LOCUS       sequence-226896-        4099 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Erythromycin resistance cassette.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4099)
  AUTHORS   Diebold-Durand ML, Burmann F, Gruber S
  TITLE     High-Throughput Allelic Replacement Screening in Bacillus subtilis.
  JOURNAL   Methods Mol Biol. 2019;2004:49-61. doi: 10.1007/978-1-4939-9520-2_5.
  PUBMED    31147909
REFERENCE   2  (bases 1 to 4099)
  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: "Methods Mol
            Biol."; date: "2019"; volume: "2004:49-61. doi"; pages: " 
            10.1007/978-1-4939-9520-2_5"
COMMENT     Sequence Label: pJet1.2 ermB cassette
FEATURES             Location/Qualifiers
     source          1..4099
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     305..324
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        305..323
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(join(1931..1937,1938..1955,1956..1961))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    1976..1997
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2117..2134)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2288..2876)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2368..2387)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3047..3838,3839..3907))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     3670..3689
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3908..4012)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4080..4098
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 gcccctgcag ccgaattata ttatttttgc caaataattt ttaacaaaag ctctgaagtc
       61 ttcttcattt aaattcttag atgatacttc atctggaaaa ttgtcccaat tagtagcatc
      121 acgctgtgag taagttctaa accatttttt tattgttgta ttatctctaa tcttactact
      181 cgatgagttt tcggtattat ctctattttt aacttggagc aggttccatt cattgttttt
      241 ttcatcatag tgaataaaat caactgcttt aacacttgtg cctgaacacc atatccatcc
      301 ggcgtaatac gactcactat agggagagcg gccgccagat cttccggatg gctcgagttt
      361 ttcagcaaga tggtctcgaa tatagaagca aacttaagag tgtgttgata gtgcattatc
      421 ttaaaatttt gtataatagg aattgaagtt aaattagatg ctaaaaattt gtaattaaga
      481 aggagggatt cgtcatgttg gtattccaaa tgcgtaatgt agataaaaca tctactgttt
      541 tgaaacagac taaaaacagt gattacgcag ataaataaat acgttagatt aattcctacc
      601 agtgactaat cttatgactt tttaaacaga taactaaaat tacaaacaaa tcgtttaact
      661 tctgtattta tttatagatg taatcacttc aggagtgatt acatgaacaa aaatataaaa
      721 tattctcaaa actttttaac gagtgaaaaa gtactcaacc aaataataaa acaattgaat
      781 ttaaaagaaa ccgataccgt ttacgaaatt ggaacaggta aagggcattt aacgacgaaa
      841 ctggctaaaa taagtaaaca ggtaacgtct attgaattag acagtcatct attcaactta
      901 tcgtcagaaa aattaaaact gaacattcgt gtcactttaa ttcaccaaga tattctacag
      961 tttcaattcc ctaacaaaca gaggtataaa attgttggga atattcctta ccatttaagc
     1021 acacaaatta ttaaaaaagt ggtttttgaa agccatgcgt ctgacatcta tctgattgtt
     1081 gaagaaggat tctacaagcg taccttggat attcaccgaa cactagggtt gctcttgcac
     1141 actcaagtct cgattcagca attgcttaag ctgccagcgg aatgctttca tcctaaacca
     1201 aaagtaaaca gtgtcttaat aaaacttacc cgccatacca cagatgttcc agataaatat
     1261 tggaagctat atacgtactt tgtttcaaaa tgggtcaatc gagaatatcg tcaactgttt
     1321 actaaaaatc agtttcatca agcaatgaaa cacgccaaag taaacaattt aagtaccgtt
     1381 acttatgagc aagtattgtc tatttttaat agttatctat tatttaacgg gaggaaataa
     1441 gaaaaaggcc ccaacatctt ggggcctttt tcttttttat ctacgagacc caatgaatct
     1501 ttctagaaga tctcctacaa tattctcagc tgccatggaa aatcgatgtt cttcttttat
     1561 tctctcaaga ttttcaggct gtatattaaa acttatatta agaactatgc taaccacctc
     1621 atcaggaacc gttgtaggtg gcgtgggttt tcttggcaat cgactctcat gaaaactacg
     1681 agctaaatat tcaatatgtt cctcttgacc aactttattc tgcatttttt ttgaacgagg
     1741 tttagagcaa gcttcaggaa actgagacag gaattttatt aaaaatttaa attttgaaga
     1801 aagttcaggg ttaatagcat ccattttttg ctttgcaagt tcctcagcat tcttaacaaa
     1861 agacgtctct tttgacatgt ttaaagttta aacctcctgt gtgaaattat tatccgctca
     1921 taattccaca cattatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     1981 tgagctaact cacattaatt gcgttgcgct cactgccaat tgctttccag tcgggaaacc
     2041 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     2101 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     2161 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     2221 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     2281 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     2341 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     2401 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     2461 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     2521 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     2581 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     2641 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     2701 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     2761 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     2821 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     2881 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     2941 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     3001 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     3061 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     3121 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     3181 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     3241 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     3301 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     3361 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     3421 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     3481 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     3541 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     3601 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     3661 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     3721 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     3781 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     3841 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     3901 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     3961 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     4021 cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa
     4081 ataggcgtat cacgaggcc
//
