LOCUS       sequence-119855-        5272 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  reverse IMCE donor plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 5272)
  TITLE     IMCE attB donor plasmid
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5272)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pJH143
FEATURES             Location/Qualifiers
     source          1..5272
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        69..514
                     /gene="sacR"
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     CDS             join(515..601,602..1936)
                     /codon_start=1
                     /gene="Bacillus subtilis sacB"
                     /product="secreted levansucrase that renders bacterial 
                     growth sensitive to sucrose"
                     /label=SacB
                     /note="negative selection marker"
                     /translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGISH
                     ITRHDMLQIPEQQKNEKYQVSEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYH
                     IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW
                     SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG
                     KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA
                     YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE
                     IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK
                     MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV
                     VKDSILEQGQLTVNK"
     CDS             2594..3385
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     CDS             3410..5218
                     /codon_start=1
                     /gene="uidA (or gusA) from E. coli"
                     /product="beta-glucuronidase"
                     /label=GUS
                     /translation="MLRPVETPTREIKKLDGLWAFSLDRENCGIDQRWWESALQESRAI
                     AVPGSFNDQFADADIRNYAGNVWYQREVFIPKGWAGQRIVLRFDAVTHYGKVWVNNQEV
                     MEHQGGYTPFEADVTPYVIAGKSVRITVCVNNELNWQTIPPGMVITDENGKKKQSYFHD
                     FFNYAGIHRSVMLYTTPNTWVDDITVVTHVAQDCNHASVDWQVVANGDVSVELRDADQQ
                     VVATGQGTSGTLQVVNPHLWQPGEGYLYELCVTAKSQTECDIYPLRVGIRSVAVKGEQF
                     LINHKPFYFTGFGRHEDADLRGKGFDNVLMVHDHALMDWIGANSYRTSHYPYAEEMLDW
                     ADEHGIVVIDETAAVGFNLSLGIGFEAGNKPKELYSEEAVNGETQQAHLQAIKELIARD
                     KNHPSVVMWSIANEPDTRPQVHGNISPLAEATRKLDPTRPITCVNVMFCDAHTDTISDL
                     FDVLCLNRYYGWYVQSGDLETAEKVLEKELLAWQEKLHQPIIITEYGVDTLAGLHSMYT
                     DMWSEEYQCAWLDMYHRVFDRVSAVVGEQVWNFADFATSQGILRVGGNKKGIFTRDRKP
                     KSAAFLLQKRWTGMNFGEKPQQGGKQ"
ORIGIN
        1 cataagcttg tgccagggcg tgcccttgag ttctctcagt tgggggggat cgatcctttt
       61 taacccatca catatacctg ccgttcacta ttatttagtg aaatgagata ttatgatatt
      121 ttctgaattg tgattaaaaa ggcaacttta tgcccatgca acagaaacta taaaaaatac
      181 agagaatgaa aagaaacaga tagatttttt agttctttag gcccgtagtc tgcaaatcct
      241 tttatgattt tctatcaaac aaaagaggaa aatagaccag ttgcaatcca aacgagagtc
