LOCUS       40924_16295        5893 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Bicistronic shuttle vector pDUO-20T, complete sequence.
ACCESSION   .
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 Children's Nutrition 
            Research Center, Baylor College of Medicine, 1100 Bates Street, 
            Houston, TX 77030-2600, USA
REFERENCE   3  (bases 1 to 5893)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5893)
  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..5893
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        96..200
                     /label=AmpR promoter
     CDS             201..1058
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      1232..1820
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     oriT            complement(1892..2000)
                     /direction=LEFT
                     /label=oriT
                     /note="incP origin of transfer"
     CDS             complement(2160..3035)
                     /label=araC
                     /note="L-arabinose regulatory protein"
     promoter        3062..3346
                     /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)"
     RBS             3373..3395
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             3419..3436
                     /label=6xHis
                     /note="6xHis affinity tag"
     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"
     CDS             4149..4193
                     /label=S-Tag
                     /note="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
     terminator      complement(4395..4438)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     CDS             complement(4467..5297)
                     /label=pRO1600 Rep
                     /note="replication protein for the broad-host-range plasmid
                     pRO1600 from Pseudomonas aeruginosa"
     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)"
ORIGIN
        1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
       61 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
      121 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
      181 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
      241 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
      301 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
      361 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
      421 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
      481 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
      541 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
      601 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
      661 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
      721 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
      781 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
      841 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
      901 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
      961 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     1021 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     1081 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     1141 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     1201 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     1261 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     1321 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc
     1381 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     1441 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     1501 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     1561 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     1621 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     1681 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     1741 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     1801 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     1861 gctggccttt tgctcacata agctagctta tcggccagcc tcgcagagca ggattcccgt
     1921 tgagcaccgc caggtgcgaa taagggacag tgaagaagga acacccgctc gcgggtgggc
     1981 ctacttcacc tatcctgccc ggctgacgcc gttggataca ccaaggaaag tctacacgaa
     2041 ccctttggca aaatcctgta tatcgtgcga aaaaggatgg atataccgaa aaaatcgcta
     2101 taatgacccc gaagcagggt tatgcagcgg aaagtatacc ttaaggaatc cccaaattat
     2161 gacaacttga cggctacatc attcactttt tcttcacaac cggcacggaa ctcgctcggg
     2221 ctggccccgg tgcatttttt aaatacccgc gagaaataga gttgatcgtc aaaaccaaca
     2281 ttgcgaccga cggtggcgat aggcatccgg gtggtgctca aaagcagctt cgcctggctg
