LOCUS       Exported                6022 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Bicistronic shuttle vector pDUO-30T, complete sequence.
ACCESSION   KX806601
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6022)
  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 6022)
  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 6022)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..6022
                     /organism="Bicistronic shuttle vector pDUO-30T"
                     /mol_type="other DNA"
                     /db_xref="taxon:1897405"
     CDS             complement(730..1263)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyl transferase"
                     /label=aacC1
                     /note="gentamycin resistance marker"
                     /protein_id="APR64519.1"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     gene            complement(730..1263)
                     /gene="aacC1"
                     /label=aacC1
     promoter        complement(1452..1480)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     rep_origin      1485..1885
                     /label=ori_pRO1600
                     /note="ori_pRO1600"
     rep_origin      1521..1872
                     /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             1886..2719
                     /codon_start=1
                     /gene="rep"
                     /product="Rep"
                     /label=rep
                     /note="replication protein"
                     /protein_id="APR64520.1"
                     /translation="MASPPMVYKSNALVEAAYRLSVQEQRIVLACISQVKRSEPVTDEV
                     MYSVTAEDIATMAGVPIESSYNQLKEAALRLKRREVRLTQEPNGKGKRPSVMITGWVQT
                     IIYREGEGRVELRFTKDMLPYLTELTKQFTKYALADVAKMDSTHAIRLYELLMQWDSIG
                     QREIEIDQLRKWFQLEGRYPSIKDFKLRVLDPAVTQINEHSPLQVEWAQRKTGRKVTHL
                     LFSFGPKKPAKAVGKAPAKRKAGKISDAEIAKQARPGETWEAARARLTQMPLDLA"
     gene            1886..2719
                     /gene="rep"
                     /label=rep
     terminator      2745..2791
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     regulatory      2754..2911
                     /regulatory_class="terminator"
                     /note="rrnB T1 and T2 trancription terminator"
     misc_feature    2990..3034
                     /label=S tag
                     /note="S tag"
     CDS             complement(2990..3034)
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     misc_feature    complement(3041..3099)
                     /label=multiple cloning site
                     /note="multiple cloning site"
     RBS             3117..3139
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     misc_feature    3117..3122
                     /label=RBS
                     /note="RBS"
     promoter        complement(3166..3450)
                     /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    3200..3238
                     /label=I2 and I1 region
                     /note="I2 and I1 region"
     misc_feature    3235..3248
                     /label=CAP binding site
                     /note="CAP binding site"
     misc_feature    3269..3290
                     /label=O1 region
                     /note="O1 region"
     misc_feature    3432..3447
                     /label=O2 region
                     /note="O2 region"
     terminator      3456..3502
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     regulatory      3465..3622
                     /regulatory_class="terminator"
                     /note="rrnB T1 and T2 trancription terminator"
     misc_feature    complement(3692..3774)
                     /label=multiple cloning site
                     /note="multiple cloning site"
     misc_feature    3747..3764
                     /label=HIS tag
                     /note="HIS tag"
     CDS             complement(3747..3764)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             3788..3810
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     misc_feature    3788..3793
                     /label=RBS
                     /note="RBS"
     promoter        complement(3837..4121)
                     /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    3871..3909
                     /label=I2 and I1 region
                     /note="I2 and I1 region"
     misc_feature    3906..3919
                     /label=CAP binding site
                     /note="CAP binding site"
     misc_feature    3940..3961
                     /label=O1 region
                     /note="O1 region"
     misc_feature    4103..4118
                     /label=O2 region
                     /note="O2 region"
     CDS             4148..5026
                     /codon_start=1
                     /gene="araC"
                     /product="AraC"
                     /label=araC
                     /note="arabinose operon regulator"
                     /protein_id="APR64521.1"
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     gene            4148..5026
                     /gene="araC"
                     /label=araC
     oriT            5030..5298
     oriT            5183..5291
                     /note="incP origin of transfer"
     rep_origin      5348..5967
                     /label=ori_pBR322
                     /note="ori_pBR322"
     rep_origin      complement(5363..5951)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"