LOCUS       sequence-124260-        8859 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Suicide plasmid for efficient gene mutation in Gram-positive 
            bacteria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8859)
  AUTHORS   Abdelhamed H, Lawrence ML, Karsi A
  TITLE     A novel suicide plasmid for efficient gene mutation in Listeria 
            monocytogenes.
  JOURNAL   Plasmid. 2015 May 31;81:1-8. doi: 10.1016/j.plasmid.2015.05.003.
  PUBMED    26038185
REFERENCE   2  (bases 1 to 8859)
  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: "Plasmid."; 
            date: "2015-05-31"; volume: "81:1-8. doi"; pages: " 
            10.1016/j.plasmid.2015.05.003"
COMMENT     Sequence Label: pHoss1
FEATURES             Location/Qualifiers
     source          1..8859
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(4069..4695)
                     /codon_start=1
                     /gene="tetR"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MMSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWH
                     VKNKRALLDALAIEMLDRRHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGT
                     RPTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPT
                     TDSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     4600..4619
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     protein_bind    4717..4735
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O1 for the tetR and tetA genes"
     protein_bind    4802..4820
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O1 for the tetR and tetA genes"
     CDS             6416..6607
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(6601..6623)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    6709..6849
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     6764..6783
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6864..6881)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7035..7623)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7115..7134)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7794..8585,8586..8654))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     8417..8436
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8655..8759)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8827..8845
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 gaattaattc tcatgtttga cagcttatca tcgattcaca aaaaataggc acacgaaaaa
       61 caagttaagg gatgcagttt atgcatccct taacttactt attaaataat ttatagctat
      121 tgaaaagaga taagaattgt tcaaagctaa tattgtttaa atcgtcaatt cctgcatgtt
      181 ttaaggaatt gttaaattga ttttttgtaa atattttctt gtattctttg ttaacccatt
      241 tcataacgaa ataattatac ttttgtttat ctttgtgtga tattcttgat ttttttctac
      301 ttaatctgat aagtgagcta ttcactttag gtttaggatg aaaatattct cttggaacca
      361 tacttaatat agaaatatca acttctgcca ttaaaagtaa tgccaatgag cgttttgtat
      421 ttaataatct tttagcaaac ccgtattcca cgattaaata aatctcatta gctatactat
      481 caaaaacaat tttgcgtatt atatccgtac ttatgttata aggtatatta ccatatattt
      541 tataggattg gtttttagga aatttaaact gcaatatatc cttgtttaaa acttggaaat
      601 tatcgtgatc aacaagttta ttttctgtag ttttgcataa tttatggtct atttcaatgg
      661 cagttacgaa attacacctc tttactaatt caagggtaaa atggcctttt cctgagccga
      721 tttcaaagat attatcatgt tcatttaatc ttatatttgt cattatttta tctatattat
      781 gttttgaagt aataaagttt tgactgtgtt ttatattttt ctcgttcatt ataaccctct
      841 ttaatttggt tatatgaatt ttgcttatta acgattcatt ataaccactt attttttgtt
      901 tggttgataa tgaactgtgc tgattacaaa aatactaaaa atgcccatat tttttcctcc
      961 ttataaaatt agtataatta tagcacgagc tctgataaat atgaacatga tgagtgatcg
     1021 ttaaatttat actgcaatcg gatgcgatta ttgaataaaa gatatgagag atttatctaa
     1081 tttctttttt cttgtaaaaa aagaaagttc ttaaaggttt tatagttttg gtcgtagagc
     1141 acacggttta acgacttaat tacgaagtaa ataagtctag tgtgttagac tttatgaaat
     1201 ctatatacgt ttatatatat ttattatccg gaggtgtagc atgtctcatt caattttgag
     1261 ggttgccaga gttaaaggat caagtaatac aaacgggata caaagacata atcaaagaga
     1321 gaataaaaac tataataata aagacataaa tcatgaggaa acatataaaa attatgattt
     1381 gattaacgca caaaatataa agtataaaga taaaattgat gaaacgattg atgagaatta
     1441 ttcagggaaa cgtaaaattc ggtcagatgc aattcgacat gtggacggac tggttacaag
     1501 tgataaagat ttctttgatg atttaagcgg agaagaaata gaacgatttt ttaaagatag
     1561 cttggagttt ctagaaaatg aatacggtaa ggaaaatatg ctgtatgcga ctgtccatct
