LOCUS       Exported                5893 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Bicistronic shuttle vector pDUO-20T, complete sequence.
ACCESSION   KX806598
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5893)
  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 5893)
  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 5893)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..5893
                     /organism="Bicistronic shuttle vector pDUO-20T"
                     /mol_type="other DNA"
                     /db_xref="taxon:1897402"
     promoter        96..200
                     /gene="bla"
                     /label=AmpR promoter
     CDS             201..1061
                     /codon_start=1
                     /gene="bla"
                     /product="Bla"
                     /label=bla
                     /note="beta-lactamase"
                     /protein_id="APR64510.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            201..1061
                     /gene="bla"
                     /label=bla
     rep_origin      1216..1835
                     /label=ori_pBR322
                     /note="ori_pBR322"
     rep_origin      1232..1820
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     oriT            1885..2152
     oriT            1892..2000
                     /note="incP origin of transfer"
     CDS             complement(2157..3035)
                     /codon_start=1
                     /gene="araC"
                     /product="AraC"
                     /label=araC
                     /note="arabinose operon regulator"
                     /protein_id="APR64511.1"
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     gene            complement(2157..3035)
                     /gene="araC"
                     /label=araC
     promoter        3062..3346
                     /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    3065..3080
                     /label=O2 region
                     /note="O2 region"
     misc_feature    3222..3243
                     /label=O1 region
                     /note="O1 region"
     misc_feature    3264..3277
                     /label=CAP binding site
                     /note="CAP binding site"
     misc_feature    3274..3312
                     /label=I2 and I1 region
                     /note="I2 and I1 region"
     RBS             3373..3395
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     misc_feature    3390..3395
                     /label=RBS
                     /note="RBS"
     misc_feature    3406..3487
                     /label=multiple cloning site
                     /note="multiple cloning site"
     misc_feature    3419..3436
                     /label=HIS tag
                     /note="HIS tag"
     CDS             3419..3436
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     regulatory      3561..3718
                     /regulatory_class="terminator"
                     /note="rrnB T1 and T2 trancription terminator"
     terminator      3681..3727
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     promoter        3733..4017
                     /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    3736..3751
                     /label=O2 region
                     /note="O2 region"
     misc_feature    3893..3914
                     /label=O1 region
                     /note="O1 region"
     misc_feature    3935..3948
                     /label=CAP binding site
                     /note="CAP binding site"
     misc_feature    3945..3983
                     /label=I2 and I1 region
                     /note="I2 and I1 region"
     RBS             4044..4066
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     misc_feature    4061..4066
                     /label=RBS
                     /note="RBS"
     misc_feature    4081..4137
                     /label=multiple cloning site
                     /note="multiple cloning site"
     misc_feature    4149..4193
                     /label=S tag
                     /note="S tag"
     CDS             4149..4193
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     regulatory      4272..4429
                     /regulatory_class="terminator"
                     /note="rrnB T1 and T2 trancription terminator"
     terminator      4392..4438
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(4464..5297)
                     /codon_start=1
                     /gene="rep"
                     /product="Rep"
                     /label=rep
                     /note="replication protein"
                     /protein_id="APR64512.1"
                     /translation="MASPPMVYKSNALVEAAYRLSVQEQRIVLACISQVKRSEPVTDEV
                     MYSVTAEDIATMAGVPIESSYNQLKEAALRLKRREVRLTQEPNGKGKRPSVMITGWVQT
                     IIYREGEGRVELRFTKDMLPYLTELTKQFTKYALADVAKMDSTHAIRLYELLMQWDSIG
                     QREIEIDQLRKWFQLEGRYPSIKDFKLRVLDPAVTQINEHSPLQVEWAQRKTGRKVTHL
                     LFSFGPKKPAKAVGKAPAKRKAGKISDAEIAKQARPGETWEAARARLTQMPLDLA"
     gene            complement(4464..5297)
                     /gene="rep"
                     /label=rep
     rep_origin      5298..5707
                     /label=ori_pRO1600
                     /note="ori_pRO1600"
     rep_origin      5311..5662
                     /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)"