LOCUS       Exported                3677 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pCDF-J23114-TetO-lacI-LVA, complete sequence.
ACCESSION   KM229382
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3677)
  AUTHORS   Schaerli Y, Munteanu A, Gili M, Cotterell J, Sharpe J, Isalan M.
  TITLE     A unified design space of synthetic stripe-forming networks
  JOURNAL   Nat Commun 5, 4905 (2014)
  PUBMED    25247316
REFERENCE   2  (bases 1 to 3677)
  AUTHORS   Schaerli Y, Munteanu A, Gili M, Cotterell J, Sharpe J, Isalan M.
  TITLE     Direct Submission
  JOURNAL   Submitted (24-JUL-2014) Centre for Genomic Regulation (CRG), 
            EMBL/CRG Systems Biology Research Unit, Dr. Aiguader 88, Barcelona, 
            Barcelona 08003, Spain
REFERENCE   3  (bases 1 to 3677)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..3677
                     /organism="Cloning vector pCDF-J23114-TetO-lacI-LVA"
                     /mol_type="other DNA"
                     /db_xref="taxon:1531941"
     CDS             complement(114..905)
                     /codon_start=1
                     /gene="specR"
                     /product="spectinomycin resistance protein"
                     /label=specR
                     /note="confers resistance to spectinomycin"
                     /protein_id="AIO11014.1"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     gene            complement(114..905)
                     /gene="specR"
                     /label=specR
     promoter        complement(906..997)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1045..1783)
                     /direction=LEFT
                     /label=CloDF13 ori
                     /note="Plasmids containing the CloDF13 (CDF) origin of 
                     replication can be propagated in E. coli cells that contain
                     additional plasmids with compatible origins."
     primer_bind     1946..1962
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     gene            1975..3293
                     /gene="lacI-LVA"
                     /label=lacI-LVA
     regulatory      1975..2009
                     /regulatory_class="promoter"
                     /gene="lacI-LVA"
                     /note="J23114 promoter"
     misc_binding    2016..2034
                     /gene="lacI-LVA"
                     /label=TetR-LVA binding site
                     /bound_moiety="TetR-LVA"
                     /note="TetO"
     protein_bind    2016..2034
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             2142..2164
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2172..3293
                     /codon_start=1
                     /gene="lacI-LVA"
                     /product="lac repressor"
                     /label=lacI-LVA
                     /note="DNA-binding protein with degradation tag (LVA)"
                     /protein_id="AIO11013.1"
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQRPAANDENYALVA"
     CDS             2172..3251
                     /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"
     regulatory      3401..3653
                     /regulatory_class="terminator"
     terminator      3401..3448
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      3457..3543
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3635..3662
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"