LOCUS       40924_26301        7075 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Integration expression vector pJM252, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7075)
  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 7075)
  AUTHORS   Meisner J.
  TITLE     Direct Submission
  JOURNAL   Submitted (26-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 7075)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7075)
  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 
            (26-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"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..7075
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        827..845
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     CDS             complement(905..1984)
                     /codon_start=1
                     /label=lacI
                     /note="lac repressor"
                     /translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     promoter        complement(1985..2062)
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the
                     promoter boosts expression of the lacI gene about 10-fold."
     promoter        2292..2320
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     protein_bind    2328..2344
                     /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     complement(2397..2413)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(2439..2457)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    complement(2631..2678)
                     /label=FRT
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     oriT            complement(2885..2993)
                     /direction=LEFT
                     /label=oriT
                     /note="incP origin of transfer"
     rep_origin      complement(4418..5006)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5831..7018)
                     /codon_start=1
                     /label=TcR
                     /note="tetracycline efflux protein"
                     /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
                     GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
                     VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
                     LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
                     QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
                     AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
                     AALTSLTSITGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
     promoter        complement(join(7066..7075,1..19))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein
                     gene"
ORIGIN
        1 atcgatgata agctgtcaaa catgagaatt aattcatgtt ctttcctgcg ttatcccctg
       61 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
      121 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc
      181 ctctccccgc gcgttggccg attcattaat gcagcgaatt agcttcaaaa gcgctctgaa
      241 gttcctatac tttctagaga ataggaactt cggaatagga acttcaagat cccctgattc
      301 cctttgtcaa cagcaatgga ttttcaatgg gtcaacatga aatccggctg ttgccctggg
      361 acggcaggtt tgacggcagg cttccaggct ctctcgcaca cgctgggaac actctgggaa
      421 caaacgcgag tgaaaatgga cagtagggcc caggaataat gcggcctcca gcgaggttgg
      481 ggcgtttcat atgagttcga atctcaccgc ctccgccata taacctgctg ttttcattga
      541 tgtttccatt cgacccactc tcaggagtga acgatttggg aacaaactgg gaacgaggcg
      601 cagaaaaaag gggccccgca gggccccttt ttcgttgctt caggaaagtt tcagcgcctg
      661 attcagcaga cctacgatgt cggggccgtc cttgctgaat tagctttatg cttgtaaacc
      721 gttttgtgaa aaaattttta aaataaaaaa ggggacctct agggtcccca attaattagt
      781 aatataatct attaaaggtc attcaaaagg tcatccaccg gcgcgcaatt aaccctcact
