LOCUS       Exported                4993 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pmET32b, complete sequence.
ACCESSION   KF561236
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4993)
  AUTHORS   Ebrahimi Nik H, Bassami MR, Rad M, Mohri M, Hosseini Shahidi R.
  TITLE     Nucleotide sequence of bacterial ghost creator vector 2
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4993)
  AUTHORS   Ebrahimi Nik H, Bassami MR, Rad M, Mohri M, Hosseini Shahidi R.
  TITLE     Direct Submission
  JOURNAL   Submitted (20-AUG-2013) Pathobiology, Biotechnology, Ferdowsi 
            Pardis, Azadi Sq., Mashhad, Khorasan Razavi 9177948974, Iran
REFERENCE   3  (bases 1 to 4993)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4993
                     /organism="Expression vector pmET32b"
                     /mol_type="other DNA"
                     /db_xref="taxon:1425343"
     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"
     promoter        2347..2375
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             2564..3097
                     /codon_start=1
                     /gene="aacC1"
                     /product="aminoglycoside acetyltransferase"
                     /function="gentamycin resistance"
                     /label=aacC1
                     /protein_id="AHA82377.1"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     gene            2564..3097
                     /gene="aacC1"
                     /label=aacC1
     rep_origin      3256..3711
     terminator      3748..3795
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     regulatory      complement(3748..3794)
                     /regulatory_class="terminator"
     CDS             complement(3862..3879)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(3889..4602)
                     /codon_start=1
                     /product="temperature-sensitive lambda-repressor protein"
                     /label=temperature-sensitive lambda-repressor protein
                     /protein_id="AHA82379.1"
                     /translation="MSTKKKPLTQEQLEDARRLKAIYEKKKNELGLSQESVADKMGMGQ
                     SGVGALFNGINALNAYNAALLTKILKVSVEEFSPSIAREIYEMYEAVSMQPSLRSEYEY
                     PVFSHVQAGMFSPKLRTFTKGDAERWVSTTKKASDSAFWLEVEGNSMTAPTGSKPSFPD
                     GMLILVDPEQAVEPGDFCIARLGGDEFTFKKLIRDSGQVFLQPLNPQYPMIPCNESCSV
                     VGKVIASQWPEETFG"
     regulatory      4650..4677
                     /regulatory_class="promoter"
                     /note="lambda PR"
     RBS             4691..4699
                     /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             4706..4981
                     /codon_start=1
                     /product="E lysis protein"
                     /label=E lysis protein
                     /protein_id="AHA82378.1"
                     /translation="MVRWTLWDTLAFLLLLSLLLPSLLIMFIPSTFKRPVSSWKALNLR
                     KTLLMASSVRLKPLNCSRLPCVYAQETLTFLLTQKKTCVKNYVQKE"