LOCUS       Exported                5084 bp DNA     circular SYN 19-JUL-2025
DEFINITION  Escherichia-Pseudomonas shuttle vector pHERD20T, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5084)
  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 5084)
  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 5084)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5084)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 5084)
  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"
COMMENT     SGRef: number: 4; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5084
                     /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(1890..1998)
                     /direction=LEFT
                     /label=oriT
                     /note="incP origin of transfer"
     CDS             complement(2158..3033)
                     /codon_start=1
                     /label=araC
                     /note="L-arabinose regulatory protein"
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        3060..3344
                     /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             3371..3393
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     misc_feature    3419..3475
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(3479..3495)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(3659..4489)
                     /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      4503..4854
                     /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 aatactgttc
     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 tgctcacaag ctagcttatc ggccagcctc gcagagcagg attcccgttg
     1921 agcaccgcca ggtgcgaata agggacagtg aagaaggaac acccgctcgc gggtgggcct
     1981 acttcaccta tcctgcccgg ctgacgccgt tggatacacc aaggaaagtc tacacgaacc
     2041 ctttggcaaa atcctgtata tcgtgcgaaa aaggatggat ataccgaaaa aatcgctata
     2101 atgaccccga agcagggtta tgcagcggaa agtatacctt aaggaatccc caaattatga
     2161 caacttgacg gctacatcat tcactttttc ttcacaaccg gcacggaact cgctcgggct
     2221 ggccccggtg cattttttaa atacccgcga gaaatagagt tgatcgtcaa aaccaacatt
     2281 gcgaccgacg gtggcgatag gcatccgggt ggtgctcaaa agcagcttcg cctggctgat
     2341 acgttggtcc tcgcgccagc ttaagacgct aatccctaac tgctggcgga aaagatgtga
     2401 cagacgcgac ggcgacaagc aaacatgctg tgcgacgctg gcgatatcaa aattgctgtc
     2461 tgccaggtga tcgctgatgt actgacaagc ctcgcgtacc cgattatcca tcggtggatg
     2521 gagcgactcg ttaatcgctt ccatgcgccg cagtaacaat tgctcaagca gatttatcgc
     2581 cagcagctcc gaatagcgcc cttccccttg cccggcgtta atgatttgcc caaacaggtc
     2641 gctgaaatgc ggctggtgcg cttcatccgg gcgaaagaac cccgtattgg caaatattga
     2701 cggccagtta agccattcat gccagtaggc gcgcggacga aagtaaaccc actggtgata
     2761 ccattcgcga gcctccggat gacgaccgta gtgatgaatc tctcctggcg ggaacagcaa
     2821 aatatcaccc ggtcggcaaa caaattctcg tccctgattt ttcaccaccc cctgaccgcg
     2881 aatggtgaga ttgagaatat aacctttcat tcccagcggt cggtcgataa aaaaatcgag
     2941 ataaccgttg gcctcaatcg gcgttaaacc cgccaccaga tgggcattaa acgagtatcc
     3001 cggcagcagg ggatcatttt gcgcttcagc catacttttc atactcccgc cattcagaga
     3061 agaaaccaat tgtccatatt gcatcagaca ttgccgtcac tgcgtctttt actggctctt
     3121 ctcgctaacc aaaccggtaa ccccgcttat taaaagcatt ctgtaacaaa gcgggaccaa
     3181 agccatgaca aaaacgcgta acaaaagtgt ctataatcac ggcagaaaag tccacattga
     3241 ttatttgcac ggcgtcacac tttgctatgc catagcattt ttatccataa gattagcgga
     3301 tcctacctga cgctttttat cgcaactctc tactgtttct ccatacccgt ttttttgggc
     3361 tagaaataat tttgtttaac tttaagaagg agatatacat acccatggga tctgataaga
     3421 attcgagctc ggtacccggg gatcctctag agtcgacctg caggcatgca agcttggcac
     3481 tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc
     3541 ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc
     3601 cttcccaaca gttgcgcagc ctgaaaggca ggccgggccg tggtggccac ggcctctagg
     3661 ccagatccag cggcatctgg gttagtcgag cgcgggccgc ttcccatgtc tcaccagggc
     3721 gagcctgttt cgcgatctca gcatctgaaa tcttcccggc cttgcgcttc gctggggcct
     3781 tacccaccgc cttggcgggc ttcttcggtc caaaactgaa caacagatgt gtgaccttgc
     3841 gcccggtctt tcgctgcgcc cactccacct gtagcgggct gtgctcgttg atctgcgtca
     3901 cggctggatc aagcactcgc aacttgaagt ccttgatcga gggataccgg ccttccagtt
     3961 gaaaccactt tcgcagctgg tcaatttcta tttcgcgctg gccgatgctg tcccattgca
     4021 tgagcagctc gtaaagcctg atcgcgtggg tgctgtccat cttggccacg tcagccaagg
     4081 cgtatttggt gaactgtttg gtgagttccg tcaggtacgg cagcatgtct ttggtgaacc
     4141 tgagttctac acggccctca ccctcccggt agatgattgt ttgcacccag ccggtaatca
     4201 tcacactcgg tcttttcccc ttgccattgg gctcttgggt taaccggact tcccgccgtt
     4261 tcaggcgcag ggccgcttct ttgagctggt tgtaggaaga ttcgataggg acacccgcca
     4321 tcgtcgctat gtcctccgcc gtcactgaat acatcacttc atcggtgaca ggctcgctcc
     4381 tcttcacctg gctaatacag gccagaacga tccgctgttc ctgaacactg aggcgatacg
     4441 cggcctcgac cagggcattg cttttgtaaa ccattggggg tgaggccacg ttcgacattc
     4501 cttgtgtata aggggacact gtatctgcgt cccacaatac aacaaatccg tccctttaca
     4561 acaacaaatc cgtcccttct taacaacaaa tccgtccctt aatggcaaca aatccgtccc
     4621 tttttaaact ctacaggcca cggattacgt ggcctgtaga cgtcctaaaa ggtttaaaag
     4681 ggaaaaggaa gaaaagggtg gaaacgcaaa aaacgcacca ctacgtggcc ccgttggggc
     4741 cgcatttgtg cccctgaagg ggcgggggag gcgtctgggc aatccccgtt ttaccagtcc
     4801 cctatcgccg cctgagaggg cgcaggaagc gagtaatcag ggtatcgagg cggattcacc
     4861 cttggcgtcc aaccagcggc accagcggcg cctgagagga tggtgcactc tcagtacaat
     4921 ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc
     4981 ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag
     5041 ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcga
//
