LOCUS       Exported                5169 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Shuttle vector pMG14, complete sequence.
ACCESSION   LT726805
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  JOURNAL   Unpublished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)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5169
                     /organism="Shuttle vector pMG14"
                     /mol_type="other DNA"
                     /note="BCCM/LMBP Plasmid collection (Ghent 
                     University,Belgium) accession number LMBP 8465."
                     /db_xref="taxon:1945224"
     regulatory      5..234
                     /regulatory_class="promoter"
                     /note="cggR promoter"
     misc_feature    262..302
                     /label=5 UTR of gapA
                     /note="5 UTR of gapA"
     misc_feature    311..334
                     /label=Strep-tag II
                     /note="Strep-tag II"
     CDS             311..334
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     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
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1334..2194
                     /codon_start=1
                     /note="unnamed protein product; ampicillin resistance"
                     /protein_id="SJL86548.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      2365..2953
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      2735..3137
                     /label=pMB1 ori
                     /note="pMB1 ori"
     regulatory      3229..3312
                     /regulatory_class="promoter"
                     /note="lac promoter"
     protein_bind    3241..3262
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3277..3307
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     misc_feature    3313..3350
                     /label=5 UTR of lacZ, incl. RBS
                     /note="5 UTR of lacZ, incl. RBS"
     protein_bind    3315..3331
                     /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     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"