LOCUS       sequence-235414-        7079 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Integration vector for Pseudomonas aeruginosa with lacIq-Ptac 
            inducible promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7079)
  AUTHORS   Meisner J, Goldberg JB
  TITLE     The Escherichia coli rhaSR-PrhaBAD Inducible Promoter System Allows 
            Tightly Controlled Gene Expression over a Wide Range in Pseudomonas 
            aeruginosa.
  JOURNAL   Appl Environ Microbiol. 2016 Oct 27;82(22):6715-6727. doi: 
            10.1128/AEM.02041-16. Print 2016 Nov 15.
  PUBMED    27613678
REFERENCE   2  (bases 1 to 7079)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1128/AEM.02041-16"; journalName: "Appl Environ Microbiol"; date:
            "2016-10-27- 27"; volume: "82"; issue: "22"; pages: "6715-6727"
COMMENT     Sequence Label: pJM252 (miniCTX1-lacIq-Ptac)
FEATURES             Location/Qualifiers
     source          1..7079
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(238..285)
                     /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)."
     primer_bind     822..842
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        824..842
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     CDS             complement(899..1981)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     1943..1962
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(1982..2059)
                     /gene="lacI (mutant)"
                     /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        join(2289..2294,2295..2310,2311..2317)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    2325..2341
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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     2330..2352
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     complement(2394..2410)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2395..2411)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(2435..2454)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2436..2454)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    complement(2628..2675)
                     /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)."
     oriT            2882..2990
                     /note="incP origin of transfer"
     rep_origin      complement(4422..5010)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4502..4521)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5832..7022)
                     /codon_start=1
                     /gene="tet"
                     /product="tetracycline efflux protein"
                     /label=TcR
                     /note="confers resistance to tetracycline"
                     /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
                     GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
                     VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
                     LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
                     QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
                     AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
                     AALTSLTSIIGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
     primer_bind     6651..6670
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        complement(join(7070..7075,7076..7079,1..13,14..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 tttcaatggg tcaacatgaa atccggctgt tgccctggga
      361 cggcaggttt gacggcaggc ttccaggctc tctcgcacac gctgggaaca ctctgggaac
      421 aaacgcgagt gaaaatggac agtagggccc aggaataatg cggcctccag cgaggttggg
      481 gcgtttcata tgagttcgaa tctcaccgcc tccgccatat aacctgctgt tttcattgat
      541 gtttccattc gacccactct caggagtgaa cgatttggga acaaactggg aacgaggcgc
      601 agaaaaaagg ggccccgcag ggcccctttt tcgttgcttc aggaaagttt cagcgcctga
