LOCUS       Exported                8619 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pVWEx1, complete sequence.
ACCESSION   MF034723
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8619)
  AUTHORS   Bekel T, Stadermann KB, Voss M.
  TITLE     Nucleotide sequence analysis of the plasmid pVWEx1 by Illumina 
            sequencing technology
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8619)
  AUTHORS   Bekel T, Stadermann KB, Voss M.
  TITLE     Direct Submission
  JOURNAL   Submitted (28-APR-2017) Evonik Nutrition & Care GmbH, Kantstrasse 2,
            Halle 33790, Germany
REFERENCE   3  (bases 1 to 8619)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Assembly Method       :: CLC Genomics Workbench v. 9
            Sequencing Technology :: Illumina
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..8619
                     /organism="Cloning vector pVWEx1"
                     /mol_type="other DNA"
                     /label=derived from Corynebacterium glutamicum
                     /note="derived from Corynebacterium glutamicum"
                     /db_xref="taxon:2005483"
     regulatory      1..80
                     /regulatory_class="promoter"
                     /note="tacI"
     promoter        7..35
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    43..59
                     /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)."
     misc_feature    105..161
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     misc_feature    117..140
                     /label=multiple cloning site
                     /note="multiple cloning site"
     primer_bind     complement(162..178)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      391..477
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      569..596
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        615..706
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      924..3989
                     /label=derived from Corynebacterium glutamicum pCG1
                     /note="derived from Corynebacterium glutamicum pCG1"
     CDS             complement(4572..5387)
                     /codon_start=1
                     /gene="neo"
                     /locus_tag="pVWEx1_neo"
                     /product="kanamycin kinase"
                     /label=neo
                     /note="aminoglycoside 3-phosphotransferase"
                     /protein_id="ARU80374.1"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     gene            complement(4572..5387)
                     /gene="neo"
                     /locus_tag="pVWEx1_neo"
                     /label=neo
     rep_origin      complement(5715..6544)
                     /direction=LEFT
                     /label=derived from Escherichia coli p15A
                     /note="derived from Escherichia coli p15A"
     rep_origin      complement(5982..6527)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin."
     CDS             complement(7232..8323)
                     /codon_start=1
                     /gene="lacI"
                     /locus_tag="pVWEx1_lacI"
                     /product="lactose operon repressor"
                     /label=lacI
                     /note="LacI"
                     /protein_id="ARU80375.1"
                     /translation="MVNVKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAM
                     AELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVER
                     SGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSII
                     FSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGD
                     WSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTED
                     SSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTA
                     SPRALADSLMQLARQVSRLESGQ"
     gene            complement(7232..8323)
                     /gene="lacI"
                     /locus_tag="pVWEx1_lacI"
                     /label=lacI
     CDS             complement(7232..8314)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     promoter        complement(8315..8392)
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."