LOCUS       40924_24473        5087 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Escherichia-Pseudomonas shuttle vector pHERD20T, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5087)
  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 5087)
  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 5087)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5087)
  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..5087
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        96..200
                     /label=AmpR promoter
     CDS             201..1058
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     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)
                     /codon_start=1
                     /label=araC
                     /note="L-arabinose regulatory protein"
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     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)"
     misc_feature    3421..3477
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(3481..3497)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(3661..4491)
                     /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"
     rep_origin      4505..4856
                     /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 gatctgataa
     3421 gaattcgagc tcggtacccg gggatcctct agagtcgacc tgcaggcatg caagcttggc
     3481 actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg
     3541 ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg
     3601 cccttcccaa cagttgcgca gcctgaaagg caggccgggc cgtggtggcc acggcctcta
     3661 ggccagatcc agcggcatct gggttagtcg agcgcgggcc gcttcccatg tctcaccagg
     3721 gcgagcctgt ttcgcgatct cagcatctga aatcttcccg gccttgcgct tcgctggggc
     3781 cttacccacc gccttggcgg gcttcttcgg tccaaaactg aacaacagat gtgtgacctt
     3841 gcgcccggtc tttcgctgcg cccactccac ctgtagcggg ctgtgctcgt tgatctgcgt
     3901 cacggctgga tcaagcactc gcaacttgaa gtccttgatc gagggatacc ggccttccag
     3961 ttgaaaccac tttcgcagct ggtcaatttc tatttcgcgc tggccgatgc tgtcccattg
     4021 catgagcagc tcgtaaagcc tgatcgcgtg ggtgctgtcc atcttggcca cgtcagccaa
     4081 ggcgtatttg gtgaactgtt tggtgagttc cgtcaggtac ggcagcatgt ctttggtgaa
     4141 cctgagttct acacggccct caccctcccg gtagatgatt gtttgcaccc agccggtaat
     4201 catcacactc ggtcttttcc ccttgccatt gggctcttgg gttaaccgga cttcccgccg
     4261 tttcaggcgc agggccgctt ctttgagctg gttgtaggaa gattcgatag ggacacccgc
     4321 catcgtcgct atgtcctccg ccgtcactga atacatcact tcatcggtga caggctcgct
     4381 cctcttcacc tggctaatac aggccagaac gatccgctgt tcctgaacac tgaggcgata
     4441 cgcggcctcg accagggcat tgcttttgta aaccattggg ggtgaggcca cgttcgacat
     4501 tccttgtgta taaggggaca ctgtatctgc gtcccacaat acaacaaatc cgtcccttta
     4561 caacaacaaa tccgtccctt cttaacaaca aatccgtccc ttaatggcaa caaatccgtc
     4621 cctttttaaa ctctacaggc cacggattac gtggcctgta gacgtcctaa aaggtttaaa
     4681 agggaaaagg aagaaaaggg tggaaacgca aaaaacgcac cactacgtgg ccccgttggg
     4741 gccgcatttg tgcccctgaa ggggcggggg aggcgtctgg gcaatccccg ttttaccagt
     4801 cccctatcgc cgcctgagag ggcgcaggaa gcgagtaatc agggtatcga ggcggattca
     4861 cccttggcgt ccaaccagcg gcaccagcgg cgcctgagag gtatggtgca ctctcagtac
     4921 aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc
     4981 gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg
     5041 gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcga
//