      661 ttcagcagac ctacgatgtc ggggccgtcc ttgctgaatt agctttatgc ttgtaaaccg
      721 ttttgtgaaa aaatttttaa aataaaaaag gggacctcta gggtccccaa ttaattagta
      781 atataatcta ttaaaggtca ttcaaaaggt catccacccg cgcaattaac cctcactaaa
      841 gggaacaaaa gctggagctc cttgcaattc gcgctaactt acattaattg cgttgcgctc
      901 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
      961 cgcggggaga ggcggtttgc gtattgggcg ccagggtggt ttttcttttc accagtgaga
     1021 cgggcaacag ctgattgccc ttcaccgcct ggccctgaga gagttgcagc aagcggtcca
     1081 cgctggtttg ccccagcagg cgaaaatcct gtttgatggt ggttaacggc gggatataac
     1141 atgagctgtc ttcggtatcg tcgtatccca ctaccgagat atccgcacca acgcgcagcc
     1201 cggactcggt aatggcgcgc attgcgccca gcgccatctg atcgttggca accagcatcg
     1261 cagtgggaac gatgccctca ttcagcattt gcatggtttg ttgaaaaccg gacatggcac
     1321 tccagtcgcc ttcccgttcc gctatcggct gaatttgatt gcgagtgaga tatttatgcc
     1381 agccagccag acgcagacgc gccgagacag aacttaatgg gcccgctaac agcgcgattt
     1441 gctggtgacc caatgcgacc agatgctcca cgcccagtcg cgtaccgtct tcatgggaga
     1501 aaataatact gttgatgggt gtctggtcag agacatcaag aaataacgcc ggaacattag
     1561 tgcaggcagc ttccacagca atggcatcct ggtcatccag cggatagtta atgatcagcc
     1621 cactgacgcg ttgcgcgaga agattgtgca ccgccgcttt acaggcttcg acgccgcttc
     1681 gttctaccat cgacaccacc acgctggcac ccagttgatc ggcgcgagat ttaatcgccg
     1741 cgacaatttg cgacggcgcg tgcagggcca gactggaggt ggcaacgcca atcagcaacg
     1801 actgtttgcc cgccagttgt tgtgccacgc ggttgggaat gtaattcagc tccgccatcg
     1861 ccgcttccac tttttcccgc gttttcgcag aaacgtggct ggcctggttc accacgcggg
     1921 aaacggtctg ataagagaca ccggcatact ctgcgacatc gtataacgtt actggtttca
     1981 cattcaccac cctgaattga ctctcttccg ggcgctatca tgccataccg cgaaaggttt
     2041 tgcaccattc gatggtgtca acgtaaatgc cgcttcgcct tcgcgcgcga attgcaagct
     2101 gatccgggct tatcgactgc acggtgcacc aatgcttctg gcgtcaggca gccatcggaa
     2161 gctgtggtat ggctgtgcag gtcgtaaatc actgcataat tcgtgtcgct caaggcgcac
     2221 tcccgttctg gataatgttt tttgcgccga catcataacg gttctggcaa atattctgaa
     2281 atgagctgtt gacaattaat catcggctcg tataatgtgt ggaattgtga gcggataaca
     2341 atttcacaca ggaaacagac tagtgctctg caggaattcc tcgagaagct tatcgatacc
     2401 gtcgacctcg agggggggcc cggtacccaa ttcgccctat agtgagtcgt attacgcgcg
     2461 ccggtggatg accttttgaa tgacctttaa tagattatat tactaattaa ttggggaccc
     2521 tagaggtccc cttttttatt ttaaaaattt tttcacaaaa cggtttacaa gcataaagct
     2581 aattccattg agtaagtttt taagcacatc agcttcaaaa gcgctctgaa gttcctatac
     2641 tttctagaga ataggaactt cggaataggt acttcaagat ccccaattcg ctgcattaat
     2701 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     2761 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     2821 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg ctagagctta
     2881 tcggccagcc tcgcagagca ggattcccgt tgagcaccgc caggtgcgaa taagggacag
     2941 tgaagaagga acacccgctc gcgggtgggc ctacttcacc tatcctgccc ggctgacgcc
     3001 gttggataca ccaaggaaag tctacacgaa ccctttggca aaatcctgta tatcgtgcga
     3061 aaaaggatgg atataccgaa aaaatcgcta taatgacccc gaagcagggt tatgcagcgg
     3121 aaagtatacc ttaacctttt tcgttgcttc aggaaagttt cagcgcctga ttcagcagac
     3181 ctacgatgtc ggggccgtcc ttgctgatcc acttggcgta atgcttgaag atcatcgccg
     3241 tcgaggtgtg ccccatctgg tccgcgatcc attcgggagt ggcgatgccg ctgctgagca
     3301 tctggctggc gaaggtgtgg cggcactgat tgggaccacg cgagcggacg cccgcattcc
     3361 tgaggtggcc atgccaccag ccgttgcgca gcgtatccga cgtggagtac gccgccccgc
     3421 tagcactgtt gtggaacacg aagcgtaccc gttgaggctt tctggtccgg ttgtcgcggt
     3481 ctatcacctc gatctgtacg ggcggaaggc gtctggttag cttggcctgc tgctggagtg
     3541 cgcgcagcgc cggggccagt agctggacct tcctcgtcga tcggcgagtc ttggtgacct
     3601 tgtactggct gcgtacacga gcgcggcgga aaaccacggt ccccgtgtcc aaatcgacat
     3661 cttcccaggc cagagctatg gcctcgctga cgcgggggcc gctccagatc atgaactccg
     3721 ccaggttcag ctcaccgatg cgcgcggttt cagtaccgag gatgaggtcg atttcctcac
