LOCUS       Exported                5214 bp DNA     circular SYN 19-JUL-2025
DEFINITION  Escherichia-Pseudomonas shuttle vector pHERD30T, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5214)
  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 5214)
  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 5214)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5214)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 5214)
  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..5214
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(733..1263)
                     /codon_start=1
                     /label=GmR
                     /note="gentamycin acetyltransferase"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(1452..1480)
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     rep_origin      complement(1521..1872)
                     /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             1886..2716
                     /codon_start=1
                     /label=pRO1600 Rep
                     /note="replication protein for the broad-host-range plasmid
                     pRO1600 from Pseudomonas aeruginosa"
                     /translation="VASPPMVYKSNALVEAAYRLSVQEQRIVLACISQVKRSEPVTDEV
                     MYSVTAEDIATMAGVPIESSYNQLKEAALRLKRREVRLTQEPNGKGKRPSVMITGWVQT
                     IIYREGEGRVELRFTKDMLPYLTELTKQFTKYALADVAKMDSTHAIRLYELLMQWDSIG
                     QREIEIDQLRKWFQLEGRYPSIKDFKLRVLDPAVTQINEHSPLQVEWAQRKTGRKVTHL
                     LFSFGPKKPAKAVGKAPARRKAGKISDAEIAKQARPGETWEAARARLTQMPLDLA"
     primer_bind     2880..2896
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    complement(2900..2956)
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     RBS             complement(2982..3004)
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     promoter        complement(3031..3315)
                     /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             3342..4217
                     /codon_start=1
                     /label=araC
                     /note="L-arabinose regulatory protein"
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     oriT            4377..4485
                     /label=oriT
                     /note="incP origin of transfer"
     rep_origin      complement(4555..5143)
                     /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 tgacctacgc 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 taatcctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtcgaattg
      661 gccgcggcgt tgtgacaatt taccgaacaa ctccgcggcc gggaagccga tctcggcttg
      721 aacgaattgt taggtggcgg tacttgggtc gatatcaaag tgcatcactt cttcccgtat
      781 gcccaacttt gtatagagag ccactgcggg atcgtcaccg taatctgctt gcacgtagat
      841 cacataagca ccaagcgcgt tggcctcatg cttgaggaga ttgatgagcg cggtggcaat
      901 gccctgcctc cggtgctcgc cggagactgc gagatcatag atatagatct cactacgcgg
      961 ctgctcaaac ttgggcagaa cgtaagccgc gagagcgcca acaaccgctt cttggtcgaa
     1021 ggcagcaagc gcgatgaatg tcttactacg gagcaagttc ccgaggtaat cggagtccgg
     1081 ctgatgttgg gagtaggtgg ctacgtctcc gaactcacga ccgaaaagat caagagcagc
     1141 ccgcatggat ttgacttggt cagggccgag cctacatgtg cgaatgatgc ccatacttga
     1201 gccacctaac tttgttttag ggcgactgcc ctgctgcgta acatcgttgc tgctgcgtaa
     1261 catcgttgct gctccataac atcaaacatc gacccacggc gtaacgcgct tgctgcttgg
     1321 atgcccgagg catagactgt acaaaaaaac agtcataaca agccatgaaa accgccactg
     1381 cgccgttacc accgctgcgt tcggtcaagg ttctggacca gttgcgtgag cgcatacgct
     1441 acttgcatta cagtttacga accgaacagg cttatgtcaa ttcgcgccgc tggtgccgct
     1501 ggttggacgc caagggtgaa tccgcctcga taccctgatt actcgcttcc tgcgccctct
     1561 caggcggcga taggggactg gtaaaacggg gattgcccag acgcctcccc cgccccttca
     1621 ggggcacaaa tgcggcccca acggggccac gtagtggtgc gttttttgcg tttccaccct
     1681 tttcttcctt ttccctttta aaccttttag gacgtctaca ggccacgtaa tccgtggcct
     1741 gtagagttta aaaagggacg gatttgttgc cattaaggga cggatttgtt gttaagaagg
     1801 gacggatttg ttgttgtaaa gggacggatt tgttgtattg tgggacgcag atacagtgtc
