LOCUS       sequence-380604-        4713 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For membrane insertion study; Expresses sptse5-CT (1169-1300) /PhoA 
            (22-472)/LacZ (4-60)fusion with PelB signalpeptide fused at the 
            N-terminus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4713)
  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 4713)
  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: pKTop::sptse5-CT (1169-1300)
FEATURES             Location/Qualifiers
     source          1..4713
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    110..131
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(146..151,152..169,170..176)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    184..200
                     /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     189..211
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     208..224
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     208..224
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     sig_peptide     242..307
                     /codon_start=1
                     /gene="pelB (fragment)"
                     /product="leader peptide for secretion"
                     /label=pelB signal sequence
                     /translation="MKYLLPTAAAGLLLLAAQPAMA"
     primer_bind     complement(2083..2100)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2083..2099)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2092..2114)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2258..2277)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2377..2399
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2717..3511)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2828..2847)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     3438..3457
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     rep_origin      3907..4452
                     /direction=RIGHT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     4353..4372
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4569..4586
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gctagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct
       61 ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt
      121 agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg
      181 gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc
      241 tatgaagtac ctgctgccga ccgcggcggc gggtctgctg ctgctggcgg cgcagccggc
      301 gatggcgatg ggcctgatca ttccgctggt tgtgatcggt gcgtttgcgg cgcgtgcggc
      361 gattggtgcg gcgctgggtg cgggcattga gctgggtatg cagaccggca aacaagtgct
      421 gggtcagatg aaggataact gggacagcga tcgtgacctg accgatatca agtggaaatg
      481 catcgacatt aactggaaac atgttggtgc gagcgcggcg attggtaccg ttgcgccggg
      541 catgctgagc accggtaaaa ccgtggttca gagcgcgaag gcgattcgta ccctgagcgg
      601 tcaggcggcg aacaccgcga accgtgcggc gaaactggcg gcgcgtaagg cggcgcacgc
      661 ggataccatc aagaaagcgg tggcgaccca agcggcgtgg cagaccggca aggggagctc
      721 ggcccggaca ccagaaatgc ctgttctgga aaaccgggct gctcagggcg atattactgc
      781 acccggcggt gctcgccgtt taacgggtga tcagactgcc gctctgcgtg attctcttag
      841 cgataaacct gcaaaaaata ttattttgct gattggcgat gggatggggg actcggaaat
      901 tactgccgca cgtaattatg ccgaaggtgc gggcggcttt tttaaaggta tagatgcctt
      961 accgcttacc gggcaataca ctcactatgc gctgaataaa aaaaccggca aaccggacta
     1021 cgtcaccgac tcggctgcat cagcaaccgc ctggtcaacc ggtgtcaaaa cctataacgg
     1081 cgcgctgggc gtcgatattc acgaaaaaga tcacccaacg attctggaaa tggcaaaagc
     1141 cgcaggtctg gcgaccggta acgtttctac cgcagagttg caggatgcca cgcccgctgc
     1201 gctggtggca catgtgacct cgcgcaaatg ctacggtccg agcgcgacca gtgaaaaatg
     1261 tccgggtaac gctctggaaa aaggcggaaa aggatcgatt accgaacagc tgcttaacgc
     1321 tcgtgccgac gttacgcttg gcggcggcgc aaaaaccttt gctgaaacgg caaccgctgg
     1381 tgaatggcag ggaaaaacgc tgcgtgaaca ggcacaggcg cgtggttatc agttggtgag
     1441 cgatgctgcc tcactgaatt cggtgacgga agcgaatcag caaaaacccc tgcttggcct
     1501 gtttgctgac ggcaatatgc cagtgcgctg gctaggaccg aaagcaacgt accatggcaa
