LOCUS       40924_16305        5799 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Bicistronic shuttle vector pDUO-28T, complete sequence.
ACCESSION   .
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 Children's Nutrition 
            Research Center, Baylor College of Medicine, 1100 Bates Street, 
            Houston, TX 77030-2600, USA
REFERENCE   3  (bases 1 to 5799)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5799)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Plasmid 89,
            16-21 (2016)"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (31-AUG-2016) Pediatrics, USDA/ARS Children's Nutrition Research 
            Center, Baylor College of Medicine, 1100 Bates Street, Houston, TX 
            77030-2600, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..5799
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(660..893)
                     /label=TpR
                     /note="E. coli plasmid-associated dihydrofolate reductase"
     promoter        complement(1220..1248)
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     rep_origin      complement(1298..1649)
                     /direction=LEFT
                     /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..2493
                     /label=pRO1600 Rep
                     /note="replication protein for the broad-host-range plasmid
                     pRO1600 from Pseudomonas aeruginosa"
     terminator      2522..2565
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     CDS             complement(2767..2811)
                     /label=S-Tag
                     /note="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
     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      complement(3233..3279)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     CDS             complement(3524..3541)
                     /label=6xHis
                     /note="6xHis affinity tag"
     RBS             complement(3565..3587)
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     promoter        complement(3614..3898)
                     /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)"
     CDS             3925..4800
                     /label=araC
                     /note="L-arabinose regulatory protein"
     oriT            4960..5068
                     /label=oriT
                     /note="incP origin of transfer"
     rep_origin      complement(5140..5728)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
       61 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
      121 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
      181 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
      241 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
      301 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
      361 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
      421 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
      481 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
      541 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
      601 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtttcttag
      661 gccacacgtt caagtgcagc cacaggataa atttgcactg agcctgggtg ggattcggac
      721 tcgaccgcat agccttcagg agtgagtttt gtgcaatacc aaccgacgac ttgaccctgc
      781 caagcggcac cagatttctt gcgtacgcga tcccctaagc caaaggtggc actcagggga
      841 agcgcaaact gccctgcaac gggagcgttg gcttcatcgc tactttgacc catgtcgaat
      901 ccttcttgtg aatctattat ggcgacaagc aaatcgaact ctgactgcct accccacaac
      961 aactactaga aagcaccagc acaacggctg cctaactttg ttttagggcg actaccctgc
     1021 tgcgtaacat cgttgctgtt ccataacatc aaacatcgac ccacggcgta acgcgcttag
     1081 ctagcttgga tgcccgaggc tagactgtac aaaaaaacag tcataacaag ccatgaaaac
     1141 cgccactgcg ccgttaccac cgctgcgttc ggtcaaggtt ctggaccagt tgcgtgagcg
     1201 catacgctac ttgcattaca gtttacgaac cgaacaggct tatgtcaact gggttcgtga
     1261 acgccgctgg tgccgctggt tggacgccaa gggtgaatcc gcctcgatac cctgattact
     1321 cgcttcctgc gccctctcag gcggcgatag gggactggta aaacggggat tgcccagacg
     1381 cctcccccgc cccttcaggg gcacaaatgc ggccccaacg gggccacgta gtggtgcgtt
     1441 ttttgcgttt ccaccctttt cttccttttc ccttttaaac cttttaggac gtctacaggc
     1501 cacgtaatcc gtggcctgta gagtttaaaa agggacggat ttgttgccat taagggacgg
     1561 atttgttgtt aagaagggac ggatttgttg ttgtaaaggg acggatttgt tgtattgtgg
     1621 gacgcagata cagtgtcccc ttatacacaa ggaatgtcga acgtggcctc acccccaatg
     1681 gtttacaaaa gcaatgccct ggtcgaggcc gcgtatcgcc tcagtgttca ggaacagcgg
     1741 atcgttctgg cctgtattag ccaggtgaag aggagcgagc ctgtcaccga tgaagtgatg
     1801 tattcagtga cggcggagga catagcgacg atggcgggtg tccctatcga atcttcctac
     1861 aaccagctca aagaagcggc cctgcgcctg aaacggcggg aagtccggtt aacccaagag
