LOCUS       40924_16310        6022 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Bicistronic shuttle vector pDUO-30T, complete sequence.
ACCESSION   .
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 Children's Nutrition 
            Research Center, Baylor College of Medicine, 1100 Bates Street, 
            Houston, TX 77030-2600, USA
REFERENCE   3  (bases 1 to 6022)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6022)
  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..6022
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(733..1263)
                     /label=GmR
                     /note="gentamycin acetyltransferase"
     promoter        complement(1452..1480)
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     rep_origin      complement(1521..1872)
                     /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             1886..2716
                     /label=pRO1600 Rep
                     /note="replication protein for the broad-host-range plasmid
                     pRO1600 from Pseudomonas aeruginosa"
     terminator      2745..2788
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     CDS             complement(2990..3034)
                     /label=S-Tag
                     /note="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
     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      complement(3456..3502)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     CDS             complement(3747..3764)
                     /label=6xHis
                     /note="6xHis affinity tag"
     RBS             complement(3788..3810)
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     promoter        complement(3837..4121)
                     /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             4148..5023
                     /label=araC
                     /note="L-arabinose regulatory protein"
     oriT            5183..5291
                     /label=oriT
                     /note="incP origin of transfer"
     rep_origin      complement(5363..5951)
                     /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 agtcgaattg
      661 gccgcggcgt tgtgacaatt taccgaacaa ctccgcggcc gggaagccga tctcggcttg
      721 aacgaattgt taggtggcgg tacttgggtc gatatcaaag tgcatcactt cttcccgtat
      781 gcccaacttt gtatagagag ccactgcggg atcgtcaccg taatctgctt gcacgtagat
      841 cacataagca ccaagcgcgt tggcctcatg cttgaggaga ttgatgagcg cggtggcaat
      901 gccctgcctc cggtgctcgc cggagactgc gagatcatag atatagatct cactacgcgg
      961 ctgctcaaac ctgggcagaa cgtaagccgc gagagcgcca acaaccgctt cttggtcgaa
     1021 ggcagcaagc gcgatgaatg tcttactacg gagcaagttc ccgaggtaat cggagtccgg
     1081 ctgatgttgg gagtaggtgg ctacgtctcc gaactcacga ccgaaaagat caagagcagc
     1141 ccgcatggat ttgacttggt cagggccgag cctacatgtg cgaatgatgc ccatacttga
     1201 gccacctaac tttgttttag ggcgactgcc ctgctgcgta acatcgttgc tgctgcgtaa
     1261 catcgttgct gctccataac atcaaacatc gacccacggc gtaacgcgct tgctgcttgg
     1321 atgcccgagg catagactgt acaaaaaaac agtcataaca agccatgaaa accgccactg
     1381 cgccgttacc accgctgcgt tcggtcaagg ttctggacca gttgcgtgag cgcatacgct
     1441 acttgcatta cagtttacga accgaacagg cttatgtcaa ttcgcgccgc tggtgccgct
     1501 ggttggacgc caagggtgaa tccgcctcga taccctgatt actcgcttcc tgcgccctct
     1561 caggcggcga taggggactg gtaaaacggg gattgcccag acgcctcccc cgccccttca
     1621 ggggcacaaa tgcggcccca acggggccac gtagtggtgc gttttttgcg tttccaccct
     1681 tttcttcctt ttccctttta aaccttttag gacgtctaca ggccacgtaa tccgtggcct
     1741 gtagagttta aaaagggacg gatttgttgc cattaaggga cggatttgtt gttaagaagg
     1801 gacggatttg ttgttgtaaa gggacggatt tgttgtattg tgggacgcag atacagtgtc
     1861 cccttataca caaggaatgt cgaacgtggc ctcaccccca atggtttaca aaagcaatgc
     1921 cctggtcgag gccgcgtatc gcctcagtgt tcaggaacag cggatcgttc tggcctgtat
     1981 tagccaggtg aagaggagcg agcctgtcac cgatgaagtg atgtattcag tgacggcgga
     2041 ggacatagcg acgatggcgg gtgtccctat cgaatcttcc tacaaccagc tcaaagaagc
     2101 ggccctgcgc ctgaaacggc gggaagtccg gttaacccaa gagcccaatg gcaaggggaa
     2161 aagaccgagt gtgatgatta ccggctgggt gcaaacaatc atctaccggg agggtgaggg
     2221 ccgtgtagaa ctcaggttca ccaaagacat gctgccgtac ctgacggaac tcaccaaaca
     2281 gttcaccaaa tacgccttgg ctgacgtggc caagatggac agcacccacg cgatcaggct
     2341 ttacgagctg ctcatgcaat gggacagcat cggccagcgc gaaatagaaa ttgaccagct
     2401 gcgaaagtgg tttcaactgg aaggccggta tccctcgatc aaggacttca agttgcgagt
