LOCUS       Exported                5159 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pmET32a, complete sequence.
ACCESSION   JX518291
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5159)
  AUTHORS   Lagzian M, Bassami MR, Dehghani H.
  TITLE     In vitro responses of chicken macrophage-like monocytes following 
            exposure to pathogenic and non-pathogenic E. coli ghosts loaded with
            a rational design of conserved genetic materials of influenza and 
            Newcastle disease viruses
  JOURNAL   Vet. Immunol. Immunopathol. 176, 5-17 (2016)
  PUBMED    27288852
REFERENCE   2  (bases 1 to 5159)
  AUTHORS   Lagzian M, Bassami MR, Dehghani H.
  TITLE     Nucleotide sequence of bacterial ghost creator vector
  JOURNAL   Unpublished
REFERENCE   3  (bases 1 to 5159)
  AUTHORS   Lagzian M, Bassami MR, Dehghani H.
  TITLE     Direct Submission
  JOURNAL   Submitted (23-AUG-2012) Biotechnology, Ferdowsi University of 
            Mashhad, Azadi sq., Mashhad, Khorasan-Razavi 91779-48974, Iran
REFERENCE   4  (bases 1 to 5159)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5159
                     /organism="Expression vector pmET32a"
                     /mol_type="other DNA"
                     /db_xref="taxon:1236202"
     rep_origin      1004..2282
     CDS             1050..1241
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     misc_feature    1343..1485
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(1671..2259)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2430..3290)
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=beta-lactamase
                     /note="ampillicin resistance"
                     /protein_id="AFT65570.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(3291..3395)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3422..3877
     terminator      3914..3961
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     regulatory      complement(3914..3960)
                     /regulatory_class="terminator"
     CDS             complement(4028..4045)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(4055..4768)
                     /codon_start=1
                     /product="temperature-sensitive lambda-repressor"
                     /label=temperature-sensitive lambda-repressor
                     /protein_id="AFT65571.1"
                     /translation="MSTKKKPLTQEQLEDARRLKAIYEKKKNELGLSQESVADKMGMGQ
                     SGVGALFNGINALNAYNAALLTKILKVSVEEFSPSIAREIYEMYEAVSMQPSLRSEYEY
                     PVFSHVQAGMFSPKLRTFTKGDAERWVSTTKKASDSAFWLEVEGNSMTAPTGSKPSFPD
                     GMLILVDPEQAVEPGDFCIARLGGDEFTFKKLIRDSGQVFLQPLNPQYPMIPCNESCSV
                     VGKVIASQWPEETFG"
     regulatory      4816..4843
                     /regulatory_class="promoter"
                     /note="lambda PR"
     RBS             4857..4865
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3 end of the 16S rRNA (Chen et al., 1994)"
     CDS             4872..5147
                     /codon_start=1
                     /product="E lysis protein"
                     /label=E lysis protein
                     /protein_id="AFT65572.1"
                     /translation="MVRWTLWDTLAFLLLLSLLLPSLLIMFIPSTFKRPVSSWKALNLR
                     KTLLMASSVRLKPLNCSRLPCVYAQETLTFLLTQKKTCVKNYVQKE"