LOCUS       40924_30765        5169 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Shuttle vector pMG14, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5169)
  AUTHORS   Gimpel M, Brantl S.
  TITLE     Construction of a modular plasmid family for chromosomal integration
            in Bacillus subtilis
  JOURNAL   J. Microbiol. Methods 91 (2), 312-317 (2012)
  PUBMED    22982324
REFERENCE   2  (bases 1 to 5169)
  AUTHORS   De Schamphelaire W, Olbrechts A, Meert J, Verhelst K, Roggeman 
            Fonseca M, Vanhoucke M, Beyaert R.
  TITLE     BCCM/LMBP Plasmid collection
  JOURNAL   Unpublished
REFERENCE   3  (bases 1 to 5169)
  AUTHORS   De Schamphelaire W.
  TITLE     Direct Submission
  JOURNAL   Submitted (02-FEB-2017) BCCM/LMBP, Universiteit Gent, 
            Technologiepark 927, 9052, BELGIUM
REFERENCE   4  (bases 1 to 5169)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 5169)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J. 
            Microbiol. Methods"; date: "2012"; volume: "91"; issue: "2"; pages: 
            "312-317"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 3; type: "Journal Article"; journalName: "Submitted 
            (02-FEB-2017) BCCM/LMBP, Universiteit Gent, Technologiepark 927, 
            9052, BELGIUM"
COMMENT     SGRef: number: 4; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5169
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      5..234
                     /label=cggR promoter
                     /note="cggR promoter"
                     /regulatory_class="promoter"
     misc_feature    262..302
                     /label=5' UTR of gapA
                     /note="5' UTR of gapA"
     CDS             311..334
                     /label=Strep-Tag II
                     /note="peptide that binds Strep-Tactin(R), an engineered 
                     form
                     of streptavidin"
     CDS             complement(470..1009)
                     /codon_start=1
                     /note="unnamed protein product; amyE fragment"
                     /protein_id="SJL86547.1"
                     /translation="MFAKRFKTSLLPLFAGFLLLFHLVLAGPAAASAETANKSNELTAP
                     SIKSGTILHAWNWSFNTLKHNMKDIHDAGYTAIQTSPINQVKEGNQGDKSMSNWYWLYQ
                     PTSYQIGNRYLGTEQEFKEMCAAAEEYGIKVIVDAVINHTTSDYAAISNEVKSIPNWTH
                     GNTQIKNWSDRWDVTQN"
     misc_feature    complement(1010..1125)
                     /label=5' UTR of amyE
                     /note="5' UTR of amyE"
     promoter        1229..1333
                     /label=AmpR promoter
     CDS             1334..2191
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      2365..2953
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    3241..3262
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        3277..3307
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3315..3331
                     /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     3339..3355
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(3396..4199)
                     /codon_start=1
                     /note="unnamed protein product; amyE fragment"
                     /protein_id="SJL86549.1"
                     /translation="STWMSDDDIRLGWAVIASRSGSTPLFFSRPEGGGNGVRFPGKSQI
                     GDRGSALFEDQAITAVNRFHNVMAGQPEELSNPNGNNQIFMNQRGSHGVVLANAGSSSV
                     SINTATKLPDGRYDNKAGAGSFQVNDGKLTGTINARSVAVLYPDDIAKAPHVFLENYKT
                     GVTHSFNDQLTITLRADANTTKAVYQINNGPETAFKDGDQFTIGKGDPFGKTYTIMLKG
                     TNSDGVTRTEKYSFVKRDPASAKTIGYQNPNHWSQVNAYIYKHDGS"
     CDS             complement(4289..5059)
                     /codon_start=1
                     /note="unnamed protein product; kanamycin/tobramycin
                     resistance (aadD)"
                     /protein_id="SJL86550.1"
                     /translation="MRIVNGPIIMTREERMKIVHEIKERILDKYGDDVKAIGVYGSLGR
                     QTDGPYSDIEMMCVMSTEEAEFSHEWTTGEWKVEVNFDSEEILLDYASQVESDWPLTHG
                     QFFSILPIYDSGGYLEKVYQTAKSVEAQTFHDAICALIVEELFEYAGKWRNIRVQGPTT
                     FLPSLTVQVAMAGAMLIGLHHRICYTTSASVLTEAVKQSDLPSGYDHLCQFVMSGQLSD
                     SEKLLESLENFWNGIQEWTERHGYIVDVSKRIPF"
ORIGIN
        1 aattctttgg ataaaatgaa aacgctttaa tgaaacagcc ctttctacgg gaagggctgt
       61 ttatattggg atgcaccatt tggcgctttc tgtataagat aaagatatat aggataaaat
      121 attgctggat aaaacgacgc ggcatgaaaa ctctgcgaat attgtcgatg aattggctct
      181 taacagttga ataaacaatt ccaccctgtt aaaataatta aagaaagcag aaatctgcag
      241 aagttattaa gggatgaata atccctcaat ataaatatct ctcacttatt taaaggagga
      301 aagcttcatg tggagccacc cgcagttcga aaaaggtacc cggggatcct ctagagtcct
      361 gagcgccggt cgctaccatt accagttggt ctggtgtcaa aaataataat aaccgggcag
      421 gccatgtctg cccgtatttc gcgtaaggaa atccattatg tactatttaa ttctgcgtga
      481 catcccatcg atcagaccag tttttaattt gtgtgtttcc atgtgtccag tttggaatac
      541 tcttaacctc attggaaatc gcggcataat cactggtggt atgattgatg accgcgtcaa
      601 caatgacctt tatgccatat tcttcagcgg ctgcacacat ttctttaaat tcttgttcag
      661 tacctaagta acggttgcca atttgatacg atgtcggctg atacagccag taccagttcg
