LOCUS       40924_16300        6972 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Bicistronic shuttle vector pDUO-26T, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6972)
  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 6972)
  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 6972)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6972)
  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..6972
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(12..30)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(48..64)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(72..88)
                     /label=lac operator
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(96..126)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(141..162)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        409..437
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein
                     gene"
     CDS             485..1672
                     /label=TcR
                     /note="tetracycline efflux protein"
     rep_origin      2497..3085
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     oriT            complement(3157..3265)
                     /direction=LEFT
                     /label=oriT
                     /note="incP origin of transfer"
     CDS             complement(3425..4300)
                     /label=araC
                     /note="L-arabinose regulatory protein"
     promoter        4327..4611
                     /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             4638..4660
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             4684..4701
                     /label=6xHis
                     /note="6xHis affinity tag"
     terminator      4946..4992
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     promoter        4998..5282
                     /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    5001..5016
                     /label=O2 region
                     /note="O2 region"
     misc_feature    5158..5179
                     /label=O1 region
                     /note="O1 region"
     misc_feature    5200..5213
                     /label=CAP binding site
                     /note="CAP binding site"
     misc_feature    5210..5248
                     /label=I2 and I1 region
                     /note="I2 and I1 region"
     RBS             5309..5331
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     misc_feature    5325..5330
                     /label=RBS
                     /note="RBS"
     CDS             5414..5458
                     /label=S-Tag
                     /note="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
     terminator      complement(5660..5703)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     CDS             complement(5732..6562)
                     /label=pRO1600 Rep
                     /note="replication protein for the broad-host-range plasmid
                     pRO1600 from Pseudomonas aeruginosa"
     rep_origin      6576..6927
                     /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 agcttgagta ttctatagtg tcacctaaat agcttggcgt aatcatggtc atagctgttt
       61 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
      121 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
      181 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
      241 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
      301 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
      361 acagaatcag gggataacgc aggaaagaac atgaattaat tctcatgttt gacagcttat
      421 catcgattag ctttaatgcg gtagtttatc acagttaaat tgctaacgca gtcaggcacc
      481 gtgtatgaaa tctaacaatg cgctcatcgt catcctcggc accgtcaccc tggatgctgt
      541 aggcataggc ttggttatgc cggtactgcc gggcctcttg cgggatatcg tccattccga
      601 cagcatcgcc agtcactatg gcgtgctgct agcgctatat gcgttgatgc aatttctatg
