LOCUS       40924_24478        6166 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Escherichia-Pseudomonas shuttle vector pHERD26T, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6166)
  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 6166)
  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 6166)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6166)
  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..6166
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(46..397)
                     /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             411..1241
                     /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     1405..1421
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    complement(1425..1481)
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     RBS             complement(1507..1529)
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     promoter        complement(1556..1840)
                     /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             1867..2742
                     /codon_start=1
                     /label=araC
                     /note="L-arabinose regulatory protein"
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     oriT            2902..3010
                     /label=oriT
                     /note="incP origin of transfer"
     rep_origin      complement(3082..3670)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4495..5682)
                     /codon_start=1
                     /label=TcR
                     /note="tetracycline efflux protein"
                     /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
                     GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
                     VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
                     LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
                     QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
                     AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
                     AALTSLTSITGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
     promoter        complement(5730..5758)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein
                     gene"
     protein_bind    6005..6026
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        6041..6071
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6079..6095
                     /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)."
     primer_bind     6103..6119
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        6137..6155
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
ORIGIN
        1 cctctcaggc gccgctggtg ccgctggttg gacgccaagg gtgaatccgc ctcgataccc
       61 tgattactcg cttcctgcgc cctctcaggc ggcgataggg gactggtaaa acggggattg
      121 cccagacgcc tcccccgccc cttcaggggc acaaatgcgg ccccaacggg gccacgtagt
      181 ggtgcgtttt ttgcgtttcc acccttttct tccttttccc ttttaaacct tttaggacgt
      241 ctacaggcca cgtaatccgt ggcctgtaga gtttaaaaag ggacggattt gttgccatta
      301 agggacggat ttgttgttaa gaagggacgg atttgttgtt gtaaagggac ggatttgttg
      361 tattgtggga cgcagataca gtgtcccctt atacacaagg aatgtcgaac gtggcctcac
      421 ccccaatggt ttacaaaagc aatgccctgg tcgaggccgc gtatcgcctc agtgttcagg
      481 aacagcggat cgttctggcc tgtattagcc aggtgaagag gagcgagcct gtcaccgatg
      541 aagtgatgta ttcagtgacg gcggaggaca tagcgacgat ggcgggtgtc cctatcgaat
      601 cttcctacaa ccagctcaaa gaagcggccc tgcgcctgaa acggcgggaa gtccggttaa
      661 cccaagagcc caatggcaag gggaaaagac cgagtgtgat gattaccggc tgggtgcaaa
      721 caatcatcta ccgggagggt gagggccgtg tagaactcag gttcaccaaa gacatgctgc
      781 cgtacctgac ggaactcacc aaacagttca ccaaatacgc cttggctgac gtggccaaga
      841 tggacagcac ccacgcgatc aggctttacg agctgctcat gcaatgggac agcatcggcc
      901 agcgcgaaat agaaattgac cagctgcgaa agtggtttca actggaaggc cggtatccct
      961 cgatcaagga cttcaagttg cgagtgcttg atccagccgt gacgcagatc aacgagcaca
     1021 gcccgctaca ggtggagtgg gcgcagcgaa agaccgggcg caaggtcaca catctgttgt
     1081 tcagttttgg accgaagaag cccgccaagg cggtgggtaa ggccccagcg aagcgcaagg
     1141 ccgggaagat ttcagatgct gagatcgcga aacaggctcg ccctggtgag acatgggaag
     1201 cggcccgcgc tcgactaacc cagatgccgc tggatctggc ctagaggccg tggccaccac
     1261 ggcccggcct gcctttcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt
     1321 cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc
     1381 cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgccaagctt gcatgcctgc
     1441 aggtcgactc tagaggatcc ccgggtaccg agctcgaatt cttatcagat cccatgggta
     1501 tgtatatctc cttcttaaag ttaaacaaaa ttatttctag cccaaaaaaa cgggtatgga
     1561 gaaacagtag agagttgcga taaaaagcgt caggtaggat ccgctaatct tatggataaa
     1621 aatgctatgg catagcaaag tgtgacgccg tgcaaataat caatgtggac ttttctgccg
     1681 tgattataga cacttttgtt acgcgttttt gtcatggctt tggtcccgct ttgttacaga
     1741 atgcttttaa taagcggggt taccggtttg gttagcgaga agagccagta aaagacgcag
     1801 tgacggcaat gtctgatgca atatggacaa ttggtttctt ctctgaatgg cgggagtatg
     1861 aaaagtatgg ctgaagcgca aaatgatccc ctgctgccgg gatactcgtt taatgcccat
