LOCUS       Exported                7382 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Promoter probe vector pPROBE-gfp[LVA], complete sequence.
ACCESSION   AF286461
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7382)
  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 7382)
  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 7382)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7382
                     /organism="Promoter probe vector pPROBE-gfp[LVA]"
                     /mol_type="genomic DNA"
                     /label=derived from plasmid pBBR1MCS-2
                     /note="derived from plasmid pBBR1MCS-2"
                     /db_xref="taxon:140686"
     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="AAG29487.1"
                     /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    3968..4024
                     /note="multicloning site; HindIII to EcoRI"
     regulatory      4052..4057
                     /regulatory_class="ribosome_binding_site"
     CDS             4066..4821
                     /codon_start=1
                     /gene="gfp[LVA]"
                     /product="GFP[LVA]"
                     /label=gfp[LVA]
                     /note="unstable green fluorescent protein; contains F64L 
                     and S65T mutations and C-terminal tag"
                     /protein_id="AAG29486.1"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYKRPAANDENYALVA"
     gene            4066..4821
                     /gene="gfp[LVA]"
                     /label=gfp[LVA]
     CDS             4066..4779
                     /codon_start=1
                     /product="Aequoria victoria green fluorescent protein"
                     /label=GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     regulatory      4844..4938
                     /regulatory_class="terminator"
                     /note="lambda t0 terminator region"
     terminator      4844..4938
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             4982..5641
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     regulatory      5700..5798
                     /regulatory_class="terminator"
                     /note="T1 rrnB terminator region"
     terminator      5706..5792
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     misc_feature    5833..3219
                     /label=pBBR1MCS-2 backbone
                     /note="pBBR1MCS-2 backbone"
     CDS             complement(6131..6925)
                     /codon_start=1
                     /gene="npt"
                     /product="kanamycin phosphotransferase"
                     /label=npt
                     /note="kanamycin resistance determinant"
                     /protein_id="AAG29488.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     gene            complement(6131..6925)
                     /gene="npt"
                     /label=npt
     CDS             complement(7183..799)
                     /codon_start=1
                     /product="Mob"
                     /label=Mob
                     /note="required for plasmid mobilization"
                     /protein_id="AAG29489.1"
                     /translation="MAAYAIMRCKKLAKMGNVAASLKHAYRERETPNADASRTPENEHW
                     AASSTDEAMGRLRELLPEKRRKDAVLAVEYVMTASPEWWKSASQEQQAAFFEKAHKWLA
                     DKYGADRIVTASIHRDETSPHMTAFVVPLTQDGRLSAKEFIGNKAQMTRDQTTFAAAVA
                     DLGLQRGIEGSKARHTRIQAFYEALERPPVGHVTISPQAVEPRAYAPQGLAEKLGISKR
                     VETPEAVADRLTKAVRQGYEPALQAAAGAREMRKKADQAQETARDLRERLKPVLDALGP
                     LNRDMQAKAAAIIKAVGEKLLTEQREVQRQKQAQRQQERGRAHFPEKCHLG"