LOCUS       40924_26291        6694 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Integration expression vector pJM220, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6694)
  AUTHORS   Meisner J, Goldberg JB.
  TITLE     Escherichia coli rhaSR-PrhaBAD inducible promoter system allows 
            tightly controlled gene expression over a wide range in Pseudomonas 
            aeruginosa
  JOURNAL   Appl. Environ. Microbiol. (2016) In press
  PUBMED    27613678
REFERENCE   2  (bases 1 to 6694)
  AUTHORS   Meisner J.
  TITLE     Direct Submission
  JOURNAL   Submitted (25-AUG-2016) Biochemistry, Emory University School of 
            Medicine, 1510 Clifton Road NE, Rollins Research Center, room G223, 
            Atlanta, GA 30322, USA
REFERENCE   3  (bases 1 to 6694)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6694)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Appl. 
            Environ. Microbiol. (2016) In press"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (25-AUG-2016) Biochemistry, Emory University School of Medicine, 
            1510 Clifton Road NE, Rollins Research Center, room G223, Atlanta, 
            GA 30322, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6694
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     mobile_element  382..580
                     /mobile_element_type="transposon:Tn7R"
                     /label=n7R
                     /note="right end of Tn7"
     primer_bind     585..601
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      629..723
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     terminator      826..912
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     protein_bind    958..1005
                     /label=FRT
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             complement(1138..1668)
                     /label=GmR
                     /note="gentamycin acetyltransferase"
     promoter        complement(1857..1885)
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     misc_feature    1926..1973
                     /note="FRT; Flp recombinase target site"
     protein_bind    1926..1973
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             complement(2116..2961)
                     /label=rhaR
                     /note="transcriptional activator of rhaR and rhaS"
     CDS             complement(3038..3871)
                     /label=rhaS
                     /note="positive regulator of the rhaB promoter"
     regulatory      complement(3896..3931)
                     /label=Escherichia coli rhaSR promoter
                     /note="Escherichia coli rhaSR promoter"
                     /regulatory_class="promoter"
     misc_feature    3928..3944
                     /label=rhaI4 operator
                     /note="rhaI4 operator"
     misc_feature    3962..3978
                     /label=rhaI3 operator
                     /note="rhaI3 operator"
     promoter        4017..4135
                     /label=rhaB promoter
                     /note="promoter of the E. coli rhaBAD operon, conferring
                     tight induction with L-rhamnose and repression with 
                     D-glucose in the presence of RhaR and RhaS (Giacalone et 
                     al., 2006)"
     misc_feature    4146..4202
                     /note="MCS; multiple cloning site"
     mobile_element  complement(4211..4376)
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     oriT            complement(4535..4643)
                     /direction=LEFT
                     /label=oriT
                     /note="incP origin of transfer"
     rep_origin      complement(4875..5463)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5637..6494)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(6495..6599)
                     /label=AmpR promoter
