LOCUS       Exported                5799 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Bicistronic shuttle vector pDUO-28T, complete sequence.
ACCESSION   KX806600
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5799)
  AUTHORS   Nakata PA.
  TITLE     Construction of pDUO: A bicistronic shuttle vector series for dual 
            expression of recombinant proteins
  JOURNAL   Plasmid 89, 16-21 (2016)
  PUBMED    27989736
REFERENCE   2  (bases 1 to 5799)
  AUTHORS   Nakata PA.
  TITLE     Direct Submission
  JOURNAL   Submitted (31-AUG-2016) Pediatrics, USDA/ARS Childrens Nutrition 
            Research Center, Baylor College of Medicine, 1100 Bates Street, 
            Houston, TX 77030-2600, USA
REFERENCE   3  (bases 1 to 5799)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..5799
                     /organism="Bicistronic shuttle vector pDUO-28T"
                     /mol_type="other DNA"
                     /db_xref="taxon:1897404"
     CDS             complement(657..893)
                     /codon_start=1
                     /gene="dhfrII"
                     /product="DhfRII"
                     /label=dhfrII
                     /note="dihydrofolate reductase"
                     /protein_id="APR64516.1"
                     /translation="MGQSSDEANAPVAGQFALPLSATFGLGDRVRKKSGAAWQGQVVGW
                     YCTKLTPEGYAVESESHPGSVQIYPVAALERVA"
     gene            complement(657..893)
                     /gene="dhfrII"
                     /label=dhfrII
     promoter        complement(1220..1248)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     rep_origin      1262..1662
                     /label=ori_pRO1600
                     /note="ori_pRO1600"
     rep_origin      1298..1649
                     /label=pRO1600 oriV
                     /note="broad-host-range origin of replication from 
                     Pseudomonas aeruginosa plasmid pRO1600; requires the 
                     pRO1600 Rep protein for replication (West et al., 1994)"
     CDS             1663..2496
                     /codon_start=1
                     /gene="rep"
                     /product="Rep"
                     /label=rep
                     /note="replication protein"
                     /protein_id="APR64517.1"
                     /translation="MASPPMVYKSNALVEAAYRLSVQEQRIVLACISQVKRSEPVTDEV
                     MYSVTAEDIATMAGVPIESSYNQLKEAALRLKRREVRLTQEPNGKGKRPSVMITGWVQT
                     IIYREGEGRVELRFTKDMLPYLTELTKQFTKYALADVAKMDSTHAIRLYELLMQWDSIG
                     QREIEIDQLRKWFQLEGRYPSIKDFKLRVLDPAVTQINEHSPLQVEWAQRKTGRKVTHL
                     LFSFGPKKPAKAVGKAPAKRKAGKISDAEIAKQARPGETWEAARARLTQMPLDLA"
     gene            1663..2496
                     /gene="rep"
                     /label=rep
     terminator      2522..2568
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     regulatory      2531..2688
                     /regulatory_class="terminator"
                     /note="rrnB T1 and T2 trancription terminator"
     misc_feature    2767..2811
                     /label=S tag
                     /note="S tag"
     CDS             complement(2767..2811)
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     misc_feature    complement(2818..2876)
                     /label=multiple cloning site
                     /note="multiple cloning site"
     RBS             2894..2916
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     misc_feature    2894..2899
                     /label=RBS
                     /note="RBS"
     promoter        complement(2943..3227)
                     /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)"
     misc_feature    2977..3015
                     /label=I2 and I1 region
                     /note="I2 and I1 region"
     misc_feature    3012..3025
                     /label=CAP binding site
                     /note="CAP binding site"
     misc_feature    3046..3067
                     /label=O1 region
                     /note="O1 region"
     misc_feature    3209..3224
                     /label=O2 region
                     /note="O2 region"
     terminator      3233..3279
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     regulatory      3242..3399
                     /regulatory_class="terminator"
                     /note="rrnB T1 and T2 trancription terminator"
     misc_feature    complement(3469..3554)
                     /label=multiple cloning site
                     /note="multiple cloning site"
     misc_feature    3524..3541
                     /label=HIS tag
                     /note="HIS tag"
     CDS             complement(3524..3541)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             3565..3587
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     misc_feature    3565..3570
                     /label=RBS
                     /note="RBS"
     promoter        complement(3614..3898)
                     /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)"
     misc_feature    3648..3686
                     /label=I2 and I1 region
                     /note="I2 and I1 region"
     misc_feature    3683..3696
                     /label=CAP binding site
                     /note="CAP binding site"
     misc_feature    3717..3738
                     /label=O1 region
                     /note="O1 region"
     misc_feature    3880..3895
                     /label=O2 region
                     /note="O2 region"
     CDS             3925..4803
                     /codon_start=1
                     /gene="araC"
                     /product="AraC"
                     /label=araC
                     /note="arabinose operon regulator"
                     /protein_id="APR64518.1"
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     gene            3925..4803
                     /gene="araC"
                     /label=araC
     oriT            4808..5075
     oriT            4960..5068
                     /note="incP origin of transfer"
     rep_origin      5125..5744
                     /label=ori_pBR322
                     /note="ori_pBR322"
     rep_origin      complement(5140..5728)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"