      841 aaagggaaca aaagctggag ctccttgcaa ttcgcgctaa cttacattaa ttgcgttgcg
      901 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
      961 acgcgcgggg agaggcggtt tgcgtattgg gcgccagggt ggtttttctt ttcaccagtg
     1021 agacgggcaa cagctgattg cccttcaccg cctggccctg agagagttgc agcaagcggt
     1081 ccacgctggt ttgccccagc aggcgaaaat cctgtttgat ggtggttaac ggcgggatat
     1141 aacatgagct gtcttcggta tcgtcgtatc ccactaccga gatatccgca ccaacgcgca
     1201 gcccggactc ggtaatggcg cgcattgcgc ccagcgccat ctgatcgttg gcaaccagca
     1261 tcgcagtggg aacgatgccc tcattcagca tttgcatggt ttgttgaaaa ccggacatgg
     1321 cactccagtc gccttcccgt tccgctatcg gctgaatttg attgcgagtg agatatttat
     1381 gccagccagc cagacgcaga cgcgccgaga cagaacttaa tgggcccgct aacagcgcga
     1441 tttgctggtg acccaatgcg accagatgct ccacgcccag tcgcgtaccg tcttcatggg
     1501 agaaaataat actgttgatg ggtgtctggt cagagacatc aagaaataac gccggaacat
     1561 tagtgcaggc agcttccaca gcaatggcat cctggtcatc cagcggatag ttaatgatca
     1621 gcccactgac gcgttgcgcg agaagattgt gcaccgccgc tttacaggct tcgacgccgc
     1681 ttcgttctac catcgacacc accacgctgg cacccagttg atcggcgcga gatttaatcg
     1741 ccgcgacaat ttgcgacggc gcgtgcaggg ccagactgga ggtggcaacg ccaatcagca
     1801 acgactgttt gcccgccagt tgttgtgcca cgcggttggg aatgtaattc agctccgcca
     1861 tcgccgcttc cactttttcc cgcgttttcg cagaaacgtg gctggcctgg ttcaccacgc
     1921 gggaaacggt ctgataagag acaccggcat actctgcgac atcgtataac gttactggtt
     1981 tcacattcac caccctgaat tgactctctt ccgggcgcta tcatgccata ccgcgaaagg
     2041 ttttgcacca ttcgatggtg tcaacgtaaa tgccgcttcg ccttcgcgcg cgaattgcaa
     2101 gctgatccgg gcttatcgac tgcacggtgc accaatgctt ctggcgtcag gcagccatcg
     2161 gaagctgtgg tatggctgtg caggtcgtaa atcactgcat aattcgtgtc gctcaaggcg
     2221 cactcccgtt ctggataatg ttttttgcgc cgacatcata acggttctgg caaatattct
     2281 gaaatgagct gttgacaatt aatcatcggc tcgtataatg tgtggaattg tgagcggata
     2341 acaatttcac acaggaaaca gactagtgct ctgcaggaat tcctcgagaa gcttatcgat
     2401 accgtcgacc tcgagggggg gcccggtacc caattcgccc tatagtgagt cgtattacgc
     2461 gcgccggtgg atgacctttt gaatgacctt taatagatta tattactaat taattgggga
     2521 ccctagaggt cccctttttt attttaaaaa ttttttcaca aaacggttta caagcataaa
     2581 gctaattcca ttgagtaagt ttttaagcac atcagcttca aaagcgctct gaagttccta
     2641 tactttctag agaataggaa cttcggaata ggaacttcaa gatccccaat tcgctgcatt
     2701 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     2761 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     2821 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgctagagc
     2881 ttatcggcca gcctcgcaga gcaggattcc cgttgagcac cgccaggtgc gaataaggga
     2941 cagtgaagaa ggaacacccg ctcgcgggtg ggcctacttc acctatcctg cccggctgac
     3001 gccgttggat acaccaagga aagtctacac gaaccctttg gcaaaatcct gtatatcgtg
     3061 cgaaaaagga tggatatacc gaaaaaatcg ctataatgac cccgaagcag ggttatgcag
     3121 cggaaagtat accttaacct ttttcgttgc ttcaggaaag tttcagcgcc tgattcagca
     3181 gacctacgat gtcggggccg tccttgctga tccacttggc gtaatgcttg aagatcatcg
     3241 ccgtcgaggt gtgccccatc tggtccgcga tccattcggg agtggcgatg ccgctgctga
     3301 gcatctggct ggcgaaggtg tggcggcact gattgggacc acgcgagcgg acgcccgcat
     3361 tcctgaggtg gccatgccac cagccgttgc gcagcgtatc cgacgtggag tacgccgccc
     3421 cgctagcact gttgtggaac acgaagcgta cccgttgagg ctttctggtc cggttgtcgc
     3481 ggtctatcac ctcgatctgt acgggcggaa ggcgtctggt tagcttggcc tgctgctgga
     3541 gtgcgcgcag cgccggggcc agtagctgga ccttcctcgt cgatcggcga gtcttggtga
     3601 ccttgtactg gctgcgtaca cgagcgcggc ggaaaaccac ggtccccgtg tccaaatcga
     3661 catcttccca ggccagagct atggcctcgc tgacgcgggg gccgctccag atcatgaact
     3721 ccgccaggtt cagctcaccg atgcgcgcgg tttcagtacc gaggatgagg tcgatttcct
     3781 cacgagtgaa tgggtctggg tcatccgcat ccggaagagt gatcacgatg ccgtctgttg
     3841 ggtcgtgcgc gctccggttc cgagttcgat acagccggaa gatctgccgc aggttgctga
