LOCUS       sequence-380611-        8573 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Test the biological function of the spTse5-CT (1169-1317)-PhoA 
            (22-474)-LacZ (4-60) fusion protein in P. putida .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8573)
  AUTHORS   Gonzalez-Magana A, Altuna J, Queralt-Martin M, Largo E, Velazquez C,
            Montanchez I, Bernal P, Alcaraz A, Albesa-Jove D
  TITLE     The P. aeruginosa effector Tse5 forms membrane pores disrupting the 
            membrane potential of intoxicated bacteria.
  JOURNAL   Commun Biol. 2022 Nov 5;5(1):1189. doi: 10.1038/s42003-022-04140-y.
  PUBMED    36335275
REFERENCE   2  (bases 1 to 8573)
  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"; journalName: "Commun 
            Biol."; date: "2022-11-5"; volume: "5(1):1189. doi"; pages: " 
            10.1038/s42003-022-04140-y"
COMMENT     Sequence Label: pS238D1::sptse5-CT_phoA-lacZalpha
FEATURES             Location/Qualifiers
     source          1..8573
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(19..984)
                     /codon_start=1
                     /gene="xylS"
                     /product="XylS regulator encoded by the Pseudomonas putida 
                     TOL plasmid pWWO"
                     /label=XylS
                     /note="stimulates transcription from the Pm promoter in the
                     presence of aromatic compounds such as m-toluate or 
                     benzoate"
                     /translation="MDFCLLNEKSQIFVHAEPYAVSDYVNQYVGTHSIRLPKGGRPAGR
                     LHHRIFGCLDLCRISYGGSVRVISPGLETCYHLQIILKGHCLWRGHGQEHYFAPGELLL
                     LNPDDQADLTYSEDCEKFIVKLPSVVLDRACSDNNWHKPREGIRFAARHNLQQLDGFIN
                     LLGLVCDEAEHTKSMPRVQEHYAGIIASKLLEMLGSNVSREIFSKGNPSFERVVQFIEE
                     NLKRNISLERLAELAMMSPRSLYNLFEKHAGTTPKNYIRNRKLESIRACLNDPSANVRS
                     ITEIALDYGFLHLGRFAENYRSAFGELPSDTLRQCKKEVA"
     promoter        1884..1964
                     /label=Pm promoter
                     /note="The bacterial Pm promoter is activated by XylS in 
                     the presence of benzoate or m-toluate (Marques et al., 
                     1999)."
     CDS             2018..3100
                     /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     complement(2037..2056)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     sig_peptide     3135..3200
                     /codon_start=1
                     /gene="pelB (fragment)"
                     /product="leader peptide for secretion"
                     /label=pelB signal sequence
                     /translation="MKYLLPTAAAGLLLLAAQPAMA"
     primer_bind     complement(5027..5044)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5027..5043)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5036..5058)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     terminator      complement(5306..5333)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      complement(5349..5419)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      5605..5699