      301 taatagaatg aggtcgaaaa gtaaatcgcg cgggtttgtt actgataaag caggcaagac
      361 ctaaaatgtg taaagggcaa agtgtatact ttggcgtcac cccttacata ttttaggtct
      421 ttttttattg tgcgtaacta acttgccatc ttcaaacagg agggctggaa gaagcagacc
      481 gctaacacag tacataaaaa aggagacatg aacgatgaac atcaaaaagt ttgcaaaaca
      541 agcaacagta ttaaccttta ctaccgcact gctggcagga ggcgcaactc aagcgtttgc
      601 gaaagaaacg aaccaaaagc catataagga aacatacggc atttcccata ttacacgcca
      661 tgatatgctg caaatccctg aacagcaaaa aaatgaaaaa tatcaagttt ctgaatttga
      721 ttcgtccaca attaaaaata tctcttctgc aaaaggcctg gacgtttggg acagctggcc
      781 attacaaaac gctgacggca ctgtcgcaaa ctatcacggc taccacatcg tctttgcatt
      841 agccggagat cctaaaaatg cggatgacac atcgatttac atgttctatc aaaaagtcgg
      901 cgaaacttct attgacagct ggaaaaacgc tggccgcgtc tttaaagaca gcgacaaatt
      961 cgatgcaaat gattctatcc taaaagacca aacacaagaa tggtcaggtt cagccacatt
     1021 tacatctgac ggaaaaatcc gtttattcta cactgatttc tccggtaaac attacggcaa
     1081 acaaacactg acaactgcac aagttaacgt atcagcatca gacagctctt tgaacatcaa
     1141 cggtgtagag gattataaat caatctttga cggtgacgga aaaacgtatc aaaatgtaca
     1201 gcagttcatc gatgaaggca actacagctc aggcgacaac catacgctga gagatcctca
     1261 ctacgtagaa gataaaggcc acaaatactt agtatttgaa gcaaacactg gaactgaaga
     1321 tggctaccaa ggcgaagaat ctttatttaa caaagcatac tatggcaaaa gcacatcatt
     1381 cttccgtcaa gaaagtcaaa aacttctgca aagcgataaa aaacgcacgg ctgagttagc
     1441 aaacggcgct ctcggtatga ttgagctaaa cgatgattac acactgaaaa aagtgatgaa
     1501 accgctgatt gcatctaaca cagtaacaga tgaaattgaa cgcgcgaacg tctttaaaat
     1561 gaacggcaaa tggtacctgt tcactgactc ccgcggatca aaaatgacga ttgacggcat
     1621 tacgtctaac gatatttaca tgcttggtta tgtttctaat tctttaactg gcccatacaa
     1681 gccgctgaac aaaactggcc ttgtgttaaa aatggatctt gatcctaacg atgtaacctt
     1741 tacttactca cacttcgctg tacctcaagc gaaaggaaac aatgtcgtga ttacaagcta
     1801 tatgacaaac agaggattct acgcagacaa acaatcaacg tttgcgccga gcttcctgct
     1861 gaacatcaaa ggcaagaaaa catctgttgt caaagacagc atccttgaac aaggacaatt
     1921 aacagttaac aaataaaaac gcaaaagaaa atgccgatgg gtaccgagcg aaatgaccga
     1981 ccaagcgacg cccaacgttt catgttaaac catgcccgtt ccatacagaa gctgggcgaa
     2041 caaacgatgc tcgccttcca gaaaaccgag gatgcgaacc acttcatccg gggtcagcac
     2101 caccggcaag cgccgcgacg gccgaggtct tccgatctcc tgaagccagg gcagatccgt
     2161 gcacagcacc ttgccgtaga agaacagcaa ggccgccaat gcctgacgat gcgtggagac
     2221 cgaaaccttg cgctcgttcg ccagccagga cagaaatgcc tcgacttcgc tgctgcccaa
     2281 ggttgccggg tgacgcacac cgtggaaacg gatgaaggca cgaacccagt ggacataagc
     2341 ctgttcggtt cgtaagctgt aatgcaagta gcgtatgcgc tcacgcaact ggtccagaac
     2401 cttgaccgaa cgcagcggtg gtaacggcgc agtggcggtt ttcatggctt gttatgactg
     2461 tttttttggg gtacagtcta tgcctcgggc atccaagcag caagcgcgtt acgccgtggg
     2521 tcgatgtttg atgttatgga gcagcaacga tgttacgcag cagggcagtc gccctaaaac
     2581 aaagttaaac atcatgaggg aagcggtgat cgccgaagta tcgactcaac tatcagaggt
     2641 agttggcgtc atcgagcgcc atctcgaacc gacgttgctg gccgtacatt tgtacggctc
     2701 cgcagtggat ggcggcctga agccacacag tgatattgat ttgctggtta cggtgaccgt
     2761 aaggcttgat gaaacaacgc ggcgagcttt gatcaacgac cttttggaaa cttcggcttc