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct agaggaccag ccgcgtaacc tggcaaaatc ggttacggtt gagtaataaa
      361 tggatgccct gcgtaagcgg gtgtgggcgg acaataaagt cttaaactga acaaaataga
      421 tctaaactat gacaataaag tcttaaacta gacagaatag ttgtaaactg aaatcagtcc
      481 agttatgctg tgaaaaagca tactggactt ttgttatggc taaagcaaac tcttcatttt
      541 ctgaagtgca aattgcccgt cgtattaaag aggggcgtgg ggttcgaggt cgacggtatc
      601 gataagctag cttaattagc tgagcttgga ctcctgttga tagatccagt aatgacctca
      661 gaactccatc tggatttgtt cagaacgctc ggttgccgcc gggcgttttt tattggtgag
      721 aatccaagct agactgcgat gagtggcagg gcggggcgta atttttttaa ggcagttatt
      781 ggtgccctta aacgcctggg gtaatgactc tctagcttga ggcatcaaat aaaacgaaag
      841 gctcagtcga aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctcctg
      901 agtaggacaa atccgccgct aggagcttgc ggcccggacg atcgaattgg ggatcttgaa
      961 gttcctattc cgaagttcct attctctaga aagtatagga acttcagagc gcttttgaag
     1021 ctgatgtgct taaaaactta ctcaatggaa ttctagatcg aattggccgc ggcgttgtga
     1081 caatttaccg aacaactccg cggccgggaa gccgatctcg gcttgaacga attgttaggt
     1141 ggcggtactt gggtcgatat caaagtgcat cacttcttcc cgtatgccca actttgtata
     1201 gagagccact gcgggatcgt caccgtaatc tgcttgcacg tagatcacat aagcaccaag
     1261 cgcgttggcc tcatgcttga ggagattgat gagcgcggtg gcaatgccct gcctccggtg
     1321 ctcgccggag actgcgagat catagatata gatctcacta cgcggctgct caaacctggg
     1381 cagaacgtaa gccgcgagag cgccaacaac cgcttcttgg tcgaaggcag caagcgcgat
     1441 gaatgtctta ctacggagca agttcccgag gtaatcggag tccggctgat gttgggagta
     1501 ggtggctacg tctccgaact cacgaccgaa aagatcaaga gcagcccgca tggatttgac
     1561 ttggtcaggg ccgagcctac atgtgcgaat gatgcccata cttgagccac ctaactttgt
     1621 tttagggcga ctgccctgct gcgtaacatc gttgctgctg cgtaacatcg ttgctgctcc
     1681 ataacatcaa acatcgaccc acggcgtaac gcgcttgctg cttggatgcc cgaggcatag
     1741 actgtacaaa aaaacagtca taacaagcca tgaaaaccgc cactgcgccg ttaccaccgc
     1801 tgcgttcggt caaggttctg gaccagttgc gtgagcgcat acgctacttg cattacagtt
     1861 tacgaaccga acaggcttat gtcaattcga tccattgctg ttgacaaagg gaatcagggg
     1921 atcttgaagt tcctattccg aagttcctat tctctagaaa gtataggaac ttcagagcgc
     1981 ttttgaagct aattcgatca tgcatgagct caaagagtgg aacaatgcag gacgccgtcc
     2041 ggtaacggtg ttgagcctcg gttgtgtggt gattattgtc gccgctaaca tcgtcggcat
     2101 cggcatggcg aattaatctt tctgcgaatt gagatgacgc cactggctgg gcgtcatccc
     2161 ggtttcccgg gtaaacacca ccgaaaaata gttactatct tcaaagccac attcggtcga
     2221 aatatcactg attaacaggc ggctatgctg gagaagatat tgcgcatgac acactctgac
     2281 ctgtcgcaga tattgattga tggtcattcc agtctgctgg cgaaattgct gacgcaaaac
     2341 gcgctcactg cacgatgcct catcacaaaa tttatccagc gcaaagggac ttttcaggct
     2401 agccgccagc cgggtaatca gcttatccag caacgtttcg ctggatgttg gcggcaacga
     2461 atcactggtg taacgatggc gattcagcaa catcaccaac tgcccgaaca gcaactcagc
     2521 catttcgtta gcaaacggca catgctgact actttcatgc tcaagctgac cgataacctg
     2581 ccgcgcctgc gccatcccca tgctacctaa gcgccagtgt ggttgccctg cgctggcgtt
     2641 aaatcccgga atcgccccct gccagtcaag attcagcttc agacgctccg ggcaataaat
     2701 aatattctgc aaaaccagat cgttaacgga agcgtaggag tgtttatcgt cagcatgaat
     2761 gtaaaagaga tcgccacggg taatgcgata agggcgatcg ttgagtacat gcaggccatt
     2821 accgcgccag acaatcacca gctcacaaaa atcatgtgta tgttcagcaa agacatcttg
     2881 cggataacgg tcagccacag cgactgcctg ctggtcgctg gcaaaaaaat catctttgag
     2941 aagttttaac tgatgcgcca ccgtggctac ctcggccaga gaacgaagtt gattattcgc
     3001 aatatggcgt acaaatacgt tgagaagatt cgcgttattg cagaaagcca tcccgtccct
     3061 ggcgaatatc acgcggtgac cagttaaact ctcggcgaaa aagcgtcgaa aagtggttac
     3121 tgtcgctgaa tccacagcga taggcgatgt cagtaacgct ggcctcgctg tggcgtagca
     3181 gatgtcgggc tttcatcagt cgcaggcggt tcaggtatcg ctgaggcgtc agtcccgttt
     3241 gctgcttaag ctgccgatgt agcgtacgca gtgaaagaga aaattgatcc gccacggcat
     3301 cccaattcac ctcatcggca aaatggtcct ccagccaggc cagaagcaag ttgagacgtg