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(5820..5839)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             5831..6646
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     oriT            6813..6921
                     /note="incP origin of transfer"
     CDS             complement(6932..7594)
                     /codon_start=1
                     /product="replication protein for the broad-host-range 
                     plasmid pBBR1 from Bordetella bronchiseptica"
                     /label=pBBR1 Rep
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
     rep_origin      7595..8366
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     terminator      complement(8481..8567)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ttaattaaac gttcgtaatc aagccacttc ctttttgcat tgacgcaggg tgtcggaagg
       61 caactcgccg aacgcgctcc tatagttttc agcgaagcgt cccaaatgta agaagccgta
      121 gtctagggct atctcagtta tactacgcac attggcactg ggatcgttca agcaggcgcg
      181 gatgctttcg agcttgcggt tgcggatgta gttcttcggc gtggtgccgg cgtgcttctc
      241 gaacaaattg tagagcgagc gtggactcat catcgccagc tccgctaacc gctcaaggct
      301 gatattccgt ttgagattct cctcaatgaa ttgaacgact cgctcgaaag acgggttacc
      361 tttgctgaaa atttcacggc tgacattgct gcccagcatt tcgagcagct tggaagcgat
      421 gatccccgca tagtgctctt ggacccgagg catcgacttt gtatgttccg cttcgtcaca
      481 aactaacccg agtagattga taaagccatc gagttgctgg agattgtgtc gcgcggcgaa
      541 acggataccc tccctcggct tgtgccaatt gttgtcactg cacgcccgat caaggaccac
      601 tgagggcaat ttaacgataa atttctcgca atcttctgaa taggtcaggt cggcttggtc
      661 atccggattg agcagcaata gttcgcccgg cgcaaaatag tgctcctggc catggccacg
      721 ccacaggcaa tggcctttga gtattatttg cagatgataa caggtctcta atccaggcga
      781 gattaccctc acgctaccgc cgtagctgat tcgacacaga tcgaggcatc cgaagattct
      841 gtggtgcagc ctgcctgccg ggcgcccgcc cttgggcagg cgaatagagt gcgtaccgac
      901 atactggtta acataatcgg agactgcata gggctcggcg tggacgaaga tctgactttt
      961 ctcgttcaat aagcaaaaat ccatagttca cggttctctt attttaatgt gggctgcttg
     1021 gtgtgatgta gaaaggcgcc aagtcgatga aaatgcatct cgacgtgatg cgtatacggg
     1081 ttacccccat tgccacgttg cgccatcctt tttgcaatca gtgaccactt ttccaagcaa
     1141 aaataacgcc aagcagaacg aagacgttct ttttaagaag cgagaacacc agaagttcgt
     1201 gctgtcgggg catggggcga cgaattggcg gataaagggg atctgctgga tattacggcc
     1261 tttttaaaga ccgtaaagaa aaataagcac aagttttatc cggcctttat tcacattctt
     1321 gcccgcctga tgaatgctca tccgtaatta cgtatggcaa tgaaagacgg tgagctggtg
     1381 atatgggata gtgttcaccc ttgttacacc gttttccatg agcaaactga aacgttttca
     1441 tcgctctgga gtgaatacca cgacgatttc cggcagtttc tacacatata ttcgcaagat
     1501 gtggcgtgtt acggtgaaaa cctggcctat ttccctaaag ggtttattga gaatatgttt
     1561 ttcgtctcag ccaatccctg ggtgagtttc accagttttg atttaaacgt ggccaatatg