     3901 cgatctctcg caccgtcttg ttgtgcagct tgggcatgag cgtgttctgc acccagtctt
     3961 gaatatccag gtggtcgatg ctgtcggcct ggctgtcacc ccagcgaggt cggatgtggg
     4021 tttccacgcg gctggtgtag ccgcggaagc cgctggcggc aatctggttg cgtttgatgt
     4081 cgagccagag gtcgatgtaa tggcccaagg tgttgctttt cacccgcggt gagtccggga
     4141 agtgtcggct gtagctgaag actccttgct tgatctcgta gttgatgatg ccggccagcc
     4201 gttcggcatt ggctatattc tcgggcgttg cgtcccctgg tatggactcc ctacacagct
     4261 cgccctggta gcggaagtag atcctgatcc gttttccgcg tacctcgact ccgtctgcca
     4321 tctggttccc ctgatccaca ccaagcgttc aagctggcat gtgagcaaaa ggccagcaaa
     4381 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     4441 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     4501 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     4561 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     4621 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     4681 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     4741 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     4801 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga
     4861 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     4921 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     4981 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     5041 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     5101 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     5161 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     5221 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     5281 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     5341 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     5401 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     5461 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     5521 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     5581 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     5641 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     5701 ccgtaagatg cttttctgtg actggtgagt caagggttgg tttgcgcatt cacagttctc
     5761 cgcaagaatt gattggctcc aattcttgga gtggtgaatc cgttagcgag gtgccgccgg
     5821 cttccattca ggtcgaggtg gcccggctcc atgcaccgcg acgcaacgcg gggaggcaga
     5881 caaggtatag ggcggcgcct acaatccatg ccaacccgtt ccatgtgctc gccgaggcgg
     5941 cataaatcgc cgtgacgatc agcggtccag tgatcgaagt taggctggta agagccgcga
     6001 gcgatccttg aagctgtccc tgatggtcgt catctacctg cctggacagc atggcctgca
     6061 acgcgggcat cccgatgccg ccggaagcga gaagaatcat aatggggaag gccatccagc
     6121 ctcgcgtcgc gaacgccagc aagacgtagc ccagcgcgtc ggccgccatg ccggcgataa
     6181 tggcctgctt ctcgccgaaa cgtttggtgg cgggaccagt gacgaaggct tgagcgaggg
     6241 cgtgcaagat tccgaatacc gcaagcgaca ggccgatcat cgtcgcgctc cagcgaaagc
     6301 ggtcctcgcc gaaaatgacc cagagcgctg ccggcacctg tcctacgagt tgcatgataa
     6361 agaagacagt cataagtgcg gcgacgatag tcatgccccg cgcccaccgg aaggagctga
     6421 ctgggttgaa ggctctcaag ggcatcggtc ggcgctctcc cttatgcgac tcctgcatta
     6481 ggaagcagcc cagtagtagg ttgaggccgt tgagcaccgc cgccgcaagg aatggtgcat
     6541 gtaaggagat ggcgcccaac agtcccccgg ccacggggcc tgccaccata cccacgccga
     6601 aacaagcgct catgagcccg aagtggcgag cccgatcttc cccatcggtg atgtcggcga
     6661 tataggcgcc agcaaccgca cctgtggcgc cggtgatgcc ggccacgatg cgtccggcgt
     6721 agagaatcca caggacgggt gtggtcgcca tgatcgcgta gtcgatagtg gctccaagta
     6781 gcgaagcgag caggactggg cggcggccaa agcggtcgga cagtgctccg agaacgggtg
     6841 cgcatagaaa ttgcatcaac gcatatagcg ctagcagcac gccatagtga ctggcgatgc
     6901 tgtcggaatg gacaatatcc cgcaagaggc ccggcagtac cggcataacc aagcctatgc
     6961 ctacagcatc cagggtgacg gtgccgagga tgacgatgag cgcattgtta gatttcatac
     7021 acggtgcctg actgcgttag caatttaact gtgataaact accgcattaa agcta
//