     1921 cccaatggca aggggaaaag accgagtgtg atgattaccg gctgggtgca aacaatcatc
     1981 taccgggagg gtgagggccg tgtagaactc aggttcacca aagacatgct gccgtacctg
     2041 acggaactca ccaaacagtt caccaaatac gccttggctg acgtggccaa gatggacagc
     2101 acccacgcga tcaggcttta cgagctgctc atgcaatggg acagcatcgg ccagcgcgaa
     2161 atagaaattg accagctgcg aaagtggttt caactggaag gccggtatcc ctcgatcaag
     2221 gacttcaagt tgcgagtgct tgatccagcc gtgacgcaga tcaacgagca cagcccgcta
     2281 caggtggagt gggcgcagcg aaagaccggg cgcaaggtca cacatctgtt gttcagtttt
     2341 ggaccgaaga agcccgccaa ggcggtgggt aaggccccag cgaagcgcaa ggccgggaag
     2401 atttcagatg ctgagatcgc gaaacaggct cgccctggtg agacatggga agcggcccgc
     2461 gctcgactaa cccagatgcc gctggatctg gcctagaggc cgtggccacc acggccaaca
     2521 gataaaacga aaggcccagt ctttcgactg agcctttcgt tttatttgat gcctggcagt
     2581 tccctactct cgcatgggga gaccccacac taccatcggc gctacggcgt ttcacttctg
     2641 agttcggcat ggggtcaggt gggaccaccg cgctactgcc gccaggcaaa ttctgtttta
     2701 tcagaccgct tctgcgttct gatttaatct gtatcaggct gaggttaatt aagctgcgct
     2761 agtagacgag tccatgtgct ggcgttcaaa tttcgcagca gcggtttctt taccagactc
     2821 gagggtaccg acgtcagcga tcgcgtggcc ggccgatatc caattgagat ctgccatatg
     2881 tatagtatgt atatctcctt cttaaagtta aacaaaatta tttctagccc aaaaaaacgg
     2941 gtatggagaa acagtagaga gttgcgataa aaagcgtcag gtaggatccg ctaatcttat
     3001 ggataaaaat gctatggcat agcaaagtgt gacgccgtgc aaataatcaa tgtggacttt
     3061 tctgccgtga ttatagacac ttttgttacg cgtttttgtc atggctttgg tcccgctttg
     3121 ttacagaatg cttttaataa gcggggttac cggtttggtt agcgagaaga gccagtaaaa
     3181 gacgcagtga cggcaatgtc tgatgcaata tggacaattg gtttcttaac agataaaacg
     3241 aaaggcccag tctttcgact gagcctttcg ttttatttga tgcctggcag ttccctactc
     3301 tcgcatgggg agaccccaca ctaccatcgg cgctacggcg tttcacttct gagttcggca
     3361 tggggtcagg tgggaccacc gcgctactgc cgccaggcaa attctgtttt atttcgatta
     3421 tgcggccgtg tacaatacga ttactttctg ttcgacttaa gcattatgcg gccgcaagct
     3481 tgtcgacctg caggcgcgcc gagctcgaat tcggatcctg gctgtggtga tgatggtgat
     3541 ggctgctgcc catgggtatg tatatctcct tcttaaagtt aaacaaaatt atttctagcc
     3601 caaaaaaacg ggtatggaga aacagtagag agttgcgata aaaagcgtca ggtaggatcc
     3661 gctaatctta tggataaaaa tgctatggca tagcaaagtg tgacgccgtg caaataatca
     3721 atgtggactt ttctgccgtg attatagaca cttttgttac gcgtttttgt catggctttg
     3781 gtcccgcttt gttacagaat gcttttaata agcggggtta ccggtttggt tagcgagaag
     3841 agccagtaaa agacgcagtg acggcaatgt ctgatgcaat atggacaatt ggtttcttct
     3901 ctgaatggcg ggagtatgaa aagtatggct gaagcgcaaa atgatcccct gctgccggga
     3961 tactcgttta atgcccatct ggtggcgggt ttaacgccga ttgaggccaa cggttatctc
     4021 gattttttta tcgaccgacc gctgggaatg aaaggttata ttctcaatct caccattcgc
     4081 ggtcaggggg tggtgaaaaa tcagggacga gaatttgttt gccgaccggg tgatattttg
     4141 ctgttcccgc caggagagat tcatcactac ggtcgtcatc cggaggctcg cgaatggtat
     4201 caccagtggg tttactttcg tccgcgcgcc tactggcatg aatggcttaa ctggccgtca
     4261 atatttgcca atacggggtt ctttcgcccg gatgaagcgc accagccgca tttcagcgac
     4321 ctgtttgggc aaatcattaa cgccgggcaa ggggaagggc gctattcgga gctgctggcg
     4381 ataaatctgc ttgagcaatt gttactgcgg cgcatggaag cgattaacga gtcgctccat
     4441 ccaccgatgg ataatcgggt acgcgaggct tgtcagtaca tcagcgatca cctggcagac
     4501 agcaattttg atatcgccag cgtcgcacag catgtttgct tgtcgccgtc gcgtctgtca
     4561 catcttttcc gccagcagtt agggattagc gtcttaagct ggcgcgagga ccaacgtatc
     4621 agccaggcga agctgctttt gagcaccacc cggatgccta tcgccaccgt cggtcgcaat
     4681 gttggttttg acgatcaact ctatttctcg cgggtattta aaaaatgcac cggggccagc
     4741 ccgagcgagt tccgtgccgg ttgtgaagaa aaagtgaatg atgtagccgt caagttgtca
     4801 taatttgggg attccttaag gtatactttc cgctgcataa ccctgcttcg gggtcattat
     4861 agcgattttt tcggtatatc catccttttt cgcacgatat acaggatttt gccaaagggt
     4921 tcgtgtagac tttccttggt gtatccaacg gcgtcagccg ggcaggatag gtgaagtagg
     4981 cccacccgcg agcgggtgtt ccttcttcac tgtcccttat tcgcacctgg cggtgctcaa
     5041 cgggaatcct gctctgcgag gctggccgat aagctagctt atgtgagcaa aaggccagca
     5101 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     5161 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     5221 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     5281 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     5341 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     5401 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     5461 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     5521 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     5581 gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     5641 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     5701 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     5761 cgctcagtgg aacgaaaact cacgttaagg gattttggt
//
