LOCUS       sequence-181677-        4511 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Mariner transposon mutagenesis.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 4511)
  AUTHORS   Bachman MA, Breen P, Deornellas V, Mu Q, Zhao L, Wu W, Cavalcoli JD,
            Mobley HL
  TITLE     Genome-Wide Identification of Klebsiella pneumoniae Fitness Genes 
            during Lung Infection.
  JOURNAL   MBio. 2015 Jun 9;6(3):e00775. doi: 10.1128/mBio.00775-15.
  PUBMED    26060277
REFERENCE   2  (bases 1 to 4511)
  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: "MBio."; 
            date: "2015-06-9"; volume: "6(3):e00775. doi"; pages: " 
            10.1128/mBio.00775-15"
COMMENT     Sequence Label: pSAM_Cam
FEATURES             Location/Qualifiers
     source          1..4511
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     rep_origin      186..573
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     oriT            744..853
                     /note="incP origin of transfer"
     protein_bind    complement(970..1017)
                     /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(1034..1693)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPELRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1608..1627
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(1694..1796)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     protein_bind    complement(1902..1935)
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     CDS             2372..3166
                     /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(2426..2445)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3036..3055
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     terminator      3225..3311
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3403..3430
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     protein_bind    3528..3549
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3564..3569,3570..3587,3588..3594)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
ORIGIN
        1 gcagggaaaa aatttggctg caatgaagag gtgatcgccg aaactgaggc ctattttgag
       61 gcaaaaccga aggagtacta ccaaaatggt atcaaaaaat tggaaggtcg ttataatcgt
      121 tgtatcgctc ttgaagggaa ctatgttgaa taagcggccg ccaccgcggt ggaggggaat
      181 tcccatgtca gccgttaagt gttcctgtgt cactcaaaat tgctttgaga ggctctaagg
      241 gcttctcagt gcgttacatc cctggcttgt tgtccacaac cgttaaacct taaaagcttt
      301 aaaagcctta tatattcttt tttttcttat aaaacttaaa accttagagg ctatttaagt
      361 tgctgattta tattaatttt attgttcaaa catgagagct tagtacgtga aacatgagag
      421 cttagtacgt tagccatgag agcttagtac gttagccatg agggtttagt tcgttaaaca
      481 tgagagctta gtacgttaaa catgagagct tagtacgtga aacatgagag cttagtacgt
      541 actatcaaca ggttgaactg ctgatcttca gatyttccay rcgcgtcaat tcgaggggga
      601 tcaattccgt tttccgctgc ataaccctgc ttcggggtca ttatagcgat tttttcggta
      661 tatccatcct ttttcgcacg atatacagga ttttgccaaa gggttcgtgt agactttcct
      721 tggtgtatcc aacggcgtca gccgggcagg ataggtgaag taggcccacc cgcgagcggg
      781 tgttccttct tcactgtccc ttattcgcac ctggcggtgc tcaacgggaa tcctgctctg
      841 cgaggctggc cggctaccgc cggcgtaaca gatgagggca agcggatggc tgatgaaacc
      901 aagccaacca ggaagggcag cccacctatc acggaattga tccacgcgtg tgtaggctgg
      961 agctgcttcg aagttcctat actttctaga gaataggaac ttcggaatag gaacttcatt
     1021 taaatggcgc gccttacgcc ccgccctgcc actcatcgca gtactgttgt aattcattaa
     1081 gcattctgcc gacatggaag ccatcacaaa cggcatgatg aacctgaatc gccagcggca
     1141 tcagcacctt gtcgccttgc gtataatatt tgcccatggt gaaaacgggg gcgaagaagt
     1201 tgtccatatt ggccacgttt aaatcaaaac tggtgaaact cacccaggga ttggctgaga
     1261 cgaaaaacat attctcaata aaccctttag ggaaataggc caggttttca ccgtaacacg
     1321 ccacatcttg cgaatatatg tgtagaaact gccggaaatc gtcgtggtat tcactccaga