     1621 gacaacttct tcgcccccgt tttcaccatg catgggaatt agcttgatct gaccaacgac
     1681 cggtagcgga gctatccaac ggcggtatac caggaaaaca cacagcaggt acatcagaac
     1741 agtaccatga ctgaagaaca aatagttttt tcctgatcca taaagcagaa cggcctgctc
     1801 catgacaaat ctggctcccc aactaatgcc ccatgcagcc agcataacca gcataaagtg
     1861 cagtgtccgg tttgataggg ataagtccag ccttgcaaga agcggataca ggagtgcaaa
     1921 aaatggctat ctctagtaag gcctacccct taggctttat gcaacagaaa caataataat
     1981 gggagtcatg accatgccta ggaggaaaaa acatatggtg aaaccagtaa cgttatacga
     2041 tgtcgcagag tatgccggtg tctcttatca gaccgtttcc cgcgtggtga accaggccag
     2101 ccacgtttct gcgaaaacgc gggaaaaagt ggaagcggcg atggcggagc tgaattacat
     2161 tcccaaccgc gtggcacaac aactggcggg caaacagtcg ttgctgattg gcgttgccac
     2221 ctccagtctg gccctgcacg cgccgtcgca aattgtcgcg gcgattaaat ctcgcgccga
     2281 tcaactgggt gccagcgtgg tggtgtcgat ggtagaacga agcggcgtcg aagcctgtaa
     2341 agcggcggtg cacaatcttc tcgcgcaacg cgtcagtggg ctgatcatta actatccgct
     2401 ggatgaccag gatgccattg ctgtggaagc tgcctgcact aatgttccgg cgttatttct
     2461 tgatgtctct gaccagacac ccatcaacag tattattttc tcccatgaag acggtacgcg
     2521 actgggcgtg gagcatctgg tcgcattggg tcaccagcaa atcgcgctgt tagcgggccc
     2581 attaagttct gtctcggcgc gtctgcgtct ggctggctgg cataaatatc tcactcgcaa
     2641 tcaaattcag ccgatagcgg aacgggaagg cgactggagt gccatgtccg gttttcaaca
     2701 aaccatgcaa atgctgaatg agggcatcgt tcccactgcg atgctggttg ccaacgatca
     2761 gatggcgctg ggcgcaatgc gcgccattac cgagtccggg ctgcgcgttg gtgcggatat
     2821 ctcggtagtg ggatacgacg ataccgaaga cagctcatgt tatatcccgc cgttaaccac
     2881 catcaaacag gattttcgcc tgctggggca aaccagcgtg gaccgcttgc tgcaactctc
     2941 tcagggccag gcggtgaagg gcaatcagct gttgcccgtc tcactggtga aaagaaaaac
     3001 caccctggcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
     3061 gctggcacga caggtttccc gactggaaag cgggcagtaa catcaccacc atcattagga
     3121 tggtggtgat gataatgaag tacctgctgc cgaccgcggc ggcgggtctg ctgctgctgg
     3181 cggcgcagcc ggcgatggcg atgggcctga tcattccgct ggttgtgatc ggtgcgtttg
     3241 cggcgcgtgc ggcgattggt gcggcgctgg gtgcgggcat tgagctgggt atgcagaccg
     3301 gcaaacaagt gctgggtcag atgaaggata actgggacag cgatcgtgac ctgaccgata
     3361 tcaagtggaa atgcatcgac attaactgga aacatgttgg tgcgagcgcg gcgattggta
     3421 ccgttgcgcc gggcatgctg agcaccggta aaaccgtggt tcagagcgcg aaggcgattc
     3481 gtaccctgag cggtcaggcg gcgaacaccg cgaaccgtgc ggcgaaactg gcggcgcgta
     3541 aggcggcgca cgcggatacc atcaagaaag cggtggcgac ccaagcggcg tggcagaccg
     3601 gcaagcaaat tgttaaatgc ccgctgaagg acgaggaaga ggaatgcccg ccgcaaggga
     3661 gctcggcccg gacaccagaa atgcctgttc tggaaaaccg ggctgctcag ggcgatatta
     3721 ctgcacccgg cggtgctcgc cgtttaacgg gtgatcagac tgccgctctg cgtgattctc
     3781 ttagcgataa acctgcaaaa aatattattt tgctgattgg cgatgggatg ggggactcgg
     3841 aaattactgc cgcacgtaat tatgccgaag gtgcgggcgg cttttttaaa ggtatagatg