      721 acatgctttt atctccttga ttcccttcct ttacttggtt aatcggagat gtctgaatgg
      781 ctgtatatcc tgcatcatga atatccttca tattgtgttt taacgtattg aacgaccaat
      841 tccatgcatg aagaatggtt ccgcttttga tcgacggtgc tgtaagctca ttcgatttgt
      901 tcgccgtttc agcactcgca gccgccggtc ctgccagaac caaatgaaac agcaataaaa
      961 atccagcgaa taacggcagt aaagaggttt tgaatcgttt tgcaaacatt cttgacactc
     1021 cttatttgat tttttgaaga cttacttcgg agtcaaaaat ccctcttact tcattcttcc
     1081 gcttcctcct ttcaaaccga tgtgaagact ggagaatttt gttaattctt gaagacgaaa
     1141 gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac
     1201 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     1261 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     1321 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     1381 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     1441 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     1501 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     1561 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     1621 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     1681 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     1741 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     1801 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     1861 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     1921 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     1981 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     2041 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     2101 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     2161 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     2221 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     2281 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     2341 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     2401 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     2461 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     2521 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     2581 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     2641 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     2701 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     2761 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     2821 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     2881 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     2941 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     3001 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     3061 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     3121 cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca
     3181 ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat
     3241 taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg
     3301 tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga
     3361 ttacgccaag ctaatcggcg gccgcatcga tgcaggctcc catcatgttt atagatataa
     3421 gcatttacct ggctccaatg attcggattt tgatagccga tggttttggc cgacgctgga
     3481 tctcttttaa caaaactgta tttctcggtc ctcgttacac catcactgtt cgttcctttt
     3541 aacatgatgg tgtatgtttt gccaaatgga tctccttttc cgattgtgaa ttgatctcca
     3601 tccttaaacg ccgtctctgg tccattattg atttgataaa cggcttttgt tgtattcgca
     3661 tctgcacgca aggtaatcgt cagttgatca ttgaaagaat gtgttacacc tgttttgtaa
     3721 ttctcaagga aaacatgagg cgcttttgca atatcatcag gataaagcac agctacagac
     3781 ctggcattga tcgtgcctgt cagtttacca tcgttcactt gaaatgaacc cgctccagct
     3841 ttattgtcat acctgccatc aggcaatttt gttgccgtat tgatagagac agaggatgaa
     3901 cctgcatttg ccagcacaac gccatgtgag ccgcgctgat tcataaatat ctggttgttt
     3961 ccattcgggt tcgagagttc ctcaggctgt ccagccatca cattgtgaaa tctattgacc
     4021 gcagtgatag cctgatcttc aaataaagca ctcccgcgat cgcctatttg gcttttcccc
     4081 gggaacctca caccatttcc gcctccctca ggtctggaaa agaaaagagg cgtactgcct
     4141 gaacgagaag ctatcaccgc ccagcctaaa cggatatcat catcgctcat ccatgtcgac
     4201 gcgccatgac agccatgata attactaata ctaggagaag ttaataaata cgtaaccaac
     4261 atgattaaca attattagag gtcatcgttc aaaatggtat gcgttttgac acatccacta
     4321 tatatccgtg tcgttctgtc cactcctgaa tcccattcca gaaattctct agcgattcca
     4381 gaagtttctc agagtcggaa agttgaccag acattacgaa ctggcacaga tggtcataac
     4441 ctgaaggaag atctgattgc ttaactgctt cagttaagac cgaagcgctc gtcgtataac
     4501 agatgcgatg atgcagacca atcaacatgg cacctgccat tgctacctgt acagtcaagg
     4561 atggtagaaa tgttgtcggt ccttgcacac gaatattacg ccatttgcct gcatattcaa
     4621 acagctcttc tacgataagg gcacaaatcg catcgtggaa cgtttgggct tctaccgatt
     4681 tagcagtttg atacactttc tctaagtatc cacctgaatc ataaatcggc aaaatagaga
     4741 aaaattgacc atgtgtaagc ggccaatctg attccacctg agatgcataa tctagtagaa
     4801 tctcttcgct atcaaaattc acttccacct tccactcacc ggttgtccat tcatggctga
     4861 actctgcttc ctctgttgac atgacacaca tcatctcaat atccgaatag ggcccatcag
     4921 tctgacgacc aagagagcca taaacaccaa tagccttaac atcatcccca tatttatcca
     4981 atattcgttc cttaatttca tgaacaatct tcattctttc ttctctagtc attattattg
     5041 gtccattcac tattctcatt cccttttcag ataattttag atttgctttt ctaaataaga
     5101 atatttggag agcaccgttc ttattcagct attaaaccca ttatatcggg tttttgaggg
     5161 gatttcaag
//