     2821 ccctggagag agcgagattc tccgcgctgt agaagtcacc attgttgtgc acgacgacat
     2881 cattccgtgg cgttatccag ctaagcgcga actgcaattt ggagaatggc agcgcaatga
     2941 cattcttgca ggtatcttcg agccagccac gatcgacatt gatctggcta tcttgctgac
     3001 aaaagcaaga gaacatagcg ttgccttggt aggtccagcg gcggaggaac tctttgatcc
     3061 ggttcctgaa caggatctat ttgaggcgct aaatgaaacc ttaacgctat ggaactcgcc
     3121 gcccgactgg gctggcgatg agcgaaatgt agtgcttacg ttgtcccgca tttggtacag
     3181 cgcagtaacc ggcaaaatcg cgccgaagga tgtcgctgcc gactgggcaa tggagcgcct
     3241 gccggcccag tatcagcccg tcatacttga agctagacag gcttatcttg gacaagaaga
     3301 agatcgcttg gcctcgcgcg cagatcagtt ggaagaattt gtccactacg tgaaaggcga
     3361 gatcaccaag gtagtcggca aataatctag agtcgacgag gagtccctta tgttacgtcc
     3421 tgtagaaacc ccaacccgtg aaatcaaaaa actcgacggc ctgtgggcat tcagtctgga
     3481 tcgcgaaaac tgtggaattg atcagcgttg gtgggaaagc gcgttacaag aaagccgggc
     3541 aattgctgtg ccaggcagtt ttaacgatca gttcgccgat gcagatattc gtaattatgc
     3601 gggcaacgtc tggtatcagc gcgaagtctt tataccgaaa ggttgggcag gccagcgtat
     3661 cgtgctgcgt ttcgatgcgg tcactcatta cggcaaagtg tgggtcaata atcaggaagt
     3721 gatggagcat cagggcggct atacgccatt tgaagccgat gtcacgccgt atgttattgc
     3781 cgggaaaagt gtacgtatca ccgtttgtgt gaacaacgaa ctgaactggc agactatccc
     3841 gccgggaatg gtgattaccg acgaaaacgg caagaaaaag cagtcttact tccatgattt
     3901 ctttaactat gccgggatcc atcgcagcgt aatgctctac accacgccga acacctgggt
     3961 ggacgatatc accgtggtga cgcatgtcgc gcaagactgt aaccacgcgt ctgttgactg
     4021 gcaggtggtg gccaatggtg atgtcagcgt tgaactgcgt gatgcggatc aacaggtggt
     4081 tgcaactgga caaggcacta gcgggacttt gcaagtggtg aatccgcacc tctggcaacc
     4141 gggtgaaggt tatctctatg aactgtgcgt cacagccaaa agccagacag agtgtgatat
     4201 ctacccgctt cgcgtcggca tccggtcagt ggcagtgaag ggcgaacagt tcctgattaa
     4261 ccacaaaccg ttctacttta ctggctttgg tcgtcatgaa gatgcggact tacgtggcaa
     4321 aggattcgat aacgtgctga tggtgcacga ccacgcatta atggactgga ttggggccaa
     4381 ctcctaccgt acctcgcatt acccttacgc tgaagagatg ctcgactggg cagatgaaca
     4441 tggcatcgtg gtgattgatg aaactgctgc tgtcggcttt aacctctctt taggcattgg
     4501 tttcgaagcg ggcaacaagc cgaaagaact gtacagcgaa gaggcagtca acggggaaac
     4561 tcagcaagcg cacttacagg cgattaaaga gctgatagcg cgtgacaaaa accacccaag
     4621 cgtggtgatg tggagtattg ccaacgaacc ggatacccgt ccgcaagtgc acgggaatat
     4681 ttcgccactg gcggaagcaa cgcgtaaact cgacccgacg cgtccgatca cctgcgtcaa
     4741 tgtaatgttc tgcgacgctc acaccgatac catcagcgat ctctttgatg tgctgtgcct
     4801 gaaccgttat tacggatggt atgtccaaag cggcgatttg gaaacggcag agaaggtact
     4861 ggaaaaagaa cttctggcct ggcaggagaa actgcatcag ccgattatca tcaccgaata
     4921 cggcgtggat acgttagccg ggctgcactc aatgtacacc gacatgtgga gtgaagagta
     4981 tcagtgtgca tggctggata tgtatcaccg cgtctttgat cgcgtcagcg ccgtcgtcgg
     5041 tgaacaggta tggaatttcg ccgattttgc gacctcgcaa ggcatattgc gcgttggcgg
     5101 taacaagaaa gggatcttca ctcgcgaccg caaaccgaag tcggcggctt ttctgctgca
     5161 aaaacgctgg actggcatga acttcggtga aaaaccgcag cagggaggca aacaatgagt
     5221 ttaaaccccc caactgagag aactcaaggg cacgccctgg cacgaattca tg
//