      661 cgcacccgtt ctcggagcac tgtccgaccg ctttggccgc cgcccagtcc tgctcgcttc
      721 gctacttgga gccactatcg actacgcgat catggcgacc acacccgtcc tgtggattct
      781 ctacgccgga cgcatcgtgg ccggcatcac cggcgccaca ggtgcggttg ctggcgccta
      841 tatcgccgac atcaccgatg gggaagatcg ggctcgccac ttcgggctca tgagcgcttg
      901 tttcggcgtg ggtatggtgg caggccccgt ggccggggga ctgttgggcg ccatctcctt
      961 acatgcacca ttccttgcgg cggcggtgct caacggcctc aacctactac tgggctgctt
     1021 cctaatgcag gagtcgcata agggagagcg ccgaccgatg cccttgagag ccttcaaccc
     1081 agtcagctcc ttccggtggg cgcggggcat gactatcgtc gccgcactta tgactgtctt
     1141 ctttatcatg caactcgtag gacaggtgcc ggcagcgctc tgggtcattt tcggcgagga
     1201 ccgctttcgc tggagcgcga cgatgatcgg cctgtcgctt gcggtattcg gaatcttgca
     1261 cgccctcgct caagccttcg tcactggtcc cgccaccaaa cgtttcggcg agaagcaggc
     1321 cattatcgcc ggcatggcgg ccgacgcgct gggctacgtc ttgctggcgt tcgcgacgcg
     1381 aggctggatg gccttcccca ttatgattct tctcgcttcc ggcggcatcg ggatgcccgc
     1441 gttgcaggcc atgctgtcca ggcaggtaga tgacgaccat cagggacagc ttcaaggatc
     1501 gctcgcggct cttaccagcc taacttcgat cactggaccg ctgatcgtca cggcgattta
     1561 tgccgcctcg gcgagcacat ggaacgggtt ggcatggatt gtaggcgccg ccctatacct
     1621 tgtctgcctc cccgcgttgc gtcgcggtgc atggagccgg gccacctcga cctgaatgga
     1681 agccggcggc acctcgctaa cggattcacc actccaagaa ttggagccaa tcaattcttg
     1741 cggagaactg tgaatgcgca aaccaaccct tgactcacca gtcacagaaa agcatcttac
     1801 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     1861 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     1921 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     1981 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     2041 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     2101 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     2161 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     2221 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     2281 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     2341 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     2401 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     2461 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     2521 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     2581 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     2641 tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     2701 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     2761 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     2821 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     2881 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     2941 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     3001 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     3061 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     3121 cttttgctgg ccttttgctc acataagcta gcttatcggc cagcctcgca gagcaggatt
     3181 cccgttgagc accgccaggt gcgaataagg gacagtgaag aaggaacacc cgctcgcggg
     3241 tgggcctact tcacctatcc tgcccggctg acgccgttgg atacaccaag gaaagtctac
     3301 acgaaccctt tggcaaaatc ctgtatatcg tgcgaaaaag gatggatata ccgaaaaaat
     3361 cgctataatg accccgaagc agggttatgc agcggaaagt ataccttaag gaatccccaa
     3421 attatgacaa cttgacggct acatcattca ctttttcttc acaaccggca cggaactcgc
     3481 tcgggctggc cccggtgcat tttttaaata cccgcgagaa atagagttga tcgtcaaaac
     3541 caacattgcg accgacggtg gcgataggca tccgggtggt gctcaaaagc agcttcgcct
     3601 ggctgatacg ttggtcctcg cgccagctta agacgctaat ccctaactgc tggcggaaaa
     3661 gatgtgacag acgcgacggc gacaagcaaa catgctgtgc gacgctggcg atatcaaaat
     3721 tgctgtctgc caggtgatcg ctgatgtact gacaagcctc gcgtacccga ttatccatcg
     3781 gtggatggag cgactcgtta atcgcttcca tgcgccgcag taacaattgc tcaagcagat
     3841 ttatcgccag cagctccgaa tagcgccctt ccccttgccc ggcgttaatg atttgcccaa
