LOCUS       40924_30975        7214 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Shuttle vector pMK3, complete sequence.
ACCESSION   .
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  JOURNAL   Unpublished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)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7214)
  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            "Unpublished"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..7214
                     /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"
     CDS             1588..2589
                     /codon_start=1
                     /label=repB
                     /note="RepB replication protein"
                     /translation="MGVSFNIMCPNSSIYSDEKSRVLVDKTKSGKVRPWREKKIANVDY
                     FELLHILEFKKAERVKDCAEILEYKQNRETGERKLYRVWFCKSRLCPMCNWRRAMKHGI
                     QSQKVVAEVIKQKPTVRWLFLTLTVKNVYDGEELNKSLSDMAQGFRRMMQYKKINKNLV
                     GFMRATEVTINNKDNSYNQHMHVLVCVEPTYFKNTENYVNQKQWIQFWKKAMKLDYDPN
                     VKVQMIRPKNKYKSDIQSAIDETAKYPVKDTDFMTDDEEKNLKRLSDLEEGLHRKRLIS
                     YGGLLKEIHKKLNLDDTEEGDLIHTDDDEKADEDGFSIIAMWNWERKNYFIKE"
     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"
     promoter        5308..5412
                     /label=AmpR promoter
     CDS             5413..6270
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      6444..7032
                     /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 accgatccag gagaacaaaa acgatttttt gaggaaagtt
      421 ataaattatt ttccgaacga tatggcaagc aaaatattgc ttatgcaaca gttcataatg
      481 atgagcaaac ccctcacatg catttaggtg ttgtgcctat gcgtgatgga aaactgcaag
      541 gaaaaaatgt gtttaatcgt caagaactgt tatggctaca agataaattc cccgagcata
      601 tgaaaaaaca gggttttgag ttgaagcgtg gtgaacgtgg ctctgaccgt aaacatattg
      661 agacagctaa atttaaaaaa caaactttgg aaaaagagat tgattttcta gaaaaaaatt
      721 tagcagttaa aaaagatgaa tggactgctt atagcgataa agttaaatca gatttagaag
      781 taccagcgaa acgacacatg aaaagtgttg aagtgccaac gggtgaaaag tccatgtttg
      841 gtttgggaaa agaaataatg aaaacagaaa agaaaccaac caaaaatgtt gttatatcgg
      901 agcgtgatta taaaaactta gtgactgctg cgagagataa cgataggtta aaacagcatg
      961 ttagaaatct catgagtact gatatggcga gagaatataa aaaattaagt aaagaacatg
     1021 ggcaagttaa agaaaaatat agtggtcttg tagagcgatt taatgaaaat gtaaatgatt
     1081 ataatgagtt gcttgaagaa aacaagtctt taaagtctaa aataagcgat ttaaagcgtg
     1141 atgtgagttt aatctatgaa agcactaagg aattccttaa ggaacgtaca gacggcttaa
     1201 aagcctttaa aaacgttttt aaggggtttg tagacaaggt aaaggataaa acagcacaat
     1261 tccaagaaaa acacgattta gaacctaaaa agaacgaatt tgaactaact cataaccgag
     1321 aggtaaaaaa agaacgaagt cgagatcagg gaatgagttt ataaaataaa aaaagcacct
     1381 gaaaaggtgt ctttttttga tggttttgaa cttgttcttt cttatcttga tacatataga
     1441 aataacgtca tttttatttt agttgctgaa aggtgcgttg aagtgttggt atgtatgtgt
     1501 tttaaagtat tgaaaaccct taaaattggt tgcacagaaa aaccccatct gttaaagtta
     1561 taagtgacta aacaaataac taaatagatg ggggtttctt ttaatattat gtgtcctaat
     1621 agtagcattt attcagatga aaaatcaagg gttttagtgg acaagacaaa aagtggaaaa
     1681 gtgagaccat ggagagaaaa gaaaatcgct aatgttgatt actttgaact tctgcatatt
     1741 cttgaattta aaaaggctga aagagtaaaa gattgtgctg aaatattaga gtataaacaa
     1801 aatcgtgaaa caggcgaaag aaagttgtat cgagtgtggt tttgtaaatc caggctttgt
     1861 ccaatgtgca actggaggag agcaatgaaa catggcattc agtcacaaaa ggttgttgct
     1921 gaagttatta aacaaaagcc aacagttcgt tggttgtttc tcacattaac agttaaaaat
     1981 gtttatgatg gcgaagaatt aaataagagt ttgtcagata tggctcaagg atttcgccga
     2041 atgatgcaat ataaaaaaat taataaaaat cttgttggtt ttatgcgtgc aacggaagtg
     2101 acaataaata ataaagataa ttcttataat cagcacatgc atgtattggt atgtgtggaa
     2161 ccaacttatt ttaagaatac agaaaactac gtgaatcaaa aacaatggat tcaattttgg
     2221 aaaaaggcaa tgaaattaga ctatgatcca aatgtaaaag ttcaaatgat tcgaccgaaa
     2281 aataaatata aatcggatat acaatcggca attgacgaaa ctgcaaaata tcctgtaaag
     2341 gatacggatt ttatgaccga tgatgaagaa aagaatttga aacgtttgtc tgatttggag
     2401 gaaggtttac accgtaaaag gttaatctcc tatggtggtt tgttaaaaga aatacataaa
     2461 aaattaaacc ttgatgacac agaagaaggc gatttgattc atacagatga tgacgaaaaa
     2521 gccgatgaag atggattttc tattattgca atgtggaatt gggaacggaa aaattatttt
     2581 attaaagagt agttcaacaa acgggccagt ttgttgaaga ttagatgcta taattgttat
     2641 taaaaggatt gaaggatgct taggaagacg agttattaat agctgaataa gaacggtgct
     2701 ctccaaatat tcttatttag aaaagcaaat ctaaaattat ctgaaaaggg aatgagaata
     2761 gtgaatggac caataataat gactagagaa gaaagaatga agattgttca tgaaattaag
     2821 gaacgaatat tggataaata tggggatgat gttaaggcta ttggtgttta tggctctctt
     2881 ggtcgtcaga ctgatgggcc ctattcggat attgagatga tgtgtgtcat gtcaacagag
     2941 gaagcagagt tcagccatga atggacaacc ggtgagtgga aggtggaagt gaattttgat
     3001 agcgaagaga ttctactaga ttatgcatct caggtggaat cagattggcc gcttacacat