     3901 ccttaccgct taccgggcaa tacactcact atgcgctgaa taaaaaaacc ggcaaaccgg
     3961 actacgtcac cgactcggct gcatcagcaa ccgcctggtc aaccggtgtc aaaacctata
     4021 acggcgcgct gggcgtcgat attcacgaaa aagatcaccc aacgattctg gaaatggcaa
     4081 aagccgcagg tctggcgacc ggtaacgttt ctaccgcaga gttgcaggat gccacgcccg
     4141 ctgcgctggt ggcacatgtg acctcgcgca aatgctacgg tccgagcgcg accagtgaaa
     4201 aatgtccggg taacgctctg gaaaaaggcg gaaaaggatc gattaccgaa cagctgctta
     4261 acgctcgtgc cgacgttacg cttggcggcg gcgcaaaaac ctttgctgaa acggcaaccg
     4321 ctggtgaatg gcagggaaaa acgctgcgtg aacaggcaca ggcgcgtggt tatcagttgg
     4381 tgagcgatgc tgcctcactg aattcggtga cggaagcgaa tcagcaaaaa cccctgcttg
     4441 gcctgtttgc tgacggcaat atgccagtgc gctggctagg accgaaagca acgtaccatg
     4501 gcaatatcga taagcccgca gtcacctgta cgccaaatcc gcaacgtaat gacagtgtac
     4561 caaccctggc gcagatgacc gacaaagcca ttgaattgtt gagtaaaaat gagaaaggct
     4621 ttttcctgca agttgaaggt gcgtcaatcg ataaacagga tcatgctgcg aatccttgtg
     4681 ggcaaattgg cgagacggtc gatctcgatg aagccgtaca acgggcgctg gaattcgcta
     4741 aaaaggaggg taacacgctg gtcatagtca ccgctgatca cgcccacgcc agccagattg
     4801 ttgcgccgga taccaaagct ccgggcctca cccaggcgct aaataccaaa gatggcgcag
     4861 tgatggtgat gagttacggg aactccgaag aggattcaca agaacatacc ggcagtcagt
     4921 tgcgtattgc ggcgtatggc ccgcatgccg ccaatgttgt tggactgacc gaccagaccg
     4981 atctcttcta caccatgaaa gccgctctgg ggctgaaaac ggattcactg gccgtcgttt
     5041 tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc
     5101 cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt
     5161 tgcgcagcct gaatggcgaa tggcgctaat gaggtacccg gggatcctct agagtcgacc
     5221 tgcaggcatg caagcttgcg gccgcgtcgt gactgggaaa accctggcgc aaagcccgcc
     5281 gaaaggcggg cttttctgtt ataaacgcag aaaggcccac ccgaaggtga gccagtgtga
     5341 ctctagtaga gagcgttcac cgacaaaaac agataaaacg aaaggcccag tctttcgact
     5401 gagcctttcg ttttatttga tgcctggctc gagctcgaga aaaaaagccc ggacgactgt
     5461 tcgggcttgt ctttttatat gttggaaaat cagtggcaat gcaatggccc aacagaaaat
     5521 ggtgaaacca gtaacgttat actgaatcta agtatcaatg tgagcgctca caagtcaaca
     5581 ctctttttga taaaactagt cttggactcc tgttgataga tccagtaatg acctcagaac
     5641 tccatctgga tttgttcaga acgctcggtt gccgccgggc gttttttatt ggtgagaatc
     5701 caggggtccc caataattac gatttaaatt tgtgtctcaa aatctctgat gttacattgc
     5761 acaagataaa aatatatcat catgaacaat aaaactgtct gcttacataa acagtaatac
     5821 aaggggtgtt atgagccata ttcagcgtga aacgagctgt agccgtccgc gtctgaacag
     5881 caacatggat gcggatctgt atggctataa atgggcgcgt gataacgtgg gtcagagcgg
     5941 cgcgaccatt tatcgtctgt atggcaaacc ggatgcgccg gaactgtttc tgaaacatgg
     6001 caaaggcagc gtggcgaacg atgtgaccga tgaaatggtg cgtctgaact ggctgaccga
     6061 atttatgccg ctgccgacca ttaaacattt tattcgcacc ccggatgatg cgtggctgct
     6121 gaccaccgcg attccgggca aaaccgcgtt tcaggtgctg gaagaatatc cggatagcgg
     6181 cgaaaacatt gtggatgcgc tggccgtgtt tctgcgtcgt ctgcatagca ttccggtgtg
