LOCUS       Exported                8463 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Bacterial expression vector pJBtrc10, complete sequence.
ACCESSION   LT727349
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8463)
  AUTHORS   De Schamphelaire W, Olbrechts A, Meert J, Verhelst K, Roggeman 
            Fonseca M, Vanhoucke M, Beyaert R.
  TITLE     BCCM/LMBP Plasmid collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8463)
  AUTHORS   De Schamphelaire W.
  TITLE     Direct Submission
  JOURNAL   Submitted (02-FEB-2017) BCCM/LMBP, Universiteit Gent, 
            Technologiepark 927, 9052, BELGIUM
REFERENCE   3  (bases 1 to 8463)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8463
                     /organism="Bacterial expression vector pJBtrc10"
                     /mol_type="other DNA"
                     /note="BCCM/LMBP Plasmid collection (Ghent 
                     University,Belgium) accession number LMBP 3523."
                     /db_xref="taxon:1944682"
     CDS             complement(793..1983)
                     /codon_start=1
                     /note="unnamed protein product; tetracycline resistance"
                     /protein_id="SJL88180.1"
                     /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
                     GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
                     VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
                     LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
                     QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
                     AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
                     AALTSLTSITGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
     promoter        complement(2031..2059)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(2072..2088)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(2696..3844)
                     /codon_start=1
                     /note="unnamed protein product; trfA"
                     /protein_id="SJL88181.1"
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     MVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     regulatory      complement(3953..4002)
                     /regulatory_class="promoter"
                     /note="Tn5 neomycin resistance promoter"
     rep_origin      4236..4766
                     /label=oriV
                     /note="incP origin of replication"
     rep_origin      complement(4242..4766)
                     /direction=LEFT
                     /label=RK2 ori
                     /note="RK2 ori"
     regulatory      5249..5326
                     /regulatory_class="promoter"
                     /note="lacIq promoter"
     promoter        5249..5326
                     /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."
     CDS             5327..6409
                     /codon_start=1
                     /note="unnamed protein product; lacI"
                     /protein_id="SJL88182.1"
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPSTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     regulatory      6632..6675
                     /regulatory_class="promoter"
                     /note="trc promoter"
     promoter        6641..6670
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     misc_feature    6676..6700
                     /label=5 UTR of lacZ
                     /note="5 UTR of lacZ"
     protein_bind    6678..6694
                     /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    6701..6742
                     /label=5 UTR of T7 gene 10, incl. RBS
                     /note="5 UTR of T7 gene 10, incl. RBS"
     RBS             6713..6735
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     promoter        complement(6797..6814)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     regulatory      complement(6799..6815)
                     /regulatory_class="promoter"
                     /note="T3 promoter"
     regulatory      6833..6880
                     /regulatory_class="terminator"
                     /note="T7 terminator"
     terminator      6833..6880
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     regulatory      6954..7001
                     /regulatory_class="terminator"
                     /note="T7 terminator"
     terminator      6954..7001
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     regulatory      7070..7151
                     /regulatory_class="terminator"
                     /note="fd terminator"
     terminator      7073..7121
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     regulatory      7180..7261
                     /regulatory_class="terminator"
                     /note="fd terminator"
     terminator      7183..7231
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     promoter        complement(7897..8001)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(8176..8286)
                     /direction=LEFT
                     /label=RK2/RP4 oriT
                     /note="RK2/RP4 oriT"
     oriT            8178..8286
                     /note="incP origin of transfer"
     primer_bind     complement(8373..8389)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"