LOCUS       Exported                8247 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Delivery vector pIEF16S, complete sequence.
ACCESSION   DQ358864
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8247)
  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 8247)
  AUTHORS   Zhang X-Z., Li S-P.
  TITLE     Direct Submission
  JOURNAL   Submitted (10-JAN-2006) Department of Microbiology, College of Life 
            Sciences, Nanjing Agricultral University, Key Laboratory for 
            Microbiological Engineering of Agricultural Environment of Ministry 
            of Agriculture, 6 Tongwei Road, Nanjing, Jiangsu 210095, China
REFERENCE   3  (bases 1 to 8247)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8247
                     /organism="Delivery vector pIEF16S"
                     /lab_host="Bacillus subtilis"
                     /mol_type="other DNA"
                     /db_xref="taxon:369433"
     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    complement(674..1067)
                     /note="downstream sequence of Bacillus subtilis 16S rRNA 
                     gene"
     repeat_region   1103..1227
     CDS             complement(1258..2340)
                     /codon_start=1
                     /gene="lacI"
                     /product="LacI"
                     /label=lacI
                     /note="lac repressor"
                     /protein_id="ABC88417.1"
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPSTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     gene            complement(1258..2340)
                     /gene="lacI"
                     /label=lacI
     CDS             complement(2590..2925)
                     /codon_start=1
                     /gene="mazF"
                     /product="MazF"
                     /label=mazF
                     /note="Escherichia coli toxin"
                     /protein_id="ABC88418.1"
                     /translation="MVSRYVPDMGDLIWVDFDPTKGSEQAGHRPAVVLSPFMYNNKTGM
                     CLCVPCTTQSKGYPFEVVLSGQERDGVALADQVKSIAWRARGATKKGTVAPEELQLIKA
                     KINVLIG"
     gene            complement(2590..2925)
                     /gene="mazF"
                     /label=mazF
     misc_feature    complement(2942..3230)
                     /label=Pspac promoter functional region
                     /note="Pspac promoter functional region"
     protein_bind    2949..2965
                     /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             3537..4289
                     /codon_start=1
                     /gene="spc"
                     /product="spectinomycin resistance marker"
                     /label=spc
                     /protein_id="ABC88419.1"
                     /translation="MNTYEQINKVKKILRKHLKNNLIGTYMFGSGVESGLKPNSDLDFL
                     VVVSEPLTDQSKEILIQKIRPISKKIGDKSNLRYIELTIIIQQEMVPWNHPPKQEFIYG
                     EWLQELYEQGYIPQKELNSDLTIMLYQAKRKNKRIYGNYDLEELLPDIPFSDVRRAIMD
                     SSEELIDNYQDDETNSILTLCRMILTMDTGKIIPKDIAGNAVAESSPLEHRERILLAVR
                     SYLGENIEWTNENVNLTINYLNNRLKKL"
     gene            3537..4289
                     /gene="spc"
                     /label=spc
     repeat_region   4291..4415
     misc_feature    complement(4538..5537)
                     /note="unstream sequence of Bacillus subtilis 16S rRNA 
                     gene"
     misc_feature    5544..5791
                     /note="promoter functional region of Escherichia coli lpp 
                     gene"
     CDS             5792..6040
                     /codon_start=1
                     /gene="mazE"
                     /product="MazE"
                     /label=mazE
                     /note="Escherichia coli anti-toxin"
                     /protein_id="ABC88420.1"
                     /translation="MIHSSVKRWGNSPAVRIPATLMQALNLNIDDEVKIDLVDGKLIIE
                     PVRKEPVFTLAELVNDITPENLHENIDWGEPKDKEVW"
     gene            5792..6040
                     /gene="mazE"
                     /label=mazE
     promoter        complement(6059..6077)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6098..6114)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    6122..6138
                     /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(6146..6176)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6191..6212
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      6317..7108
                     /label=ColE1
                     /note="ColE1"
     rep_origin      complement(6500..7088)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7259..8119)
                     /codon_start=1
                     /gene="amp"
                     /product="ampicillin resistance marker"
                     /label=amp
                     /protein_id="ABC88421.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(7259..8119)
                     /gene="amp"
                     /label=amp
     promoter        complement(8120..8224)
                     /gene="bla"
                     /label=AmpR promoter