     3361 atgcgctgtt ttccaggttc tcctgcaaac tgcttttacg cagcaagagc agtaattgca
     3421 taaacaagat ctcgcgactg gcggtcgagg gtaaatcatt ttccccttcc tgctgttcca
     3481 tctgtgcaac cagctgtcgc acctgctgca atacgctgtg gttaacgcgc cagtgagacg
     3541 gatactgccc atccagctct tgtggcagca actgattcag cccggcgaga aactgaaatc
     3601 gatccggcga gcgatacagc acattggtca gacacagatt atcggtatgt tcatacagat
     3661 gccgatcatg atcgcgtacg aaacagaccg tgccaccggt gatggtatag ggctgcccat
     3721 taaacacatg aatacccgtg ccatgttcga caatcacaat ttcatgaaaa tcatgatgat
     3781 gttcaggaaa atccgcctgc gggagccggg gttctatcgc cacggacgcg ttaccagacg
     3841 gaaaaaaatc cacactatgt aatacggtca tactggcctc ctgatgtcgt caacacggcg
     3901 aaatagtaat cacgaggtca ggttcttacc ttaaattttc gacggaaaac cacgtaaaaa
     3961 acgtcgattt ttcaagatac agcgtgaatt ttcaggaaat gcggtgagca tcacatcacc
     4021 acaattcagc aaattgtgaa catcatcacg ttcatctttc cctggttgcc aatggcccat
     4081 tttcctgtca gtaacgagaa ggtcgcgaat tcaggcgctt tttagactgg tcgtaatgaa
     4141 attcaactag tgctctgcag gaattcctcg agaagcttgg gcccggtacc tcgcgaaggc
     4201 cttgcaggcc aaccagataa gtgaaatcta gttccaaact attttgtcat ttttaatttt
     4261 cgtattagct tacgacgcta cacccagttc ccatctattt tgtcactctt ccctaaataa
     4321 tccttaaaaa ctccatttcc acccctccca gttcccaact attttgtccg cccacagcgg
     4381 ggcatttttc ttcctgttat gtttgggcgc tgcattaatg aatcggccaa cgcgcgggga
     4441 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgacccg ctgcgctcgg
     4501 tcgttcggct gcggcgagcg gtatcagagc ttatcggcca gcctcgcaga gcaggattcc
     4561 cgttgagcac cgccaggtgc gaataaggga cagtgaagaa ggaacacccg ctcgcgggtg
     4621 ggcctacttc acctatcctg cccggctgac gccgttggat acaccaagga aagtctacac
     4681 gaaccctttg gcaaaatcct gtatatcgtg cgaaaaagga tggatatacc gaaaaaatcg
     4741 ctataatgac cccgaagcag ggttatgcag cggaaagtat accttaagga atcccctgat
     4801 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     4861 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     4921 tcgagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     4981 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     5041 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     5101 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     5161 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     5221 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     5281 ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg
     5341 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     5401 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     5461 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     5521 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     5581 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     5641 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     5701 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     5761 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     5821 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     5881 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     5941 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     6001 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     6061 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     6121 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     6181 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     6241 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     6301 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     6361 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     6421 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     6481 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     6541 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     6601 acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc
     6661 tataaaaata ggcgtatcac gaggcccttt cgtc
//
