LOCUS       Exported                6651 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pHF5Na, complete sequence.
ACCESSION   FJ573043
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6651)
  AUTHORS   Martins PMM., Oehlmeyer T, Amaral AM, Ferreira H.
  TITLE     A protein expression system for Tandem Affinity purification in 
            Xanthomonas axonopodis pv. citri
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6651)
  AUTHORS   Martins PMM., Oehlmeyer T, Amaral AM, Ferreira H.
  TITLE     Direct Submission
  JOURNAL   Submitted (18-DEC-2008) Biologia, UNESP, Rua Cristovao Colombo, 
            2265, 15054000, SP, Brazil
REFERENCE   3  (bases 1 to 6651)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6651
                     /organism="Expression vector pHF5Na"
                     /mol_type="other DNA"
                     /db_xref="taxon:590084"
     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(237..1289)
                     /codon_start=1
                     /gene="xylR"
                     /product="XylR"
                     /label=xylR
                     /note="xylose repressor from xylose operon"
                     /protein_id="ACM16581.1"
                     /translation="MTGLNKSTVSSQVNTLMKESMVFEIGQGQSSGGRRPVMLVFNKKA
                     GYSVGIDVGVDYINGILTDLEGTIVLDQYRHLESNSPEITKDILIDMIHHFITQMPQSP
                     YGFIGIGICVPGLIDKDQKIVFTPNSNWRDIDLKSSIQEKYNVSVFIENEANAGAYGEK
                     LFGAAKNHDNIIYVSISTGIGIGVIINNHLYRGVSGFSGEMGHMTIDFNGPKCSCGNRG
                     CWELYASEKALLKSLQTKEKKLSYQDIINLAHLNDIGTLNALQNFGFYLGIGLTNILNT
                     FNPQAVILRNSIIESHPMVLNSMRSEVSSRVYSQLGNSYELLPSSLGQNAPALGMSSIV
                     IDHFLDMITM"
     gene            complement(237..1289)
                     /gene="xylR"
                     /label=xylR
     CDS             1643..2269
                     /codon_start=1
                     /gene="tap"
                     /product="protein A-TEV-CBP"
                     /label=tap
                     /protein_id="ACM16582.1"
                     /translation="MITSYSIHYTKLSMAGLAQHDEAVDNKFNKEQQNAFYEILHLPNL
                     NEEQRNAFIQSLKDDPSQSANLLAEAKKLNDAQAPKVDNKFNKEQQNAFYEILHLPNLN
                     EEQRNAFIQSLKDDPSQSANLLAEAKKLNDAQAPKVDANCDIPTTASENLYFQGELKRR
                     WKKNFIAVSAANRFKKISSSGALDDDDKLIDTVDLEGGPGTQFAL"
     gene            1643..2269
                     /gene="tap"
                     /label=tap
     protein_bind    1646..1679
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     CDS             1712..1885
                     /codon_start=1
                     /product="IgG-binding unit of Staphylococcus aureus protein
                     A"
                     /label=ProtA
                     /translation="VDNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLL
                     AEAKKLNDAQAPK"
     CDS             1889..2059
                     /codon_start=1
                     /product="IgG-binding unit of Staphylococcus aureus protein
                     A"
                     /label=ProtA
                     /translation="DNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLLA
                     EAKKLNDAQAPK"
     CDS             2096..2116
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             2123..2200
                     /codon_start=1
                     /product="calmodulin-binding peptide"
                     /label=CBP
                     /note="derived from skeletal muscle myosin light chain 
                     kinase; binds calmodulin with nanomolar affinity in the 
                     presence of calcium"
                     /translation="KRRWKKNFIAVSAANRFKKISSSGAL"
     CDS             2201..2215
                     /codon_start=1
                     /product="enterokinase recognition and cleavage site"
                     /label=enterokinase site
                     /translation="DDDDK"
     primer_bind     complement(2221..2237)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             2280..3100
                     /codon_start=1
                     /gene="Xac amy"
                     /product="truncated alpha-amylase"
                     /label=Xac amy
                     /protein_id="ACM16584.1"
                     /translation="PGDVILHAFNWPYATVEARAKQIADAGYRKVLVAPAYRSEGSAWW
                     ARYQPQDIRLIDNPLGDTAAFKKMVQALASNGVETYADIVFNHMANEAATRSDLNYPGG
                     AVLSQYAANPGRYDSLRLFGTLQSNFLSASDFGPAQCISNYNDAYQVRNYRICGGGSDP
                     GLPDLLGNDWVVQQQRAYLQALKSMGVTGFRVDAAKHMTFDHLNRVFDAGIRSGVYVFG
                     EVITGGGTGNGDYDQFLAPYLQSTPHAAYDFPLFNAVRNAFAIGASMQQLP"
     gene            2280..3100
                     /gene="Xac amy"
                     /label=Xac amy
     primer_bind     complement(3110..3126)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      3267..3695
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     CDS             4039..4833
                     /codon_start=1
                     /gene="aphA"
                     /product="kanamycin resistance protein"
                     /label=aphA
                     /protein_id="ACM16583.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     gene            4039..4833
                     /gene="aphA"
                     /label=aphA
     CDS             4851..5711
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=bla
                     /protein_id="ACM16585.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            4851..5711
                     /gene="bla"
                     /label=bla
     rep_origin      5882..6470
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"