LOCUS       40924_31005        5585 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Shuttle vector pMK4, complete sequence.
ACCESSION   .
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  JOURNAL   Unpublished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)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5585)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (06-MAR-2008) Bacillus Genetic Stock Center, The Ohio State 
            University, 484 W 12th Ave, Columbus, OH 43210, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5585
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    106..127
                     /label=CAP binding site
                     /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
                     /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"
     primer_bind     complement(269..285)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        759..863
                     /label=AmpR promoter
     CDS             864..1721
                     /label=AmpR
                     /note="beta-lactamase"
     CDS             complement(2460..2474)
                     /label=enterokinase site
                     /note="enterokinase recognition and cleavage site"
     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
     CDS             3965..4612
                     /gene="cat"
                     /label=cat
                     /note="Chloramphenicol acetyltransferase from
                     Staphylococcus aureus. Accession#: P00485"
     rep_origin      4815..5403
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
       61 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct
      121 cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat
      181 tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg ccaagcttgg
      241 ctgcaggtcg acggatcccc gggaattcac tggccgtcgt tttacaacgt cgtgactggg
      301 aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc
      361 gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg
      421 aatggcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatat
      481 ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc
      541 caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag
      601 ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg
      661 cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg
      721 tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
      781 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
      841 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
      901 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
      961 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     1021 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     1081 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     1141 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     1201 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     1261 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     1321 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     1381 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     1441 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     1501 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     1561 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     1621 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     1681 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     1741 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     1801 agatccatat ccttcttttt ctgaaccgac ttctcctttt tcgcttcttt attccaattg
     1861 ctttattgac gttgagcctc ggaaccctta acaatcccaa aacttgtcga atggtcggct
     1921 taatagctca cgctatgccg acattcgtct gcaagtttag ttaagggttc ttctcaacgc
     1981 acaataaatt ttctcggcat aaatgcgtgg tctaattttt atttttaata accttgatag
     2041 caaaaaatgc cattccaata caaaaccaca tacctataat cgataaccac ataacagtca
     2101 taaaaccact cctttttaac aaactttatc acaagaaata tttaaatttt aaatgccttt
     2161 attttgaatt ttaaggggca ttttaaagat ttaggggtaa atcatatagt tttatgccta
     2221 aaaacctaca gaagctttta aaaagcaaat atgagccaaa taaatatatt ctaattctac
     2281 aaacaaaaat ttgagcaaat tcagtgtcga ttttttaaga cactgcccag ttacatgcaa
     2341 attaaaattt tcatgatttt ttatagttcc taacagggtt aaaatttgta taacgaaagt
     2401 ataatgttta tataacgtta gtataataaa gcattttaac attatacttt tgataatcgt
     2461 ttatcgtcgt catcacaata acttttaaaa tactcgtgca taattcaaca gctgacctcc
     2521 caataactac atggtgttat cgggaggtca gctgttagca cttatatttt gttattgttc
     2581 ttcctcgatt tcgtctatca ttttgtgatt aatttctctt ttttcttgtt ctgttaagtc
     2641 ataaagttca ctagctaaat actctttttg tttccaaata taaaaaattt gatagatata
     2701 ttcggttgga tcaatttctt ttaagtaatc taaatcccca ttttttaatt tctttttagc
     2761 ctctttaaat aatcctgaat aaactaatac ctgtttacct ttaagtgatt tataaaatgc
     2821 atcaaagact ttttgattta ttaaataatc actatcttta ccagaatact tagccatttc
     2881 atataattct ttattattat tttgtcttat tttttgaact tgaacttgtg ttatttctga
     2941 aatgcccgtt acatcacgcc ataaatctaa ccattcttgt tggctaatat aatatctttt
     3001 atctgtgaaa tacgatttat ttactgcaat taacacatga aaatgaggat tataatcatc
     3061 tcttttttta ttatatgtaa tctctaactt acgaacatat ccctttataa cactacctac
     3121 tttttttctc tttataagtt ttctaaaaga attattataa cgttttattt cattttctaa
     3181 ttcatcactc attacattag gtgtagtcaa agttaaaaag ataaactcct ttttctcttg
     3241 ctgcttaata tattgcatca tcaaagataa acccaatgca tcttttctag cttttctcca
     3301 agcacagaca ggacaaaatc gatttttaca agaattagct ttatataatt tctgtttttc
     3361 taaagtttta tcagctacaa aagacagaaa tgtattgcaa tcttcaacta aatccatttg
     3421 attctctcca atatgacgtt taataaattt ctgaaatact tgatttcttt gttttttctc
     3481 agtatacttt tccatgttat aacacataaa aacaacttag ttttcacaaa ctatgacaat
     3541 aaaaaaagtt gctttttccc ctttctatgt atgtttttta ctagtcattt aaaacgatac
     3601 attaataggt acgaaaaagc aacttttttt gcgcttaaaa ccagtcatac caataactta
     3661 agggtaacta gcctcgccgg caatagttac ccttattatc aagataagaa agaaaaggat
     3721 ttttcgctac gctcaaatcc tttaaaaaaa cacaaaagac cacatttttt aatgtggtct
     3781 tttattcttc aactaaagca cccattagtt caacaaacga aaattggata aagtgggata
     3841 tttttaaaat atatatttat gttacagtaa tattgacttt taaaaaagga ttgattctaa
     3901 tgaagaaagc agacaagtaa gcctcctaaa ttcactttag ataaaaattt aggaggcata
     3961 tcaaatgaac tttaataaaa ttgatttaga caattggaag agaaaagaga tatttaatca
     4021 ttatttgaac caacaaacga cttttagtat aaccacagaa attgatatta gtgttttata
     4081 ccgaaacata aaacaagaag gatataaatt ttaccctgca tttattttct tagtgacaag
     4141 ggtgataaac tcaaatacag cttttagaac tggttacaat agcgacggag agttaggtta
     4201 ttgggataag ttagagccac tttatacaat ttttgatggt gtatctaaaa cattctctgg
     4261 tatttggact cctgtaaaga atgacttcaa agagttttat gatttatacc tttctgatgt
     4321 agagaaatat aatggttcgg ggaaattgtt tcccaaaaca cctatacctg aaaatgcttt
     4381 ttctctttct attattccat ggacttcatt tactgggttt aacttaaata tcaataataa
     4441 tagtaattac cttctaccca ttattacagc aggaaaattc attaataaag gtaattcaat
     4501 atatttaccg ctatctttac aggtacatca ttctgtttgt gatggttatc atgcaggatt
     4561 gtttatgaac tctattcagg aattgtcaga taggcctaat gactggcttt tataatatga
     4621 gataatgccg actgtacttt ttacagtcgg ttttctaatg tcactaacct gccccgttag
     4681 ttgaagaagg tttttatatt acagctccag atctaggtga agatcctttt tgataatctc
     4741 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     4801 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     4861 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     4921 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     4981 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     5041 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     5101 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     5161 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     5221 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     5281 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     5341 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     5401 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     5461 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     5521 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     5581 gaaga
//