     2341 atacgttggt cctcgcgcca gcttaagacg ctaatcccta actgctggcg gaaaagatgt
     2401 gacagacgcg acggcgacaa gcaaacatgc tgtgcgacgc tggcgatatc aaaattgctg
     2461 tctgccaggt gatcgctgat gtactgacaa gcctcgcgta cccgattatc catcggtgga
     2521 tggagcgact cgttaatcgc ttccatgcgc cgcagtaaca attgctcaag cagatttatc
     2581 gccagcagct ccgaatagcg cccttcccct tgcccggcgt taatgatttg cccaaacagg
     2641 tcgctgaaat gcggctggtg cgcttcatcc gggcgaaaga accccgtatt ggcaaatatt
     2701 gacggccagt taagccattc atgccagtag gcgcgcggac gaaagtaaac ccactggtga
     2761 taccattcgc gagcctccgg atgacgaccg tagtgatgaa tctctcctgg cgggaacagc
     2821 aaaatatcac ccggtcggca aacaaattct cgtccctgat ttttcaccac cccctgaccg
     2881 cgaatggtga gattgagaat ataacctttc attcccagcg gtcggtcgat aaaaaaatcg
     2941 agataaccgt tggcctcaat cggcgttaaa cccgccacca gatgggcatt aaacgagtat
     3001 cccggcagca ggggatcatt ttgcgcttca gccatacttt tcatactccc gccattcaga
     3061 gaagaaacca attgtccata ttgcatcaga cattgccgtc actgcgtctt ttactggctc
     3121 ttctcgctaa ccaaaccggt aaccccgctt attaaaagca ttctgtaaca aagcgggacc
     3181 aaagccatga caaaaacgcg taacaaaagt gtctataatc acggcagaaa agtccacatt
     3241 gattatttgc acggcgtcac actttgctat gccatagcat ttttatccat aagattagcg
     3301 gatcctacct gacgcttttt atcgcaactc tctactgttt ctccataccc gtttttttgg
     3361 gctagaaata attttgttta actttaagaa ggagatatac atacccatgg gcagcagcca
     3421 tcaccatcat caccacagcc aggatccgaa ttcgagctcg gcgcgcctgc aggtcgacaa
     3481 gcttgcggcc gcataatgct taagtcgaac agaaagtaat cgtattgtac acggccgcat
     3541 aatcgaaata aaacagaatt tgcctggcgg cagtagcgcg gtggtcccac ctgaccccat
     3601 gccgaactca gaagtgaaac gccgtagcgc cgatggtagt gtggggtctc cccatgcgag
     3661 agtagggaac tgccaggcat caaataaaac gaaaggctca gtcgaaagac tgggcctttc
     3721 gttttatctg ttaagaaacc aattgtccat attgcatcag acattgccgt cactgcgtct
     3781 tttactggct cttctcgcta accaaaccgg taaccccgct tattaaaagc attctgtaac
     3841 aaagcgggac caaagccatg acaaaaacgc gtaacaaaag tgtctataat cacggcagaa
     3901 aagtccacat tgattatttg cacggcgtca cactttgcta tgccatagca tttttatcca
     3961 taagattagc ggatcctacc tgacgctttt tatcgcaact ctctactgtt tctccatacc
     4021 cgtttttttg ggctagaaat aattttgttt aactttaaga aggagatata catactatac
     4081 atatggcaga tctcaattgg atatcggccg gccacgcgat cgctgacgtc ggtaccctcg
     4141 agtctggtaa agaaaccgct gctgcgaaat ttgaacgcca gcacatggac tcgtctacta
     4201 gcgcagctta attaacctca gcctgataca gattaaatca gaacgcagaa gcggtctgat
     4261 aaaacagaat ttgcctggcg gcagtagcgc ggtggtccca cctgacccca tgccgaactc
     4321 agaagtgaaa cgccgtagcg ccgatggtag tgtggggtct ccccatgcga gagtagggaa
     4381 ctgccaggca tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt cgttttatct
     4441 gttggccgtg gtggccacgg cctctaggcc agatccagcg gcatctgggt tagtcgagcg
     4501 cgggccgctt cccatgtctc accagggcga gcctgtttcg cgatctcagc atctgaaatc
     4561 ttcccggcct tgcgcttcgc tggggcctta cccaccgcct tggcgggctt cttcggtcca
     4621 aaactgaaca acagatgtgt gaccttgcgc ccggtctttc gctgcgccca ctccacctgt
     4681 agcgggctgt gctcgttgat ctgcgtcacg gctggatcaa gcactcgcaa cttgaagtcc
     4741 ttgatcgagg gataccggcc ttccagttga aaccactttc gcagctggtc aatttctatt
     4801 tcgcgctggc cgatgctgtc ccattgcatg agcagctcgt aaagcctgat cgcgtgggtg
     4861 ctgtccatct tggccacgtc agccaaggcg tatttggtga actgtttggt gagttccgtc
     4921 aggtacggca gcatgtcttt ggtgaacctg agttctacac ggccctcacc ctcccggtag
     4981 atgattgttt gcacccagcc ggtaatcatc acactcggtc ttttcccctt gccattgggc
     5041 tcttgggtta accggacttc ccgccgtttc aggcgcaggg ccgcttcttt gagctggttg
     5101 taggaagatt cgatagggac acccgccatc gtcgctatgt cctccgccgt cactgaatac
     5161 atcacttcat cggtgacagg ctcgctcctc ttcacctggc taatacaggc cagaacgatc
     5221 cgctgttcct gaacactgag gcgatacgcg gcctcgacca gggcattgct tttgtaaacc
     5281 attgggggtg aggccacgtt cgacattcct tgtgtataag gggacactgt atctgcgtcc
     5341 cacaatacaa caaatccgtc cctttacaac aacaaatccg tcccttctta acaacaaatc
     5401 cgtcccttaa tggcaacaaa tccgtccctt tttaaactct acaggccacg gattacgtgg
     5461 cctgtagacg tcctaaaagg tttaaaaggg aaaaggaaga aaagggtgga aacgcaaaaa
     5521 acgcaccact acgtggcccc gttggggccg catttgtgcc cctgaagggg cgggggaggc
     5581 gtctgggcaa tccccgtttt accagtcccc tatcgccgcc tgagagggcg caggaagcga
     5641 gtaatcaggg tatcgaggcg gattcaccct tggcgtccaa ccagcggcac cagcggcgcc
     5701 tgagaggtat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc
     5761 cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct
     5821 tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca
     5881 ccgaaacgcg cga
//
