LOCUS       Exported                3565 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pMH, complete sequence.
ACCESSION   AY262822
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3565)
  AUTHORS   Gortchakov AA, Kaltenhauser J, Eggert H, Saumweber H.
  TITLE     pRich, a novel convenient Escherichia coli protein expression vector
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 3565)
  AUTHORS   Gortchakov AA, Kaltenhauser J, Eggert H, Saumweber H.
  TITLE     Direct Submission
  JOURNAL   Submitted (27-MAR-2003) Lab. Molecular Cytogenetics, The Institute 
            of Cytology and Genetics, SD RAS, Lavrentjeva, 10, Novosibirsk 
            630090, Russia
REFERENCE   3  (bases 1 to 3565)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..3565
                     /organism="Expression vector pMH"
                     /mol_type="genomic DNA"
                     /note="assembled from pQE30 (Qiagen) and pET3a (Stratagene)
                     with a dual (myc)3-(His)6 tag introduced"
                     /db_xref="taxon:228099"
     promoter        26..44
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      39..45
                     /regulatory_class="promoter"
                     /note="T7 promoter"
     RBS             76..98
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     regulatory      91..102
                     /regulatory_class="ribosome_binding_site"
     misc_feature    106..243
                     /label=(myc)3-(his)6-epitope
                     /note="(myc)3-(his)6-epitope"
     CDS             118..147
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             154..183
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             190..219
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             223..240
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     regulatory      237..246
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     misc_feature    242..257
                     /label=polylinker
                     /note="polylinker"
     regulatory      313..407
                     /regulatory_class="terminator"
                     /note="lambda t0 transcription terminator region"
     terminator      313..407
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             451..1110
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     regulatory      1169..1267
                     /regulatory_class="terminator"
                     /note="rrnB T1 transcription terminator region"
     terminator      1175..1261
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     misc_feature    1416..1556
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(1742..2330)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2501..3361)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /function="confers resistance to ampicillin"
                     /label=bla
                     /protein_id="AAP06957.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(2501..3361)
                     /gene="bla"
                     /label=bla
     promoter        complement(3362..3466)
                     /gene="bla"
                     /label=AmpR promoter