     6241 caactgcccg tttaacagcg atcgtgtgtt tcgtctggcc caggcgcaga gccgtatgaa
     6301 caacggcctg gtggatgcga gcgattttga tgatgaacgt aacggctggc cggtggaaca
     6361 ggtgtggaaa gaaatgcata aactgctgcc gtttagcccg gatagcgtgg tgacccacgg
     6421 cgattttagc ctggataacc tgattttcga tgaaggcaaa ctgattggct gcattgatgt
     6481 gggccgtgtg ggcattgcgg atcgttatca ggatctggcc attctgtgga actgcctggg
     6541 cgaatttagc ccgagcctgc aaaaacgtct gtttcagaaa tatggcattg ataatccgga
     6601 tatgaacaaa ctgcaatttc atctgatgct ggatgaattt ttctaataat taattggacc
     6661 gcggtccgcg cgttgtcctt ttccgctgca taaccctgct tcggggtcat tatagcgatt
     6721 ttttcggtat atccatcctt tttcgcacga tatacaggat tttgccaaag ggttcgtgta
     6781 gactttcctt ggtgtatcca acggcgtcag ccgggcagga taggtgaagt aggcccaccc
     6841 gcgagcgggt gttccttctt cactgtccct tattcgcacc tggcggtgct caacgggaat
     6901 cctgctctgc gaggctggcc gtaggccggc cctaccggcg cggcagcgtt acccgtgtcg
     6961 gcggctccaa cggctcgcca tcgtccagaa aacacggctc atcgggcatc ggcaggcgct
     7021 gctgcccgcg ccgttcccat tcctccgttt cggtcaaggc tggcaggtct ggttccatgc
     7081 ccggaatgcc gggctggctg ggcggctcct cgccggggcc ggtcggtagt tgctgctcgc
     7141 ccggatacag ggtcgggatg cggcgcaggt cgccatgccc caacagcgat tcgtcctggt
     7201 cgtcgtgatc aaccaccacg gcggcactga acaccgacag gcgcaactgg tcgcggggct
     7261 ggccccacgc cacgcggtca ttgaccacgt aggccgacac ggtgccgggg ccgttgagct
     7321 tcacgacgga gatccagcgc tcggccacca agtccttgac tgcgtattgg accgtccgca
     7381 aagaacgtcc gatgagcttg gaaagtgtct tctggctgac caccacggcg ttctggtggc
     7441 ccatctgcgc cacgaggtga tgcagcagca ttgccgccgt gggtttcctc gcaataagcc
     7501 cggcccacgc ctcatgcgct ttgcgttccg tttgcaccca gtgaccgggc ttgttcttgg
     7561 cttgaatgcc gatttctctg gactgcgtgg ccatgcttat ctccatgcgg taggggtgcc
     7621 gcacggttgc ggcaccatgc gcaatcagct gcaacttttc ggcagcgcga caacaattat
     7681 gcgttgcgta aaagtggcag tcaattacag attttcttta acctacgcaa tgagctattg
     7741 cggggggtgc cgcaatgagc tgttgcgtac cccccttttt taagttgttg atttttaagt
     7801 ctttcgcatt tcgccctata tctagttctt tggtgcccaa agaagggcac ccctgcgggg
     7861 ttcccccacg ccttcggcgc ggctccccct ccggcaaaaa gtggcccctc cggggcttgt
     7921 tgatcgactg cgcggccttc ggccttgccc aaggtggcgc tgcccccttg gaacccccgc
     7981 actcgccgcc gtgaggctcg gggggcaggc gggcgggctt cgcccttcga ctgcccccac
     8041 tcgcataggc ttgggtcgtt ccaggcgcgt caaggccaag ccgctgcgcg gtcgctgcgc
     8101 gagccttgac ccgccttcca cttggtgtcc aaccggcaag cgaagcgcgc aggccgcagg
     8161 ccggaggctt ttccccagag aaaattaaaa aaattgatgg ggcaaggccg caggccgcgc
     8221 agttggagcc ggtgggtatg tggtcgaagg ctgggtagcc ggtgggcaat ccctgtggtc
     8281 aagctcgtgg gcaggcgcag cctgtccatc agcttgtcca gcagggttgt ccacgggccg
     8341 agcgaagcga gccagccggt ggccgctcgc ggccatcgtc cacatatcca cgggctggca
     8401 agggagcgca gcgaccgcgc agggcgaagc ccggagagca agcccgtagg gggggcgcgc
     8461 ccagctgtct agggcggcgg atttgtccta ctcaggagag cgttcaccga caaacaacag
     8521 ataaaacgaa aggcccagtc tttcgactga gcctttcgtt ttatttgatg cct
//
