LOCUS       Exported                5957 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Shuttle vector pBACOV-sfGFP, complete sequence.
ACCESSION   MG599121
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5957)
  AUTHORS   Heinze S, Kornberger P, Graetz C, Schwarz WH, Zverlov VV, Liebl W.
  TITLE     Conjugative transfer of a new broad host range expression vector to 
            various Bacillus species using a single protocol
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5957)
  AUTHORS   Heinze S, Kornberger P, Graetz C, Schwarz WH, Zverlov VV, Liebl W.
  TITLE     Direct Submission
  JOURNAL   Submitted (04-DEC-2017) Department for Microbiology, Technical 
            University of Munich, Emil-Ramann-Str. 4, Freising, Bavaria 85354, 
            Germany
REFERENCE   3  (bases 1 to 5957)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..5957
                     /organism="Shuttle vector pBACOV-sfGFP"
                     /mol_type="other DNA"
                     /db_xref="taxon:2184487"
     regulatory      complement(77..676)
                     /regulatory_class="replication_regulatory_region"
                     /note="ColE1 origin of replication, E. coli"
     rep_origin      complement(79..667)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(838..1698)
                     /codon_start=1
                     /gene="ampR"
                     /product="ampicillin resistance protein"
                     /label=ampR
                     /note="AmpR"
                     /protein_id="AWI97858.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(838..1698)
                     /gene="ampR"
                     /label=ampR
                     /note="for selection in Escherichia coli"
     promoter        complement(1699..1803)
                     /gene="bla"
                     /label=AmpR promoter
     regulatory      complement(1832..2353)
                     /regulatory_class="other"
                     /note="origin of transfer for conjugation, contains traJ 
                     and oriT"
     CDS             complement(1832..2116)
                     /codon_start=1
                     /gene="traJ"
                     /product="TraJ"
                     /label=traJ
                     /protein_id="AWI97859.1"
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGQQKAHYVNHHPNQVFWGRGAVKH"
     gene            complement(1832..2116)
                     /gene="traJ"
                     /label=traJ
                     /note="required for conjugation"
     regulatory      complement(2117..2353)
                     /regulatory_class="other"
                     /note="oriT (origin of transfer)"
     oriT            2149..2258
                     /note="incP origin of transfer"
     CDS             complement(2515..3276)
                     /codon_start=1
                     /gene="kanR"
                     /product="kanamycin resistance protein"
                     /label=kanR
                     /note="KanR"
                     /protein_id="AWI97860.1"
                     /translation="MNGPIIMTREERMKIVHEIKERILDKYGDDVKAIGVYGSLGRQTD
                     GPYSDIEMMCVMSTEEAEFSHEWTTGEWKVEVNFDSEEILLDYASQVESDWPLTHGQFF
                     SILPIYDSGGYLEKVYQTAKSVEAQTFHDAICALIVEELFEYAGKWRNIRVQGPTTFLP
                     SLTVQVAMAGAMLIGLHHRICYTTSASVLTEAVKQSDLPSGYDHLCQFVMSGQLSDSEK
                     LLESLENFWNGIQEWTERHGYIVDVSKRIPF"
     gene            complement(2515..3276)
                     /gene="kanR"
                     /label=kanR
                     /note="for selection in Bacillus"
     regulatory      complement(3445..4612)
                     /regulatory_class="replication_regulatory_region"
                     /note="pUB origin of replication for Bacillus"
     CDS             complement(3445..4449)
                     /codon_start=1
                     /gene="repB"
                     /product="RepB replication protein"
                     /label=repB
                     /note="from Enterococcus faecalis plasmid pAM-alpha-1"
                     /db_xref="GI:22652809"
                     /protein_id="AAN03827.1"
                     /translation="MGVSFNIMCPNSSIYSDEKSRVLVDKTKSGKVRPWREKKIANVDY
                     FELLHILEFKKAERVKDCAEILEYKQNRETGERKLYRVWFCKSRLCPMCNWRRAMKHGI
                     QSQKVVAEVIKQKPTVRWLFLTLTVKNVYDGEELNKSLSDMAQGFRRMMQYKKINKNLV
                     GFMRATEVTINNKDNSYNQHMHVLVCVEPTYFKNTENYVNQKQWIQFWKKAMKLDYDPN
                     VKVQMIRPKNKYKSDIQSAIDETAKYPVKDTDFMTDDEEKNLKRLSDLEEGLHRKRLIS
                     YGGLLKEIHKKLNLDDTEEGDLIHTDDDEKADEDGFSIIAMWNWERKNYFIKE"
     regulatory      4866..5031
                     /regulatory_class="promoter"
                     /note="aprE promoter from Bacillus subtilis"
     regulatory      4960..4969
                     /regulatory_class="minus_35_signal"
     regulatory      4987..4992
                     /regulatory_class="minus_10_signal"
     misc_feature    5057..5797
                     /note="sfGFP-His6; sfGFP with C-terminal hexahistidine tag;
                     super folder GFP"
     CDS             5777..5794
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"