     3781 gagtgaatgg gtctgggtca tccgcatccg gaagagtgat cacgatgccg tctgttgggt
     3841 cgtgcgcgct ccggttccga gttcgataca gccggaagat ctgccgcagg ttgctgacga
     3901 tctctcgcac cgtcttgttg tgcagcttgg gcatgagcgt gttctgcacc cagtcttgaa
     3961 tatccaggtg gtcgatgctg tcggcctggc tgtcacccca gcgaggtcgg atgtgggttt
     4021 ccacgcggct ggtgtagccg cggaagccgc tggcggcaat ctggttgcgt ttgatgtcga
     4081 gccagaggtc gatgtaatgg cccaaggtgt tgcttttcac ccgcggtgag tccgggaagt
     4141 gtcggctgta gctgaagact ccttgcttga tctcgtagtt gatgatgccg gccagctgtt
     4201 cggcattggc tatattctcg ggcgttgcgt cccctggtat ggactcccta cacagctcgc
     4261 cctggtagcg gaagtagatc ctgatccgtt ttccgcgtac ctcgactccg tctgccatct
     4321 ggttcccctg atccacacca agcgttcagg ctaccaagct ggatgtgagc aaaaggccag
     4381 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     4441 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     4501 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     4561 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     4621 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     4681 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     4741 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     4801 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     4861 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     4921 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     4981 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     5041 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     5101 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     5161 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     5221 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     5281 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     5341 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     5401 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     5461 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     5521 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     5581 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     5641 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     5701 ccatccgtaa gatgcttttc tgtgactggt gagtcaaggg ttggtttgcg cattcacagt
     5761 tctccgcaag aattgattgg ctccaattct tggagtggtg aatccgttag cgaggtgccg
     5821 ccggcttcca ttcaggtcga ggtggcccgg ctccatgcac cgcgacgcaa cgcggggagg
     5881 cagacaaggt atagggcggc gcctacaatc catgccaacc cgttccatgt gctcgccgag
     5941 gcggcataaa tcgccgtgac gatcagcggt ccaatgatcg aagttaggct ggtaagagcc
     6001 gcgagcgatc cttgaagctg tccctgatgg tcgtcatcta cctgcctgga cagcatggcc
     6061 tgcaacgcgg gcatcccgat gccgccggaa gcgagaagaa tcataatggg gaaggccatc
     6121 cagcctcgcg tcgcgaacgc cagcaagacg tagcccagcg cgtcggccgc catgccggcg
     6181 ataatggcct gcttctcgcc gaaacgtttg gtggcgggac cagtgacgaa ggcttgagcg
     6241 agggcgtgca agattccgaa taccgcaagc gacaggccga tcatcgtcgc gctccagcga
     6301 aagcggtcct cgccgaaaat gacccagagc gctgccggca cctgtcctac gagttgcatg
     6361 ataaagaaga cagtcataag tgcggcgacg atagtcatgc cccgcgccca ccggaaggag
     6421 ctgactgggt tgaaggctct caagggcatg ggtcggcgct ctcccttatg cgactcctgc
     6481 attaggaagc agcccagtag taggttgagg ccgttgagca ccgccgccgc aaggaatggt
     6541 gcatgtaagg agatggcgcc caacagtccc ccggccacgg ggcctgccac catacccacg
     6601 ccgaaacaag cgctcatgag cccgaagtgg cgagcccgat cttccccatc ggtgatgtcg
     6661 gcgatatagg cgccagcaac cgcacctgtg gcgccggtga tgccggccac gatgcgtccg
     6721 gcgtagagaa tccacaggac gggtgtggtc gccatgatcg cgtagtcgat agtggctcca
     6781 agtagcgaag cgagcaggac tgggcggcgg ccaaagcggt cggacagtgc tccgagaacg
     6841 ggtgcgcata gaaattgcat caacgcatat agcgctagca gcacgccata gtgactggcg
     6901 atgctgtcgg aatggacaat atcccgcaag aggcccggca gtaccggcat aaccaagcct
     6961 atgcctacag catccagggt gacggtgccg aggatgacga tgagcgcatt gttagatttc
     7021 atacacggtg cctgactgcg ttagcaattt aactgtgata aactaccgca ttaaagcta
//