     1561 tatcgataag cccgcagtca cctgtacgcc aaatccgcaa cgtaatgaca gtgtaccaac
     1621 cctggcgcag atgaccgaca aagccattga attgttgagt aaaaatgaga aaggcttttt
     1681 cctgcaagtt gaaggtgcgt caatcgataa acaggatcat gctgcgaatc cttgtgggca
     1741 aattggcgag acggtcgatc tcgatgaagc cgtacaacgg gcgctggaat tcgctaaaaa
     1801 ggagggtaac acgctggtca tagtcaccgc tgatcacgcc cacgccagcc agattgttgc
     1861 gccggatacc aaagctccgg gcctcaccca ggcgctaaat accaaagatg gcgcagtgat
     1921 ggtgatgagt tacgggaact ccgaagagga ttcacaagaa cataccggca gtcagttgcg
     1981 tattgcggcg tatggcccgc atgccgccaa tgttgttgga ctgaccgacc agaccgatct
     2041 cttctacacc atgaaagccg ctctggggct gaaaacggat tcactggccg tcgttttaca
     2101 acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc
     2161 tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg
     2221 cagcctgaat ggcgaatggc gctaagtgca ctctcagtac aatctgctct gatgccgcat
     2281 agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc
     2341 tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt
     2401 tttcaccgtc atcaccgaaa cgcgcgagac gaaagggccg cccttcccaa cagttgcgca
     2461 gcctgaatgg cgaatggcgc tgatgtccgg cggtgctttt gccgttacgc accaccccgt
     2521 cagtagctga acaggaggga cagggggtgg gcgaagaact ccagcatgag atccccgcgc
     2581 tggaggatca tccagccggc gtcccggaaa acgattccga agcccaacct ttcatagaag
     2641 gcggcggtgg aatcgaaatc tcgtgatggc aggttgggcg tcgcttggtc ggtcatttcg
     2701 aaccccagag tcccgctcag aagaactcgt caagaaggcg atagaaggcg atgcgctgcg
     2761 aatcgggagc ggcgataccg taaagcacga ggaagcggtc agcccattcg ccgccaagct
     2821 cttcagcaat atcacgggta gccaacgcta tgtcctgata gcggtccgcc acacccagcc
     2881 ggccacagtc gatgaatcca gaaaagcggc cattttccac catgatattc ggcaagcagg
     2941 catcgccatg ggtcacgacg agatcctcgc cgtcgggcat ccgcgccttg agcctggcga
     3001 acagttcggc tggcgcgagc ccctgatgct cttcgtccag atcatcctga tcgacaagac
     3061 cggcttccat ccgagtacgt gctcgctcga tgcgatgttt cgcttggtgg tcgaatgggc
     3121 aggtagccgg atcaagcgta tgcagccgcc gcattgcatc agccatgatg gatactttct
     3181 cggcaggagc aaggtgagat gacaggagat cctgccccgg cacttcgccc aatagcagcc
     3241 agtcccttcc cgcttcagtg acaacgtcga gcacagctgc gcaaggaacg cccgtcgtgg
     3301 ccagccacga tagccgcgct gcctcgtctt ggagttcatt cagggcaccg gacaggtcgg
     3361 tcttgacaaa aagaaccggg cgcccctgcg ctgacagccg gaacacggcg gcatcagagc
     3421 agccgattgt ctgttgtgcc cagtcatagc cgaatagcct ctccacccaa gcggccggag
     3481 aacctgcgtg caatccatct tgttcaatca tgcgaaacga tcctcatcct gtctcttgat
     3541 cagatcttga tcccctgcgc catcagatcc ttggcggcaa gaaagccatc cagtttactt
     3601 tgcagggctt cccaacctta ccagagggcg ccccagctgg caattccggt tcgcttgctg
     3661 tccataaaac cgcccagtct agctatcgcc atgtaagccc actgcaagct acctgctttc
     3721 tctttgcgct tgcgttttcc cttgtccaga tagcccagta gctgacattc atccggggtc
     3781 agcaccgttt ctgcggactg gctttctacg tgttccgctt cctttagcag cccttgcgcc
     3841 ctgagtgctt gcggcagcgt gaagctgtgc ctagaaatat tttatctgat taataagatg
     3901 atcttcttga gatcgttttg gtctgcgcgt aatctcttgc tctgaaaacg aaaaaaccgc
     3961 cttgcagggc ggtttttcga aggttctctg agctaccaac tctttgaacc gaggtaactg
     4021 gcttggagga gcgcagtcac caaaacttgt cctttcagtt tagccttaac cggcgcatga
     4081 cttcaagact aactcctcta aatcaattac cagtggctgc tgccagtggt gcttttgcat
     4141 gtctttccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcggactgaa
     4201 cggggggttc gtgcatacag tccagcttgg agcgaactgc ctacccggaa ctgagtgtca
     4261 ggcgtggaat gagacaaacg cggccataac agcggaatga caccggtaaa ccgaaaggca
     4321 ggaacaggag agcgcacgag ggagccgcca gggggaaacg cctggtatct ttatagtcct
     4381 gtcgggtttc gccaccactg atttgagcgt cagatttcgt gatgcttgtc aggggggcgg
     4441 agcctatgga aaaacggctt tgccgcggcc ctctcacttc cctgttaagt atcttcctgg
     4501 catcttccag gaaatctccg ccccgttcgt aagccatttc cgctcgccgc agtcgaacga
     4561 ccgagcgtag cgagtcagtg agcgaggaag cggaatatat cctgtatcac atattctgct
     4621 gacgcaccgg tgcagccttt tttctcctgc cacatgaagc acttcactga caccctcatc
     4681 agtgccaaca tagtaagcca gtatatacac tcc
//