     3901 acaggtcgct gaaatgcggc tggtgcgctt catccgggcg aaagaacccc gtattggcaa
     3961 atattgacgg ccagttaagc cattcatgcc agtaggcgcg cggacgaaag taaacccact
     4021 ggtgatacca ttcgcgagcc tccggatgac gaccgtagtg atgaatctct cctggcggga
     4081 acagcaaaat atcacccggt cggcaaacaa attctcgtcc ctgatttttc accaccccct
     4141 gaccgcgaat ggtgagattg agaatataac ctttcattcc cagcggtcgg tcgataaaaa
     4201 aatcgagata accgttggcc tcaatcggcg ttaaacccgc caccagatgg gcattaaacg
     4261 agtatcccgg cagcagggga tcattttgcg cttcagccat acttttcata ctcccgccat
     4321 tcagagaaga aaccaattgt ccatattgca tcagacattg ccgtcactgc gtcttttact
     4381 ggctcttctc gctaaccaaa ccggtaaccc cgcttattaa aagcattctg taacaaagcg
     4441 ggaccaaagc catgacaaaa acgcgtaaca aaagtgtcta taatcacggc agaaaagtcc
     4501 acattgatta tttgcacggc gtcacacttt gctatgccat agcattttta tccataagat
     4561 tagcggatcc tacctgacgc tttttatcgc aactctctac tgtttctcca tacccgtttt
     4621 tttgggctag aaataatttt gtttaacttt aagaaggaga tatacatacc catgggcagc
     4681 agccatcacc atcatcacca cagccaggat ccgaattcga gctcggcgcg cctgcaggtc
     4741 gacaagcttg cggccgcata atgcttaagt cgaacagaaa gtaatcgtat tgtacacggc
     4801 cgcataatcg aaataaaaca gaatttgcct ggcggcagta gcgcggtggt cccacctgac
     4861 cccatgccga actcagaagt gaaacgccgt agcgccgatg gtagtgtggg gtctccccat
     4921 gcgagagtag ggaactgcca ggcatcaaat aaaacgaaag gctcagtcga aagactgggc
     4981 ctttcgtttt atctgttaag aaaccaattg tccatattgc atcagacatt gccgtcactg
     5041 cgtcttttac tggctcttct cgctaaccaa accggtaacc ccgcttatta aaagcattct
     5101 gtaacaaagc gggaccaaag ccatgacaaa aacgcgtaac aaaagtgtct ataatcacgg
     5161 cagaaaagtc cacattgatt atttgcacgg cgtcacactt tgctatgcca tagcattttt
     5221 atccataaga ttagcggatc ctacctgacg ctttttatcg caactctcta ctgtttctcc
     5281 atacccgttt ttttgggcta gaaataattt tgtttaactt taagaaggag atatacatac
     5341 tatacatatg gcagatctca attggatatc ggccggccac gcgatcgctg acgtcggtac
     5401 cctcgagtct ggtaaagaaa ccgctgctgc gaaatttgaa cgccagcaca tggactcgtc
     5461 tactagcgca gcttaattaa cctcagcctg atacagatta aatcagaacg cagaagcggt
     5521 ctgataaaac agaatttgcc tggcggcagt agcgcggtgg tcccacctga ccccatgccg
     5581 aactcagaag tgaaacgccg tagcgccgat ggtagtgtgg ggtctcccca tgcgagagta
     5641 gggaactgcc aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt
     5701 tatctgttgg ccgtggtggc cacggcctct aggccagatc cagcggcatc tgggttagtc
     5761 gagcgcgggc cgcttcccat gtctcaccag ggcgagcctg tttcgcgatc tcagcatctg
     5821 aaatcttccc ggccttgcgc ttcgctgggg ccttacccac cgccttggcg ggcttcttcg
     5881 gtccaaaact gaacaacaga tgtgtgacct tgcgcccggt ctttcgctgc gcccactcca
     5941 cctgtagcgg gctgtgctcg ttgatctgcg tcacggctgg atcaagcact cgcaacttga
     6001 agtccttgat cgagggatac cggccttcca gttgaaacca ctttcgcagc tggtcaattt
     6061 ctatttcgcg ctggccgatg ctgtcccatt gcatgagcag ctcgtaaagc ctgatcgcgt
     6121 gggtgctgtc catcttggcc acgtcagcca aggcgtattt ggtgaactgt ttggtgagtt
     6181 ccgtcaggta cggcagcatg tctttggtga acctgagttc tacacggccc tcaccctccc
     6241 ggtagatgat tgtttgcacc cagccggtaa tcatcacact cggtcttttc cccttgccat
     6301 tgggctcttg ggttaaccgg acttcccgcc gtttcaggcg cagggccgct tctttgagct
     6361 ggttgtagga agattcgata gggacacccg ccatcgtcgc tatgtcctcc gccgtcactg
     6421 aatacatcac ttcatcggtg acaggctcgc tcctcttcac ctggctaata caggccagaa
     6481 cgatccgctg ttcctgaaca ctgaggcgat acgcggcctc gaccagggca ttgcttttgt
     6541 aaaccattgg gggtgaggcc acgttcgaca ttccttgtgt ataaggggac actgtatctg
     6601 cgtcccacaa tacaacaaat ccgtcccttt acaacaacaa atccgtccct tcttaacaac
     6661 aaatccgtcc cttaatggca acaaatccgt ccctttttaa actctacagg ccacggatta
     6721 cgtggcctgt agacgtccta aaaggtttaa aagggaaaag gaagaaaagg gtggaaacgc
     6781 aaaaaacgca ccactacgtg gccccgttgg ggccgcattt gtgcccctga aggggcgggg
     6841 gaggcgtctg ggcaatcccc gttttaccag tcccctatcg ccgcctgaga gggcgcagga
     6901 agcgagtaat cagggtatcg aggcggattc acccttggcg tccaaccagc ggcaccagcg
     6961 gcgcctgaga gg
//