     3061 ggtcaatttt tctctatttt gccgatttat gattcaggtg gatacttaga gaaagtgtat
     3121 caaactgcta aatcggtaga agcccaaacg ttccacgatg cgatttgtgc ccttatcgta
     3181 gaagagctgt ttgaatatgc aggcaaatgg cgtaatattc gtgtgcaagg accgacaaca
     3241 tttctaccat ccttgactgt acaggtagca atggcaggtg ccatgttgat tggtctgcat
     3301 catcgcatct gttatacgac gagcgcttcg gtcttaactg aagcagttaa gcaatcagat
     3361 cttccttcag gttatgacca tctgtgccag ttcgtaatgt ctggtcaact ttccgactct
     3421 gagaaacttc tggaatcgct agagaatttc tggaatggga ttcaggagtg gacagaacga
     3481 cacggatata tagtggatgt gtcaaaacgc ataccatttt gaacgatgac ctctaataat
     3541 tgttaatcat gttggttacg tatttattaa cttctcctag tattagtaat tatcatggct
     3601 gtcatggcgc attaacggaa taaagggtgt gcttaaatcg ggccattttg cgtaataaga
     3661 aaaaggatta attatgagcg aattgaatta ataataaggt aatagattta cattagaaaa
     3721 tgaaagggga ttttatgcgt gagaatgtta cagtctatcc cggcattgcc agtcggggat
     3781 attaaaaaga gtataggttt ttattgcgat aaactaggtt tcactttggt tcaccatgaa
     3841 gatggattcg cagttctaat gtgtaatgag gttcggattc atctatggga ggcaagtgat
     3901 gaaggctggc gctctcgtag taatgattca ccggtttgta caggtgcgga gtcgtttatt
     3961 gctggtactg ctagttgccg cattgaagta gagggaattg atgaattata tcaacatatt
     4021 aagcctttgg gcattttgca ccccaataca tcattaaaag atcagtggtg ggatgaacga
     4081 gactttgcag taattgatcc cgacaacaat ttgattagct tttttcaaca aataaaaagc
     4141 taaaatctat tattaatctg ttcagcaatc gggcgcgatt gctgaataaa agatacgaga
     4201 gacctctctt gtatcttttt tattttgagt ggttttgtcc gttacactag aaaaccgaaa
     4261 gacaataaaa attttattct tgctgagtct ggctttcggt aagctagaca aaacggacaa
     4321 aataaaaatt ggcaagggtt taaaggtgga gattttttga gtgatcttct caaaaaatac
     4381 tacctgtccc ttgctgattt ttaaacgagc acgagagcaa aacccccctt tgctgaggtg
     4441 gcagagggca ggtttttttg tttctttttt ctcgtaaaaa aaagaaaggt cttaaaggtt
     4501 ttatggtttt ggtcggcact gccgacagcc tcgcagagca cacactttat gaatataaag
     4561 tatagtgtgt tatactttac ttggaagtgg ttgccggaaa gagcgaaaat gcctcacatt
     4621 tgtgccacct aaaaaggagc gatttacata tgagttatgc agtttgtaga atgcaaaaag
     4681 tgaaatcagc tggactaaaa ggcatgcaat ttcataatca aagagagcga aaaagtagaa
     4741 cgaatgatga tattgaccat gagcgaacac gtgaaaatta tgatttgaaa aatgataaaa
     4801 atattgatta caacgaacgt gtcaaagaaa ttattgaatc acaaaaaaac aggtacaaga
     4861 aaaacgagga aagatgctgt tcttgtaaat gagttgctag taacatctga ccgagatttt
     4921 tttgagcaac tggatcgccc ttcccaacag ttgcgcagcc tgaatggcga atggcgcctg
     4981 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatatg gtgcactctc
     5041 agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct
     5101 gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc
     5161 tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag
     5221 ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg
     5281 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     5341 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     5401 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     5461 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     5521 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     5581 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     5641 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     5701 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     5761 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     5821 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     5881 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     5941 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     6001 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     6061 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     6121 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     6181 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     6241 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     6301 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     6361 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     6421 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     6481 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     6541 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg
     6601 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     6661 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     6721 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     6781 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     6841 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     6901 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     6961 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     7021 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     7081 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     7141 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     7201 gaggaagcgg aaga
//
