LOCUS       40924_24483        4993 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Escherichia-Pseudomonas shuttle vector pHERD28T, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4993)
  AUTHORS   Qiu D, Damron FH, Mima T, Schweizer HP, Yu HD.
  TITLE     PBAD-based shuttle vectors for functional analysis of toxic and 
            highly regulated genes in Pseudomonas and Burkholderia spp. and 
            other bacteria
  JOURNAL   Appl. Environ. Microbiol. 74 (23), 7422-7426 (2008)
  PUBMED    18849445
REFERENCE   2  (bases 1 to 4993)
  AUTHORS   Qiu D, Yu HD.
  TITLE     Direct Submission
  JOURNAL   Submitted (31-MAR-2008) Biochemistry and Microbiology, Marshall 
            University Joan C. Edwards School of Medicine, 1 John Marshall 
            Drive, Huntington, WV 25755, USA
REFERENCE   3  (bases 1 to 4993)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4993)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Appl. 
            Environ. Microbiol."; date: "2008"; volume: "74"; issue: "23"; 
            pages: "7422-7426"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (31-MAR-2008) Biochemistry and Microbiology, Marshall University 
            Joan C. Edwards School of Medicine, 1 John Marshall Drive, 
            Huntington, WV 25755, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4993
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(660..893)
                     /codon_start=1
                     /label=TpR
                     /note="E. coli plasmid-associated dihydrofolate reductase"
                     /translation="MGQSSDEANAPVAGQFALPLSATFGLGDRVRKKSGAAWQGQVVGW
                     YCTKLTPEGYAVESESHPGSVQIYPVAALERVA"
     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
                     /codon_start=1
                     /label=pRO1600 Rep
                     /note="replication protein for the broad-host-range plasmid
                     pRO1600 from Pseudomonas aeruginosa"
                     /translation="VASPPMVYKSNALVEAAYRLSVQEQRIVLACISQVKRSEPVTDEV
                     MYSVTAEDIATMAGVPIESSYNQLKEAALRLKRREVRLTQEPNGKGKRPSVMITGWVQT
                     IIYREGEGRVELRFTKDMLPYLTELTKQFTKYALADVAKMDSTHAIRLYELLMQWDSIG
                     QREIEIDQLRKWFQLEGRYPSIKDFKLRVLDPAVTQINEHSPLQVEWAQRKTGRKVTHL
                     LFSFGPKKPAKAVGKAPAKRKAGKISDAEIAKQARPGETWEAARARLTQMPLDLA"
     primer_bind     2657..2673
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    complement(2677..2733)
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     RBS             complement(2759..2781)
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     promoter        complement(2808..3092)
                     /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             3119..3994
                     /codon_start=1
                     /label=araC
                     /note="L-arabinose regulatory protein"
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     oriT            4154..4262
                     /label=oriT
                     /note="incP origin of transfer"
     rep_origin      complement(4334..4922)
                     /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 acggcccggc
     2521 ctgcctttca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta
     2581 cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt
     2641 tcccagtcac gacgttgtaa aacgacggcc agtgccaagc ttgcatgcct gcaggtcgac
     2701 tctagaggat ccccgggtac cgagctcgaa ttcttatcag atcccatggg tatgtatatc
     2761 tccttcttaa agttaaacaa aattatttct agcccaaaaa aacgggtatg gagaaacagt
     2821 agagagttgc gataaaaagc gtcaggtagg atccgctaat cttatggata aaaatgctat
     2881 ggcatagcaa agtgtgacgc cgtgcaaata atcaatgtgg acttttctgc cgtgattata
     2941 gacacttttg ttacgcgttt ttgtcatggc tttggtcccg ctttgttaca gaatgctttt
     3001 aataagcggg gttaccggtt tggttagcga gaagagccag taaaagacgc agtgacggca
     3061 atgtctgatg caatatggac aattggtttc ttctctgaat ggcgggagta tgaaaagtat
     3121 ggctgaagcg caaaatgatc ccctgctgcc gggatactcg tttaatgccc atctggtggc
     3181 gggtttaacg ccgattgagg ccaacggtta tctcgatttt tttatcgacc gaccgctggg
     3241 aatgaaaggt tatattctca atctcaccat tcgcggtcag ggggtggtga aaaatcaggg
     3301 acgagaattt gtttgccgac cgggtgatat tttgctgttc ccgccaggag agattcatca
     3361 ctacggtcgt catccggagg ctcgcgaatg gtatcaccag tgggtttact ttcgtccgcg
     3421 cgcctactgg catgaatggc ttaactggcc gtcaatattt gccaatacgg ggttctttcg
     3481 cccggatgaa gcgcaccagc cgcatttcag cgacctgttt gggcaaatca ttaacgccgg
     3541 gcaaggggaa gggcgctatt cggagctgct ggcgataaat ctgcttgagc aattgttact
     3601 gcggcgcatg gaagcgatta acgagtcgct ccatccaccg atggataatc gggtacgcga
     3661 ggcttgtcag tacatcagcg atcacctggc agacagcaat tttgatatcg ccagcgtcgc
     3721 acagcatgtt tgcttgtcgc cgtcgcgtct gtcacatctt ttccgccagc agttagggat
     3781 tagcgtctta agctggcgcg aggaccaacg tatcagccag gcgaagctgc ttttgagcac
     3841 cacccggatg cctatcgcca ccgtcggtcg caatgttggt tttgacgatc aactctattt
     3901 ctcgcgggta tttaaaaaat gcaccggggc cagcccgagc gagttccgtg ccggttgtga
     3961 agaaaaagtg aatgatgtag ccgtcaagtt gtcataattt ggggattcct taaggtatac
     4021 tttccgctgc ataaccctgc ttcggggtca ttatagcgat tttttcggta tatccatcct
     4081 ttttcgcacg atatacagga ttttgccaaa gggttcgtgt agactttcct tggtgtatcc
     4141 aacggcgtca gccgggcagg ataggtgaag taggcccacc cgcgagcggg tgttccttct
     4201 tcactgtccc ttattcgcac ctggcggtgc tcaacgggaa tcctgctctg cgaggctggc
     4261 cgataagcta gcttatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     4321 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     4381 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     4441 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     4501 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     4561 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     4621 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     4681 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     4741 tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc
     4801 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     4861 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     4921 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     4981 aagggatttt ggt
//
