LOCUS       Exported                8965 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Delivery vector pIEFBPR, complete sequence.
ACCESSION   DQ486035
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8965)
  AUTHORS   Zhang XZ, Yan X, Cui ZL, Hong Q, Li SP.
  TITLE     mazF, a novel counter-selectable marker for unmarked chromosomal 
            manipulation in Bacillus subtilis
  JOURNAL   Nucleic Acids Res. 34 (9), E71 (2006)
  PUBMED    16714443
REFERENCE   2  (bases 1 to 8965)
  AUTHORS   Zhang X-Z., Yan X, Li S-P.
  TITLE     Direct Submission
  JOURNAL   Submitted (21-MAR-2006) Department of Microbiology, College of Life 
            Sciences, Nanjing Agricultural University, Key Laboratory for 
            Microbiological Engineering of Agricultural Environment, Ministry of
            Agriculture, 6 Tongwei Road, Nanjing, Jiangsu 210095, China
REFERENCE   3  (bases 1 to 8965)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8965
                     /organism="Delivery vector pIEFBPR"
                     /lab_host="Bacillus subtilis"
                     /mol_type="other DNA"
                     /db_xref="taxon:380061"
     rep_origin      4..459
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    692..1697
                     /note="bpr-front; upstream of Bacillus subtilis bpr gene; 
                     homologous arm for recombination"
     repeat_region   1728..1852
                     /label=DR sequence
                     /note="DR sequence"
     CDS             complement(1900..2982)
                     /codon_start=1
                     /gene="lacI"
                     /product="LacI"
                     /label=lacI
                     /note="lac operon repressor from Escherichia coli; 
                     engineered for expression in Gram-positive bacteria"
                     /protein_id="ABF31008.1"
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPSTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     gene            complement(1900..2982)
                     /gene="lacI"
                     /label=lacI
     CDS             complement(3232..3567)
                     /codon_start=1
                     /gene="mazF"
                     /product="MazF"
                     /label=mazF
                     /note="Escherichia coli toxin MazF"
                     /protein_id="ABF31009.1"
                     /translation="MVSRYVPDMGDLIWVDFDPTKGSEQAGHRPAVVLSPFMYNNKTGM
                     CLCVPCTTQSKGYPFEVVLSGQERDGVALADQVKSIAWRARGATKKGTVAPEELQLIKA
                     KINVLIG"
     gene            complement(3232..3567)
                     /gene="mazF"
                     /label=mazF
     protein_bind    3591..3607
                     /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)."
     CDS             4179..4931
                     /codon_start=1
                     /gene="spc"
                     /product="Spc"
                     /label=spc
                     /note="spectinomycin adenyltransferase; selectable either 
                     in Escherichia coli or Bacillus subtilis"
                     /protein_id="ABF31010.1"
                     /translation="MNTYEQINKVKKILRKHLKNNLIGTYMFGSGVESGLKPNSDLDFL
                     VVVSEPLTDQSKEILIQKIRPISKKIGDKSNLRYIELTIIIQQEMVPWNHPPKQEFIYG
                     EWLQELYEQGYIPQKELNSDLTIMLYQAKRKNKRIYGNYDLEELLPDIPFSDVRRAIMD
                     SSEELIDNYQDDETNSILTLCRMILTMDTGKIIPKDIAGNAVAESSPLEHRERILLAVR
                     SYLGENIEWTNENVNLTINYLNNRLKKL"
     gene            4179..4931
                     /gene="spc"
                     /label=spc
     repeat_region   4933..5057
                     /label=DR sequence
                     /note="DR sequence"
     misc_feature    5197..6235
                     /note="bpr-back; downstream of Bacillus subtilis bpr gene; 
                     homologous arm for recombination"
     CDS             6510..6758
                     /codon_start=1
                     /gene="mazE"
                     /product="MazE"
                     /label=mazE
                     /note="Escherichia coli anti-toxin MazE; counteracts toxin 
                     MazF"
                     /protein_id="ABF31011.1"
                     /translation="MIHSSVKRWGNSPAVRIPATLMQALNLNIDDEVKIDLVDGKLIIE
                     PVRKEPVFTLAELVNDITPENLHENIDWGEPKDKEVW"
     gene            6510..6758
                     /gene="mazE"
                     /label=mazE
     promoter        complement(6777..6795)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6816..6832)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    6840..6856
                     /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)."
     promoter        complement(6864..6894)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6909..6930
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(7035..7826)
                     /direction=LEFT
                     /label=ColE1 ori for Escherichia coli
                     /note="ColE1 ori for Escherichia coli"
     rep_origin      complement(7218..7806)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7977..8837)
                     /codon_start=1
                     /gene="bla"
                     /product="Bla"
                     /label=bla
                     /note="beta-lactamase; selectable in Escherichia coli only"
                     /protein_id="ABF31012.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(7977..8837)
                     /gene="bla"
                     /label=bla
     promoter        complement(8838..8942)
                     /gene="bla"
                     /label=AmpR promoter