     1861 cccttataca caaggaatgt cgaacgtggc ctcaccccca atggtttaca aaagcaatgc
     1921 cctggtcgag gccgcgtatc gcctcagtgt tcaggaacag cggatcgttc tggcctgtat
     1981 tagccaggtg aagaggagcg agcctgtcac cgatgaagtg atgtattcag tgacggcgga
     2041 ggacatagcg acgatggcgg gtgtccctat cgaatcttcc tacaaccagc tcaaagaagc
     2101 ggccctgcgc ctgaaacggc gggaagtccg gttaacccaa gagcccaatg gcaaggggaa
     2161 aagaccgagt gtgatgatta ccggctgggt gcaaacaatc atctaccggg agggtgaggg
     2221 ccgtgtagaa ctcaggttca ccaaagacat gctgccgtac ctgacggaac tcaccaaaca
     2281 gttcaccaaa tacgccttgg ctgacgtggc caagatggac agcacccacg cgatcaggct
     2341 ttacgagctg ctcatgcaat gggacagcat cggccagcgc gaaatagaaa ttgaccagct
     2401 gcgaaagtgg tttcaactgg aaggccggta tccctcgatc aaggacttca agttgcgagt
     2461 gcttgatcca gccgtgacgc agatcaacga gcacagcccg ctacaggtgg agtgggcgca
     2521 gcgaaagacc gggcgcaagg tcacacatct gttgttcagt tttggaccga agaagcccgc
     2581 caaggcggtg ggtaaggccc cagcgaggcg caaggccggg aagatttcag atgctgagat
     2641 cgcgaaacag gctcgccctg gtgagacatg ggaagcggcc cgcgctcgac taacccagat
     2701 gccgctggat ctggcctaga ggccgtggcc accacggccc ggcctgcctt tcaggctgcg
     2761 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
     2821 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
     2881 taaaacgacg gccagtgcca agcttgcatg cctgcaggtc gactctagag gatccccggg
     2941 taccgagctc gaattcttat cagatcccat gggtatgtat atctccttct taaagttaaa
     3001 caaaattatt tctagcccaa aaaaacgggt atggagaaac agtagagagt tgcgataaaa
     3061 agcgtcaggt aggatccgct aatcttatgg ataaaaatgc tatggcatag caaagtgtga
     3121 cgccgtgcaa ataatcaatg tggacttttc tgccgtgatt atagacactt ttgttacgcg
     3181 tttttgtcat ggctttggtc ccgctttgtt acagaatgct tttaataagc ggggttaccg
     3241 gtttggttag cgagaagagc cagtaaaaga cgcagtgacg gcaatgtctg atgcaatatg
     3301 gacaattggt ttcttctctg aatggcggga gtatgaaaag tatggctgaa gcgcaaaatg
     3361 atcccctgct gccgggatac tcgtttaatg cccatctggt ggcgggttta acgccgattg
     3421 aggccaacgg ttatctcgat ttttttatcg accgaccgct gggaatgaaa ggttatattc
     3481 tcaatctcac cattcgcggt cagggggtgg tgaaaaatca gggacgagaa tttgtttgcc
     3541 gaccgggtga tattttgctg ttcccgccag gagagattca tcactacggt cgtcatccgg
     3601 aggctcgcga atggtatcac cagtgggttt actttcgtcc gcgcgcctac tggcatgaat
     3661 ggcttaactg gccgtcaata tttgccaata cggggttctt tcgcccggat gaagcgcacc
     3721 agccgcattt cagcgacctg tttgggcaaa tcattaacgc cgggcaaggg gaagggcgct
     3781 attcggagct gctggcgata aatctgcttg agcaattgtt actgcggcgc atggaagcga
     3841 ttaacgagtc gctccatcca ccgatggata atcgggtacg cgaggcttgt cagtacatca
     3901 gcgatcacct ggcagacagc aattttgata tcgccagcgt cgcacagcat gtttgcttgt
     3961 cgccgtcgcg tctgtcacat cttttccgcc agcagttagg gattagcgtc ttaagctggc
     4021 gcgaggacca acgtatcagc caggcgaagc tgcttttgag caccacccgg atgcctatcg
     4081 ccaccgtcgg tcgcaatgtt ggttttgacg atcaactcta tttctcgcgg gtatttaaaa
     4141 aatgcaccgg ggccagcccg agcgagttcc gtgccggttg tgaagaaaaa gtgaatgatg
     4201 tagccgtcaa gttgtcataa tttggggatt ccttaaggta tactttccgc tgcataaccc
     4261 tgcttcgggg tcattatagc gattttttcg gtatatccat cctttttcgc acgatataca
     4321 ggattttgcc aaagggttcg tgtagacttt ccttggtgta tccaacggcg tcagccgggc
     4381 aggataggtg aagtaggccc acccgcgagc gggtgttcct tcttcactgt cccttattcg
     4441 cacctggcgg tgctcaacgg gaatcctgct ctgcgaggct ggccgataag ctagcttgtg
     4501 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     4561 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     4621 cccgacagga ctataaagat accaggcgtt tccccctgga ggctccctcg tgcgctctcc
     4681 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     4741 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     4801 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     4861 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     4921 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     4981 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     5041 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     5101 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     5161 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggt
//
