LOCUS       Exported                6759 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pPM2a, complete sequence.
ACCESSION   GQ139362
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6759)
  AUTHORS   Martins PM, Lau IF, Bacci M, Belasque J, do Amaral AM, Taboga SR, 
            Ferreira H.
  TITLE     Subcellular localization of proteins labeled with GFP in Xanthomonas
            citri ssp. citri: targeting the division septum
  JOURNAL   FEMS Microbiol. Lett. 310 (1), 76-83 (2010)
  PUBMED    20629754
REFERENCE   2  (bases 1 to 6759)
  AUTHORS   Martins PMM., Ferreira H, Lau I.
  TITLE     An expression vector for GFP-tagging of proteins in Xanthomonas 
            axonopodis pv. citri
  JOURNAL   Unpublished
REFERENCE   3  (bases 1 to 6759)
  AUTHORS   Martins PMM., Ferreira H, Lau I.
  TITLE     Direct Submission
  JOURNAL   Submitted (07-MAY-2009) UNESP, Rua Cristovao Colombo, 2265, Sao Jose
            do Rio Preto, SP 15054000, Brazil
REFERENCE   4  (bases 1 to 6759)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6759
                     /organism="Expression vector pPM2a"
                     /mol_type="other DNA"
                     /note="expression vector for GFP-tagged protein expression 
                     in Xanthomonas axonopodis pv. citri"
                     /db_xref="taxon:651132"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        143..173
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /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     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(1050..2102)
                     /codon_start=1
                     /gene="xylR"
                     /product="pxyl promoter repressor"
                     /label=xylR
                     /protein_id="ACS28252.1"
                     /translation="MTGLNKSTVSSQVNTLMKESMVFEIGQGQSSGGRRPVMLVFNKKA
                     GYSVGIDVGVDYINGILTDLEGTIVLDQYRHLESNSPEITKDILIDMIHHFITQMPQSP
                     YGFIGIGICVPGLIDKDQKIVFTPNSNWRDIDLKSSIQEKYNVSVFIENEANAGAYGEK
                     LFGAAKNHDNIIYVSISTGIGIGVIINNHLYRGVSGFSGEMGHMTIDFNGPKCSCGNRG
                     CWELYASEKALLKSLQTKEKKLSYQDIINLAHLNDIGTLNALQNFGFYLGIGLTNILNT
                     FNPQAVILRNSIIESHPMVLNSMRSEVSSRVYSQLGNSYELLPSSLGQNAPALGMSSIV
                     IDHFLDMITM"
     gene            complement(1050..2102)
                     /gene="xylR"
                     /label=xylR
     CDS             2456..3202
                     /codon_start=1
                     /gene="gfp"
                     /product="green fluorescent protein"
                     /label=gfp
                     /protein_id="ACS28253.1"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFAYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYKEAGGRALNLE"
     gene            2456..3202
                     /gene="gfp"
                     /label=gfp
     CDS             2456..3169
                     /codon_start=1
                     /product="Aequoria victoria green fluorescent protein"
                     /label=GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFAYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(3218..3234)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      3375..3803
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     CDS             4147..4941
                     /codon_start=1
                     /gene="aph"
                     /product="aminoglicoside phosphotransferase"
                     /label=aph
                     /protein_id="ACS28254.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     gene            4147..4941
                     /gene="aph"
                     /label=aph
     CDS             4959..5819
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=bla
                     /protein_id="ACS28255.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            4959..5819
                     /gene="bla"
                     /label=bla
     rep_origin      5990..6578
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"