LOCUS       Exported                7214 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Shuttle vector pMK3, complete sequence.
ACCESSION   EU549779
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7214)
  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  JOURNAL   Unpublished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 7214)
  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 7214)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7214
                     /organism="Shuttle vector pMK3"
                     /mol_type="other DNA"
                     /note="constructed from Staphylococcus aureus plasmid 
                     pUB110 and Escherichia coli cloning vector pUC9"
                     /db_xref="taxon:515189"
     misc_feature    1..383
                     /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..1364
                     /codon_start=1
                     /gene="lacZ-mob"
                     /product="LacZ-Mob fusion protein"
                     /label=lacZ-mob
                     /note="fusion between beta galactosidase alpha peptide 
                     fragment and Mob protein fragment; complementation of 
                     beta-lactamase delta-M15 fragment to allow enzymatic 
                     activity; created by fusion between Staphylococcus aureus 
                     plasmid pUB110 and Escherichia coli cloning vector pUC9 
                     during construction"
                     /protein_id="ACB41737.1"
                     /translation="MTMITPSLAAGRRIPGNSLAVVLQRRDWENPGVTQLNRLAAHPPF
                     ASWRNSEEARTDPGEQKRFFEESYKLFSERYGKQNIAYATVHNDEQTPHMHLGVVPMRD
                     GKLQGKNVFNRQELLWLQDKFPEHMKKQGFELKRGERGSDRKHIETAKFKKQTLEKEID
                     FLEKNLAVKKDEWTAYSDKVKSDLEVPAKRHMKSVEVPTGEKSMFGLGKEIMKTEKKPT
                     KNVVISERDYKNLVTAARDNDRLKQHVRNLMSTDMAREYKKLSKEHGQVKEKYSGLVER
                     FNENVNDYNELLEENKSLKSKISDLKRDVSLIYESTKEFLKERTDGLKAFKNVFKGFVD
                     KVKDKTAQFQEKHDLEPKKNEFELTHNREVKKERSRDQGMSL"
     gene            216..1364
                     /gene="lacZ-mob"
                     /label=lacZ-mob
     primer_bind     complement(269..285)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    384..4932
                     /label=derived from pC194
                     /note="derived from pC194"
     CDS             1588..2592
                     /codon_start=1
                     /gene="repB"
                     /product="replication initiation protein"
                     /label=repB
                     /note="RepB; initiation of plasmid replication; derived 
                     from Staphylococcus aureus plasmid pUB110"
                     /protein_id="ACB41740.1"
                     /translation="MGVSFNIMCPNSSIYSDEKSRVLVDKTKSGKVRPWREKKIANVDY
                     FELLHILEFKKAERVKDCAEILEYKQNRETGERKLYRVWFCKSRLCPMCNWRRAMKHGI
                     QSQKVVAEVIKQKPTVRWLFLTLTVKNVYDGEELNKSLSDMAQGFRRMMQYKKINKNLV
                     GFMRATEVTINNKDNSYNQHMHVLVCVEPTYFKNTENYVNQKQWIQFWKKAMKLDYDPN
                     VKVQMIRPKNKYKSDIQSAIDETAKYPVKDTDFMTDDEEKNLKRLSDLEEGLHRKRLIS
                     YGGLLKEIHKKLNLDDTEEGDLIHTDDDEKADEDGFSIIAMWNWERKNYFIKE"
     gene            1588..2592
                     /gene="repB"
                     /label=repB
     CDS             2752..3522
                     /codon_start=1
                     /gene="kan"
                     /product="kanamycin nucleotidyltransferase"
                     /EC_number="2.7.7.-"
                     /label=kan
                     /note="kanamycin and neomycin resistance; derived from 
                     Staphylococcus aureus plasmid pUB110"
                     /protein_id="ACB41738.1"
                     /translation="MRIVNGPIIMTREERMKIVHEIKERILDKYGDDVKAIGVYGSLGR
                     QTDGPYSDIEMMCVMSTEEAEFSHEWTTGEWKVEVNFDSEEILLDYASQVESDWPLTHG
                     QFFSILPIYDSGGYLEKVYQTAKSVEAQTFHDAICALIVEELFEYAGKWRNIRVQGPTT
                     FLPSLTVQVAMAGAMLIGLHHRICYTTSASVLTEAVKQSDLPSGYDHLCQFVMSGQLSD
                     SEKLLESLENFWNGIQEWTERHGYIVDVSKRIPF"
     gene            2752..3522
                     /gene="kan"
                     /label=kan
     CDS             3745..4143
                     /codon_start=1
                     /product="bleomycin resistance protein"
                     /EC_number="4.4.1.5"
                     /label=bleomycin resistance protein
                     /note="glyoxylase; bleomycin resistance; derived from 
                     Staphylococcus aureus plasmid pUB110"
                     /protein_id="ACB41739.1"
                     /translation="MLQSIPALPVGDIKKSIGFYCDKLGFTLVHHEDGFAVLMCNEVRI
                     HLWEASDEGWRSRSNDSPVCTGAESFIAGTASCRIEVEGIDELYQHIKPLGILHPNTSL
                     KDQWWDERDFAVIDPDNNLISFFQQIKS"
     misc_feature    4933..7214
                     /label=derived from pUC9
                     /note="derived from pUC9"
     promoter        5308..5412
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5413..6273
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /EC_number="3.5.2.6"
                     /label=bla
                     /note="ampicillin resistance; derived from Escherichia coli
                     cloning vector pUC9"
                     /protein_id="ACB41741.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            5413..6273
                     /gene="bla"
                     /label=bla
     rep_origin      6444..7032
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"