     1621 ggatgaaaga gtcccacata tgcactttgg ttttgtccct ttaacagagg acgggagatt
     1681 gtctgcaaaa gaacagttag gcaacaagaa agactttact caattacaag atagatttaa
     1741 tgagtatgtg aatgagaaag gttatgaact tgaaagaggc acgtccaaag aggttacaga
     1801 acgagaacat aaagcgatgg atcagtacaa gaaagatact gtatttcata aacaggaact
     1861 gcaagaagtt aaggatgagt tacagaaggc aaataagcag ttacagagtg gaatagagca
     1921 tatgaggtct acgaaaccct ttgattatga aaatgagcgt acaggtttgt tctctggacg
     1981 tgaagagact ggtagaaaga tattaactgc tgatgaattt gaacgcctgc aagaaacaat
     2041 ctcttctgca gaacggattg ttgatgatta cgaaaatatt aagagcacag actattacac
     2101 agaaaatcaa gaattaaaaa aacgtagaga gagtttgaaa gaagtagtga atacatggaa
     2161 agaggggtat cacgaaaaaa gtaaagaggt taataaatta aagcgagaga atgatagttt
     2221 gaatgagcag ttgaatgtat cagagaaatt tcaagctagt acagtgactt tatatcgtgc
     2281 tgcgagggcg aatttccctg ggtttgagaa agggtttaat aggcttaaag agaaattctt
     2341 taatgattcc aaatttgagc gtgtgggaca gtttatggat gttgtacagg ataatgtcca
     2401 gaaggtcgat agaaagcgtg agaaacagcg tacagacgat ttagagatgt agaggtactt
     2461 ttatgccgag aaaacttttt gcgtgtgaca gtccttaaaa tatacttaga gcgtaagcga
     2521 aagtagtagc gacagctatt aactttcggt tgcaaagctc taggattttt aatggacgca
     2581 gcgcatcaca cgcaaaaagg aaattggaat aaatgcgaaa tttgagatgt taattaaaga
     2641 cctttttgag gtcttttttt cttagatttt tggggttatt taggggagaa aacatagggg
     2701 ggtactacga cctcccccct aggtgtccat tgtccattgt ccaaacaaat aaataaatat
     2761 tgggttttta atgttaaaag gttgtttttt atgttaaagt gaaaaaaaca gatgttggga
     2821 ggtacagtga tagttgtaga tagaaaagaa gagaaaaaag ttgctgttac tttaagactt
     2881 acaacagaag aaaatgagat attaaataga atcaaagaaa aatataatat tagcaaatca
     2941 gatgcaaccg gtattctaat aaaaaaatat gcaaaggagg aatacggtgc attttaaaca
     3001 aaaaaagata gacagcactg gcatgctgcc tatctatgac taaattttgt taagtgtatt
     3061 agcaccgtta ttatatcatg agcgaaaatg taataaaaga aactgaaaac aagaaaaatt
     3121 caagaggacg taattggaca tttgttttat atccagaatc agcaaaagcc gagtggttag
     3181 agtatttaaa agagttacac attcaatttg tagtgtctcc attacatgat agggatactg
     3241 atacagaagg taggatgaaa aaagagcatt atcatattct agtgatgtat gagggtaata
     3301 aatcttatga acagataaaa ataattacag aagaattgaa tgcgactatt ccgcagattg
     3361 caggaagtgt gaaaggtctt gtgagatata tgcttcacat ggacgatcct aataaattta
     3421 aatatcaaaa agaagatatg atagtttatg gcggtgtaga tgttgatgaa ttattaaaga
     3481 aaacaacaac agatagatat aaattaatta aagaaatgat tgagtttatt gatgaacaag
     3541 gaatcgtaga atttaagagt ttaatggatt atgcaatgaa gtttaaattt gatgattggt
     3601 tcccgctttt atgtgataac tcggcgtatg ttattcaaga atatataaaa tcaaatcggt
     3661 ataaatctga ccgatagatt ttgaatttaa gagtgtcaca agacactctt ttttcgcacc
     3721 aacgaaaact ggtttaagcc gactgcgcaa aagacataat cgataagctt taatgcggta
     3781 gtttatcaca gttaaattgc taacgcagtc aggcaccgtg tatgaaatct aacaatgcgc
     3841 tcatcgtcat cctcggcacc gtcaccctgg atgctgtagg cataggcttg gttatgccgg
     3901 tactgccggg cctcttgcgg gatcgaggcc tttagcatat tatgttgcca actgtcggaa
     3961 cgagacttct ctaaaccttc ccggcttcat catgctctct taacactgct acatcaaccc
     4021 cttgaatcga tcttgaagtt accatcacgg aaaaaggtta tgctgctttt aagacccact
     4081 ttcacattta agttgttttt ctaatccgca tatgatcaat tcaaggccga ataagaaggc
     4141 tggctctgca ccttggtgat caaataattc gatagcttgt cgtaataatg gcggcatact
     4201 atcagtagta ggtgtttccc tttcttcttt agcgacttga tgctcttgat cttccaatac
     4261 gcaacctaaa gtaaaatgcc ccacagcgct gagtgcatat aatgcattct ctagtgaaaa
     4321 accttgttgg cataaaaagg ctaattgatt ttcgagagtt tcatactgtt tttctgtagg
     4381 ccgtgtacct aaatgtactt ttgctccatc gcgatgactt agtaaagcac atctaaaact
     4441 tttagcgtta ttacgtaaaa aatcttgcca gctttcccct tctaaagggc aaaagtgagt
     4501 atggcgccta tctaacatct caatggctaa ggcgtcgagc aaagcccgct tattttttac
     4561 atgccaatac aatgtaggct gctctacacc tagcttctgg gcgagtttac gggttgttaa
     4621 accttcgatt ccgacctcat taagcagctc taatgcgctg ttaatcactt tacttttatc
     4681 taatctagac atcattaatt cctccttttt gttgacactc tatcattgat agagttattt
     4741 gtcaaactag ttttttattt ggatcccctc gagttcatga aaaactaaaa aaaatattga
     4801 cactctatca ttgatagagt ataattaaaa taagcttgat gggaatatga taatagaaat
     4861 accattacca acaccgaact gagtgatttg atcaccaagc catattaaga aagcagttcc
     4921 tgctgtcaaa actagtgcta ttaataaata actcataatt gactgattga taatcagcgc
     4981 acctttgaga taattattaa attggaatgc catacctata gattggataa atgctaaaga
     5041 aattgctaaa taacgagtaa cgttatttaa ctttcttcta cctacttcac cttgttttgc
