LOCUS       Exported                6055 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pJN105, complete sequence.
ACCESSION   AF179381
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6055)
  AUTHORS   Newman JR, Fuqua C.
  TITLE     Broad-host-range expression vectors that carry the 
            L-arabinose-inducible Escherichia coli araBAD promoter and the araC 
            regulator
  JOURNAL   Gene 227 (2), 197-203 (1999)
  PUBMED    10023058
REFERENCE   2  (bases 1 to 6055)
  AUTHORS   Newman JR, Fuqua C.
  TITLE     Direct Submission
  JOURNAL   Submitted (21-AUG-1999) Biology, Indiana University, JH142, 1001 E. 
            3rd St., Bloomington, IN 47405-6801, USA
REFERENCE   3  (bases 1 to 6055)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6055
                     /organism="Expression vector pJN105"
                     /mol_type="genomic DNA"
                     /note="broad host range (pBBR replicon) vector; araC-PBAD 
                     expression cassette"
                     /db_xref="taxon:102682"
     CDS             complement(261..794)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamicin-3-acetyltransferase"
                     /function="confers gentamicin resistance"
                     /label=aacC1
                     /note="AacC1"
                     /protein_id="AAF00475.1"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     gene            complement(261..794)
                     /gene="aacC1"
                     /label=aacC1
     promoter        complement(983..1011)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     protein_bind    1281..1302
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        1317..1347
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1355..1371
                     /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)."
     primer_bind     1379..1395
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1416..1434
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     1464..1480
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     gene            complement(1572..2630)
                     /gene="araC"
                     /label=araC
     CDS             complement(1572..2450)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose responsive transcription factor"
                     /label=araC
                     /note="AraC"
                     /protein_id="AAF00474.1"
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        2477..2761
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     protein_bind    2478..2497
                     /label=AraC binding site
                     /bound_moiety="AraC"
                     /note="araO2"
     regulatory      complement(2595..2630)
                     /regulatory_class="promoter"
                     /gene="araC"
                     /note="araC promoter"
     protein_bind    2616..2641
                     /label=CRP binding site
                     /bound_moiety="CRP"
                     /note="PC region"
     protein_bind    2618..2657
                     /label=AraC binding site
                     /bound_moiety="AraC"
                     /note="araO1"
     protein_bind    2655..2684
                     /label=CRP binding site
                     /bound_moiety="CRP"
                     /note="PBAD region"
     protein_bind    2689..2726
                     /label=AraC binding site
                     /bound_moiety="AraC"
                     /note="araI"
     regulatory      2724..2762
                     /regulatory_class="promoter"
                     /note="araBAD promoter"
     misc_feature    2776..2841
                     /label=multiple cloning site
                     /note="multiple cloning site"
     primer_bind     complement(2801..2817)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(2850..2868)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2878..2894)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(3601..4263)
                     /codon_start=1
                     /gene="rep"
                     /product="replication protein"
                     /label=rep
                     /note="Rep; required for pBBR-type plasmid replication"
                     /protein_id="AAF00476.1"
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
     gene            complement(3601..4263)
                     /gene="rep"
                     /label=rep
     rep_origin      4264..5033
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"