     2461 gcttgatcca gccgtgacgc agatcaacga gcacagcccg ctacaggtgg agtgggcgca
     2521 gcgaaagacc gggcgcaagg tcacacatct gttgttcagt tttggaccga agaagcccgc
     2581 caaggcggtg ggtaaggccc cagcgaagcg caaggccggg aagatttcag atgctgagat
     2641 cgcgaaacag gctcgccctg gtgagacatg ggaagcggcc cgcgctcgac taacccagat
     2701 gccgctggat ctggcctaga ggccgtggcc accacggcca acagataaaa cgaaaggccc
     2761 agtctttcga ctgagccttt cgttttattt gatgcctggc agttccctac tctcgcatgg
     2821 ggagacccca cactaccatc ggcgctacgg cgtttcactt ctgagttcgg catggggtca
     2881 ggtgggacca ccgcgctact gccgccaggc aaattctgtt ttatcagacc gcttctgcgt
     2941 tctgatttaa tctgtatcag gctgaggtta attaagctgc gctagtagac gagtccatgt
     3001 gctggcgttc aaatttcgca gcagcggttt ctttaccaga ctcgagggta ccgacgtcag
     3061 cgatcgcgtg gccggccgat atccaattga gatctgccat atgtatagta tgtatatctc
     3121 cttcttaaag ttaaacaaaa ttatttctag cccaaaaaaa cgggtatgga gaaacagtag
     3181 agagttgcga taaaaagcgt caggtaggat ccgctaatct tatggataaa aatgctatgg
     3241 catagcaaag tgtgacgccg tgcaaataat caatgtggac ttttctgccg tgattataga
     3301 cacttttgtt acgcgttttt gtcatggctt tggtcccgct ttgttacaga atgcttttaa
     3361 taagcggggt taccggtttg gttagcgaga agagccagta aaagacgcag tgacggcaat
     3421 gtctgatgca atatggacaa ttggtttctt aacagataaa acgaaaggcc cagtctttcg
     3481 actgagcctt tcgttttatt tgatgcctgg cagttcccta ctctcgcatg gggagacccc
     3541 acactaccat cggcgctacg gcgtttcact tctgagttcg gcatggggtc aggtgggacc
     3601 accgcgctac tgccgccagg caaattctgt tttatttcga ttatgcggcc gtgtacaata
     3661 cgattacttt ctgttcgact taagcattat gcggccgcaa gcttgtcgac ctgcaggcgc
     3721 gccgagctcg aattcggatc ctggctgtgg tgatgatggt gatggctgct gcccatgggt
     3781 atgtatatct ccttcttaaa gttaaacaaa attatttcta gcccaaaaaa acgggtatgg
     3841 agaaacagta gagagttgcg ataaaaagcg tcaggtagga tccgctaatc ttatggataa
     3901 aaatgctatg gcatagcaaa gtgtgacgcc gtgcaaataa tcaatgtgga cttttctgcc
     3961 gtgattatag acacttttgt tacgcgtttt tgtcatggct ttggtcccgc tttgttacag
     4021 aatgctttta ataagcgggg ttaccggttt ggttagcgag aagagccagt aaaagacgca
     4081 gtgacggcaa tgtctgatgc aatatggaca attggtttct tctctgaatg gcgggagtat
     4141 gaaaagtatg gctgaagcgc aaaatgatcc cctgctgccg ggatactcgt ttaatgccca
     4201 tctggtggcg ggtttaacgc cgattgaggc caacggttat ctcgattttt ttatcgaccg
     4261 accgctggga atgaaaggtt atattctcaa tctcaccatt cgcggtcagg gggtggtgaa
     4321 aaatcaggga cgagaatttg tttgccgacc gggtgatatt ttgctgttcc cgccaggaga
     4381 gattcatcac tacggtcgtc atccggaggc tcgcgaatgg tatcaccagt gggtttactt
     4441 tcgtccgcgc gcctactggc atgaatggct taactggccg tcaatatttg ccaatacggg
     4501 gttctttcgc ccggatgaag cgcaccagcc gcatttcagc gacctgtttg ggcaaatcat
     4561 taacgccggg caaggggaag ggcgctattc ggagctgctg gcgataaatc tgcttgagca
     4621 attgttactg cggcgcatgg aagcgattaa cgagtcgctc catccaccga tggataatcg
     4681 ggtacgcgag gcttgtcagt acatcagcga tcacctggca gacagcaatt ttgatatcgc
     4741 cagcgtcgca cagcatgttt gcttgtcgcc gtcgcgtctg tcacatcttt tccgccagca
     4801 gttagggatt agcgtcttaa gctggcgcga ggaccaacgt atcagccagg cgaagctgct
     4861 tttgagcacc acccggatgc ctatcgccac cgtcggtcgc aatgttggtt ttgacgatca
     4921 actctatttc tcgcgggtat ttaaaaaatg caccggggcc agcccgagcg agttccgtgc
     4981 cggttgtgaa gaaaaagtga atgatgtagc cgtcaagttg tcataatttg gggattcctt
     5041 aaggtatact ttccgctgca taaccctgct tcggggtcat tatagcgatt ttttcggtat
     5101 atccatcctt tttcgcacga tatacaggat tttgccaaag ggttcgtgta gactttcctt
     5161 ggtgtatcca acggcgtcag ccgggcagga taggtgaagt aggcccaccc gcgagcgggt
     5221 gttccttctt cactgtccct tattcgcacc tggcggtgct caacgggaat cctgctctgc
     5281 gaggctggcc gataagctag cttatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     5341 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     5401 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     5461 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     5521 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     5581 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     5641 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     5701 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     5761 cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct
     5821 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     5881 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     5941 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     6001 actcacgtta agggattttg gt
//