     1381 gcgatgaaaa cgtttcagtt tgctcatgga aaacggtgta acaagggtga acactatccc
     1441 atatcaccag ctcaccgtct ttcattgcca tacgtaattc cggatgagca ttcatcaggc
     1501 gggcaagaat gtgaataaag gccggataaa acttgtgctt atttttcttt acggtcttta
     1561 aaaaggccgt aatatccagc tgaacggtct ggttataggt acattgagca actgactgaa
     1621 atgcctcaaa atgttcttta cgatgccatt gggatatatc aacggtggta tatccagtga
     1681 tttttttctc cattttagct tccttagctc ctgaaaatct cgacaactca aaaaatacgc
     1741 ccggtagtga tcttatttca ttatggtgaa agttggaacc tcttacgtgc cgatcaacgt
     1801 ctcattttcg ccaaaagttg gcccagggct tcccggtatc aacagggaca ccaggattta
     1861 tttattctgc gaagtgatct tccgtcacag gtaggcgcgc cgaagttcct atactttcta
     1921 gagaatagga acttcggaat aggaactaag gaggatattc acatggacca tggctaattc
     1981 ccatacgcgt caattcgagg gggatcaatt ccgttttccg ctgcataacc ctgcttcggg
     2041 gtcatcatts sagcwatttm ggwagmksaa cccgtagaaa gccagtccgc agaaacggtg
     2101 ctgaccccgg atgaatgtca gctactgggc tatctggaca agggaaaacg caagcgcaaa
     2161 gagaaagcag gtagcttgca gtgggcttac atggcgatag ctagactggg cggttttatg
     2221 gacagcaagc gaaccggaat tgccagctgg ggcgccctct ggtaaggttg ggaagccctg
     2281 caaagtaaac tggatggctt tcttgccgcc aaggatctga tggcgcaggg gatcaagatc
     2341 tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg
     2401 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg
     2461 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa
     2521 gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct
     2581 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga
     2641 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc
     2701 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac
     2761 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc
     2821 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
     2881 gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga
     2941 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
     3001 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga
     3061 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
     3121 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg
     3181 ttcgaaatga ctctagaatg cgagagtagg gaactgccag gcatcaaata aaacgaaagg
     3241 ctcagtcgaa agactgggcc tttcgtttta tctgttgttt gtcggtgaac gctctcctga
     3301 gtaggacaaa tccgccggga gcggatttga acgttgcgaa gcaacggccc ggagggtggc
     3361 gggcaggacg cccgccataa actgccaggc atcaaattaa gcagaaggcc atcctgacgg
     3421 atggcctttt tgcgtttcta cctgcagggc gcgccaagca gaagacggca tacgaagacc
     3481 ggggacttat catccaacct gttaggatcc agtgagcgca acgcaattaa tgtgagttag
     3541 ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga
     3601 attgtgagcg gatacatatg gaaaaaaagg aatttcgtgt tttgataaaa tactgttttc
     3661 tgaagggaaa aaatacagtg gaagcaaaaa cttggcttga taatgagttt ccggactctg
     3721 ccccagggaa atcaacaata attgattggt atgcaaaatt caagcgtggt gaaatgagca
     3781 cggaggacgg tgaacgcagt ggacgcccga aagaggtggt taccgacgaa aacatcaaaa
     3841 aaatccacaa aatgattttg aatgaccgta aaatgaagtt gatcgagata gcagaggcct
     3901 taaagatatc aaaggaacgt gttggtcata tcattcatca atatttggat atgcggaagc
     3961 tctgtgcgaa atgggtgccg cgcgagctca catttgacca aaaacaacga cgtgttgatg
     4021 attctaagcg gtgtttgcag ctgttaactc gtaatacacc cgagtttttc cgtcgatatg
     4081 tgacaatgga tgaaacatgg ctccatcact acactcctga gtccaatcga cagtcggctg
     4141 agtggacagc gaccggtgaa ccgtctccga agcgtggaaa gactcaaaag tccgctggca
     4201 aagtaatggc ctctgttttt tgggatgcgc atggaataat ttttatcgat tatcttgaga
     4261 agggaaaaac catcaacagt gactattata tggcgttatt ggagcgtttg aaggtcgaaa
     4321 tcgcggcaaa acggccccac atgaagaaga aaaaagtgtt gttccaccaa gacaacgcac
     4381 cgtgccacaa gtcattgaga acgatggcaa aaattcatga attgggcttc gaattgcttc
     4441 cccacccgcc gtattctcca gatctggccc ccagcgactt tttcttgttc tcagacctca
     4501 aaaggatgct c
//
