LOCUS       Exported                6810 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Promoter probe vector pPROBE-NT, complete sequence.
ACCESSION   AF286454
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6810)
  AUTHORS   Miller WG, Leveau JH, Lindow SE.
  TITLE     Improved gfp and inaZ broad-host-range promoter-probe vectors
  JOURNAL   Mol. Plant Microbe Interact. 13 (11), 1243-1250 (2000)
  PUBMED    11059491
REFERENCE   2  (bases 1 to 6810)
  AUTHORS   Leveau JHJ., Miller WG, Lindow SE.
  TITLE     Direct Submission
  JOURNAL   Submitted (13-JUL-2000) Plant & Microbial Biology, University of 
            California, 111 Koshland Hall, Berkeley, CA 94720-3102, USA
REFERENCE   3  (bases 1 to 6810)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6810
                     /organism="Promoter probe vector pPROBE-NT"
                     /mol_type="genomic DNA"
                     /label=derived from plasmid pBBR1MCS-2
                     /note="derived from plasmid pBBR1MCS-2"
                     /db_xref="taxon:140683"
     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
                     /gene="rep"
                     /product="Rep"
                     /label=rep
                     /note="required for plasmid replication"
                     /protein_id="AAG29459.1"
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
     gene            1793..2455
                     /gene="rep"
                     /label=rep
     primer_bind     3162..3178
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        3188..3206
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      3239..3411
                     /regulatory_class="terminator"
                     /note="T1 rrnB terminator region"
     terminator      3247..3333
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     regulatory      3420..3592
                     /regulatory_class="terminator"
                     /note="T1 rrnB terminator region"
     terminator      3428..3514
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     regulatory      3601..3773
                     /regulatory_class="terminator"
                     /note="T1 rrnB terminator region"
     terminator      3609..3695
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     regulatory      3782..3954
                     /regulatory_class="terminator"
                     /note="T1 rrnB terminator region"
     terminator      3790..3876
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     misc_feature    3973..4022
                     /note="multicloning site; EcoRI to HindIII"
     regulatory      4055..4060
                     /regulatory_class="ribosome_binding_site"
     CDS             4069..4785
                     /codon_start=1
                     /gene="gfp"
                     /product="GFP"
                     /label=gfp
                     /note="green fluorescent protein; contains F64L and S65T 
                     mutations"
                     /protein_id="AAG29458.1"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     gene            4069..4785
                     /gene="gfp"
                     /label=gfp
     regulatory      4795..4967
                     /regulatory_class="terminator"
                     /note="T1 rrnB terminator region"
     terminator      4803..4889
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     misc_feature    4984..3219
                     /label=pBBR1MCS-2 backbone
                     /note="pBBR1MCS-2 backbone"
     promoter        complement(5001..5019)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5040..5056)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5064..5080
                     /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)."
     promoter        complement(5088..5118)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5133..5154
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             complement(5559..6353)
                     /codon_start=1
                     /gene="npt"
                     /product="kanamycin phosphotransferase"
                     /label=npt
                     /note="kanamycin resistance determinant"
                     /protein_id="AAG29460.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     gene            complement(5559..6353)
                     /gene="npt"
                     /label=npt
     CDS             complement(6611..799)
                     /codon_start=1
                     /product="Mob"
                     /label=Mob
                     /note="required for plasmid mobilization"
                     /protein_id="AAG29461.1"
                     /translation="MAAYAIMRCKKLAKMGNVAASLKHAYRERETPNADASRTPENEHW
                     AASSTDEAMGRLRELLPEKRRKDAVLAVEYVMTASPEWWKSASQEQQAAFFEKAHKWLA
                     DKYGADRIVTASIHRDETSPHMTAFVVPLTQDGRLSAKEFIGNKAQMTRDQTTFAAAVA
                     DLGLQRGIEGSKARHTRIQAFYEALERPPVGHVTISPQAVEPRAYAPQGLAEKLGISKR
                     VETPEAVADRLTKAVRQGYEPALQAAAGAREMRKKADQAQETARDLRERLKPVLDALGP
                     LNRDMQAKAAAIIKAVGEKLLTEQREVQRQKQAQRQQERGRAHFPEKCHLG"