LOCUS       Exported                5585 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Shuttle vector pMK4, complete sequence.
ACCESSION   EU549778
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5585)
  AUTHORS   Zeigler DR.
  TITLE     pMK3 and pMK4 revisited: nucleotide sequences of first generation 
            Bacillus subtilis - E. coli shuttle vectors
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5585)
  AUTHORS   Zeigler DR.
  TITLE     Direct Submission
  JOURNAL   Submitted (06-MAR-2008) Bacillus Genetic Stock Center, The Ohio 
            State University, 484 W 12th Ave, Columbus, OH 43210, USA
REFERENCE   3  (bases 1 to 5585)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5585
                     /organism="Shuttle vector pMK4"
                     /mol_type="other DNA"
                     /note="constructed from Staphylococcus aureus plasmid pC194
                     and Escherichia coli cloning vector pUC9"
                     /db_xref="taxon:515188"
     misc_feature    1..1801
                     /label=derived from pUC9
                     /note="derived from pUC9"
     protein_bind    106..127
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        142..172
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    180..196
                     /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     204..220
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             216..518
                     /codon_start=1
                     /gene="lacZ-alpha"
                     /product="beta-galactosidase alpha peptide"
                     /label=lacZ-alpha
                     /note="complementation of beta-lactamase delta-M15 fragment
                     to allow enzymatic activity; derived from pUC9"
                     /protein_id="ACB41733.1"
                     /translation="MTMITPSLAAGRRIPGNSLAVVLQRRDWENPGVTQLNRLAAHPPF
                     ASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICSDAA"
     gene            216..518
                     /gene="lacZ-alpha"
                     /label=lacZ-alpha
     primer_bind     complement(269..285)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        759..863
                     /gene="bla"
                     /label=AmpR promoter
     CDS             864..1724
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /EC_number="3.5.2.6"
                     /label=bla
                     /note="ampicillin resistance; derived from pUC9"
                     /protein_id="ACB41734.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            864..1724
                     /gene="bla"
                     /label=bla
     misc_feature    1802..2708
                     /label=derived from pC194
                     /note="derived from pC194"
     CDS             complement(2460..2474)
                     /codon_start=1
                     /product="enterokinase recognition and cleavage site"
                     /label=enterokinase site
                     /translation="DDDDK"
     CDS             complement(2572..3507)
                     /codon_start=1
                     /gene="repC"
                     /product="replication initiation protein"
                     /label=repC
                     /note="RepC; initiation of plasmid replication; derived 
                     from Staphylococcus aureus plasmid pC194"
                     /protein_id="ACB41735.1"
                     /translation="MCYNMEKYTEKKQRNQVFQKFIKRHIGENQMDLVEDCNTFLSFVA
                     DKTLEKQKLYKANSCKNRFCPVCAWRKARKDALGLSLMMQYIKQQEKKEFIFLTLTTPN
                     VMSDELENEIKRYNNSFRKLIKRKKVGSVIKGYVRKLEITYNKKRDDYNPHFHVLIAVN
                     KSYFTDKRYYISQQEWLDLWRDVTGISEITQVQVQKIRQNNNKELYEMAKYSGKDSDYL
                     INQKVFDAFYKSLKGKQVLVYSGLFKEAKKKLKNGDLDYLKEIDPTEYIYQIFYIWKQK
                     EYLASELYDLTEQEKREINHKMIDEIEEEQ"
     gene            complement(2572..3507)
                     /gene="repC"
                     /label=repC
     misc_feature    2709..4719
                     /label=derived from pC194
                     /note="derived from pC194"
     CDS             3965..4615
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol O-acetyltransferase"
                     /EC_number="2.3.1.28"
                     /label=cat
                     /note="chloramphenicol resistance; derived from 
                     Staphylococcus aureus plasmid pC194"
                     /protein_id="ACB41736.1"
                     /translation="MNFNKIDLDNWKRKEIFNHYLNQQTTFSITTEIDISVLYRNIKQE
                     GYKFYPAFIFLVTRVINSNTAFRTGYNSDGELGYWDKLEPLYTIFDGVSKTFSGIWTPV
                     KNDFKEFYDLYLSDVEKYNGSGKLFPKTPIPENAFSLSIIPWTSFTGFNLNINNNSNYL
                     LPIITAGKFINKGNSIYLPLSLQVHHSVCDGYHAGLFMNSIQELSDRPNDWLL"
     gene            3965..4615
                     /gene="cat"
                     /label=cat
     misc_feature    4720..5585
                     /label=derived from pUC9
                     /note="derived from pUC9"
     rep_origin      4815..5403
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"