                     /note="confers kanamycin resistance"
                     /protein_id="AAK13425.1"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAAC"
     gene            4248..4946
                     /gene="kan"
                     /label=kan
     regulatory      5010..5476
                     /regulatory_class="terminator"
                     /note="bidirectional"
     CDS             complement(5588..7267)
                     /codon_start=1
                     /gene="luc"
                     /product="luciferase"
                     /label=luc
                     /note="fluorescent protein"
                     /protein_id="AAK13426.1"
                     /translation="MAFLRRSAPMEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALV
                     PGTIAFTDAHIEVNITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGA
                     LFIGVAVAPANDIYNERELLNSMNISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDS
                     KTDYQGFQSMYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHR
                     TACVRFSHARDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEEL
                     FLRSLQDYKIQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRF
                     HLPGIRQGYGLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGEL
                     CVRGPMIMSGYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQV
                     APAELESILLQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTT
                     AKKLRGGVVFVDEVPKGLTGKLDARKIREILIKAKKGGKSKL"
     gene            complement(5588..7267)
                     /gene="luc"
                     /label=luc
     CDS             complement(5588..7240)
                     /codon_start=1
                     /gene="luc"
                     /product="firefly luciferase"
                     /label=luciferase
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVNITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMNISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKSKL"
     terminator      7258..7285
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     misc_feature    complement(7267..7300)
                     /label=contains NcoI and BglII cloning sites
                     /note="contains NcoI and BglII cloning sites"
     regulatory      complement(7300..7612)
                     /regulatory_class="terminator"
                     /note="rrnBT1T2"
     terminator      7377..7463
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"