     1921 ctggtggcgg gtttaacgcc gattgaggcc aacggttatc tcgatttttt tatcgaccga
     1981 ccgctgggaa tgaaaggtta tattctcaat ctcaccattc gcggtcaggg ggtggtgaaa
     2041 aatcagggac gagaatttgt ttgccgaccg ggtgatattt tgctgttccc gccaggagag
     2101 attcatcact acggtcgtca tccggaggct cgcgaatggt atcaccagtg ggtttacttt
     2161 cgtccgcgcg cctactggca tgaatggctt aactggccgt caatatttgc caatacgggg
     2221 ttctttcgcc cggatgaagc gcaccagccg catttcagcg acctgtttgg gcaaatcatt
     2281 aacgccgggc aaggggaagg gcgctattcg gagctgctgg cgataaatct gcttgagcaa
     2341 ttgttactgc ggcgcatgga agcgattaac gagtcgctcc atccaccgat ggataatcgg
     2401 gtacgcgagg cttgtcagta catcagcgat cacctggcag acagcaattt tgatatcgcc
     2461 agcgtcgcac agcatgtttg cttgtcgccg tcgcgtctgt cacatctttt ccgccagcag
     2521 ttagggatta gcgtcttaag ctggcgcgag gaccaacgta tcagccaggc gaagctgctt
     2581 ttgagcacca cccggatgcc tatcgccacc gtcggtcgca atgttggttt tgacgatcaa
     2641 ctctatttct cgcgggtatt taaaaaatgc accggggcca gcccgagcga gttccgtgcc
     2701 ggttgtgaag aaaaagtgaa tgatgtagcc gtcaagttgt cataatttgg ggattcctta
     2761 aggtatactt tccgctgcat aaccctgctt cggggtcatt atagcgattt tttcggtata
     2821 tccatccttt ttcgcacgat atacaggatt ttgccaaagg gttcgtgtag actttccttg
     2881 gtgtatccaa cggcgtcagc cgggcaggat aggtgaagta ggcccacccg cgagcgggtg
     2941 ttccttcttc actgtccctt attcgcacct ggcggtgctc aacgggaatc ctgctctgcg
     3001 aggctggccg ataagctagc ttatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     3061 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     3121 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     3181 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     3241 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     3301 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     3361 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     3421 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     3481 agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg
     3541 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     3601 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     3661 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     3721 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     3781 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     3841 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     3901 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     3961 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     4021 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     4081 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     4141 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     4201 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     4261 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     4321 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     4381 tgtgactggt gagtcaaggg ttggtttgcg cattcacagt tctccgcaag aattgattgg
     4441 ctccaattct tggagtggtg aatccgttag cgaggtgccg ccggcttcca ttcaggtcga
     4501 ggtggcccgg ctccatgcac cgcgacgcaa cgcggggagg cagacaaggt atagggcggc
     4561 gcctacaatc catgccaacc cgttccatgt gctcgccgag gcggcataaa tcgccgtgac
     4621 gatcagcggt ccagtgatcg aagttaggct ggtaagagcc gcgagcgatc cttgaagctg
     4681 tccctgatgg tcgtcatcta cctgcctgga cagcatggcc tgcaacgcgg gcatcccgat
     4741 gccgccggaa gcgagaagaa tcataatggg gaaggccatc cagcctcgcg tcgcgaacgc
     4801 cagcaagacg tagcccagcg cgtcggccgc catgccggcg ataatggcct gcttctcgcc
     4861 gaaacgtttg gtggcgggac cagtgacgaa ggcttgagcg agggcgtgca agattccgaa
     4921 taccgcaagc gacaggccga tcatcgtcgc gctccagcga aagcggtcct cgccgaaaat
     4981 gacccagagc gctgccggca cctgtcctac gagttgcatg ataaagaaga cagtcataag
     5041 tgcggcgacg atagtcatgc cccgcgccca ccggaaggag ctgactgggt tgaaggctct
     5101 caagggcatc ggtcggcgct ctcccttatg cgactcctgc attaggaagc agcccagtag
     5161 taggttgagg ccgttgagca ccgccgccgc aaggaatggt gcatgtaagg agatggcgcc
     5221 caacagtccc ccggccacgg ggcctgccac catacccacg ccgaaacaag cgctcatgag
     5281 cccgaagtgg cgagcccgat cttccccatc ggtgatgtcg gcgatatagg cgccagcaac
     5341 cgcacctgtg gcgccggtga tgccggccac gatgcgtccg gcgtagagaa tccacaggac
     5401 gggtgtggtc gccatgatcg cgtagtcgat agtggctcca agtagcgaag cgagcaggac
     5461 tgggcggcgg ccaaagcggt cggacagtgc tccgagaacg ggtgcgcata gaaattgcat
     5521 caacgcatat agcgctagca gcacgccata gtgactggcg atgctgtcgg aatggacgat
     5581 atcccgcaag aggcccggca gtaccggcat aaccaagcct atgcctacag catccagggt
     5641 gacggtgccg aggatgacga tgagcgcatt gttagatttc atacacggtg cctgactgcg
     5701 ttagcaattt aactgtgata aactaccgca ttaaagctaa tcgatgataa gctgtcaaac
     5761 atgagaatta attcatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     5821 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     5881 tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga
     5941 ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg
     6001 caattaatgt gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg
     6061 ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc
     6121 atgattacgc caagctattt aggtgacact atagaatact caagct
//