     5101 ccattctgag aatttaggga caatatccat ttgtaataat tgcattacga ttgatgcagt
     5161 gatgtagggt acaataccca ttgcaaaaat agaaaatcgt ttcaaggctc cgccaccaaa
     5221 agtatttaat aactcagtgg caccttgaga accttgggga ttatcaaaag ctgcaggatt
     5281 tactcctgga gctggtatat aagtccctat tttaaaaatt actaacattg ctagtgtgaa
     5341 gaaaatcttg ttacgaacct cttttgttct aaagaagttc acaagggttt gaatcattag
     5401 atcccatggt acccgggagc tcgaattcta gaagcttctg cagacgcgtc gacgtcatat
     5461 ggatccgata tcgcccgacg cgaggctgga tggccttccc cattatgatt cttctcgctt
     5521 ccggcggcat cgggatgccc gcgttgcagg ccatgctgtc caggcaggta gatgacgacc
     5581 atcagggaca gcttcaagga tcgctcgcgg ctcttaccag cctaacttcg atcattggac
     5641 cgctgatcgt cacggcgatt tatgccgcct cggcgagcac atggaacggg ttggcatgga
     5701 ttgtaggcgc cgccctatac cttgtctgcc tccccgcgtt gcgtcgcggt gcatggagcc
     5761 gggccacctc gacctgaatg gaagccggcg gcacctcgct aacggattca ccactccaag
     5821 aattggagcc aatcaattct tgcggagaac tgtgaatgcg caaaccaacc cttggcagaa
     5881 catatccatc gcgtccgcca tctccagcag ccgcacgcgg cgcatctcgg gcagcgttgg
     5941 gtcctggcca cgggtgcgca tgatcgtgct cctgtcgttg aggacccggc taggctggcg
     6001 gggttgcctt actggttagc agaatgaatc accgatacgc gagcgaacgt gaagcgactg
     6061 ctgctgcaaa acgtctgcga cctgagcaac aacatgaatg gtcttcggtt tccgtgtttc
     6121 gtaaagtctg gaaacgcgga agtcagcgcc ctgcaccatt atgttccgga tctgcatcgc
     6181 aggatgctgc tggctaccct gtggaacacc tacatctgta ttaacgaagc gctggcattg
     6241 accctgagtg atttttctct ggtcccgccg catccatacc gccagttgtt taccctcaca
     6301 acgttccagt aaccgggcat gttcatcatc agtaacccgt atcgtgagca tcctctctcg
     6361 tttcatcggt atcattaccc ccatgaacag aaatccccct tacacggagg catcagtgac
     6421 caaacaggaa aaaaccgccc ttaacatggc ccgctttatc agaagccaga cattaacgct
     6481 tctggagaaa ctcaacgagc tggacgcgga tgaacaggca gacatctgtg aatcgcttca
     6541 cgaccacgct gatgagcttt accgcagctg cctcgcgcgt ttcggtgatg acggtgaaaa
     6601 cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag
     6661 cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggcg cagccatgac
     6721 ccagtcacgt agcgatagcg gagtgtatac tggcttaact atgcggcatc agagcagatt
     6781 gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac
     6841 cgcatcaggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     6901 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     6961 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     7021 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     7081 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     7141 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     7201 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     7261 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     7321 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     7381 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     7441 ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg
     7501 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     7561 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     7621 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     7681 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     7741 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     7801 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     7861 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     7921 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     7981 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     8041 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     8101 gccattgctg caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     8161 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     8221 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     8281 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     8341 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     8401 ccggcgtcaa cacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     8461 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     8521 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     8581 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     8641 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     8701 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     8761 acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc
     8821 tataaaaata ggcgtatcac gaggcccttt cgtcttcaa
//
