LOCUS       sequence-226941-        4011 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Chloramphenicol resistance cassette.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4011)
  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 4011)
  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 cat cassette
FEATURES             Location/Qualifiers
     source          1..4011
                     /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(1843..1849,1850..1867,1868..1873))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    1888..1909
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2029..2046)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2200..2788)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2280..2299)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2959..3750,3751..3819))
                     /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     3582..3601
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3820..3924)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3992..4010
                     /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 tgttacaggt ctctaataag caggtttcgc tacgctcaaa tcctttaaaa
      421 aaacacaaaa gaccacattt tttaatgtgg tcttttattc ttcaactaaa gcacccatta
      481 gttcaacaaa cgaaaattgg ataaagtggg atatttttaa aatatatatt tatgttacag
      541 taatattgac ttttaaaaaa ggattgattc taatgaagaa agcagacaag taagcctcct
      601 aaattcactt tagataaaaa tttaggaggc atatcaaatg aactttaata aaattgattt
      661 agacaattgg aagagaaaag agatatttaa tcattatttg aaccaacaaa cgacttttag
      721 tataaccaca gaaattgata ttagtgtttt ataccgaaac ataaaacaag aaggatataa
      781 attttaccct gcatttattt tcttagtgac aagggtgata aactcaaata cagcttttag
      841 aactggttac aatagcgacg gagagttagg ttattgggat aagttagagc cactttatac
      901 aatttttgat ggtgtatcta aaacattctc tggtatttgg actcctgtaa agaatgactt
      961 caaagagttt tatgatttat acctttctga tgtagagaaa tataatggtt cggggaaatt
     1021 gtttcccaaa acacctatac ctgaaaatgc tttttctctt tctattattc catggacttc
     1081 atttactggg tttaacttaa atatcaataa taatagtaat taccttctac ccattattac
     1141 agcaggaaaa ttcattaata aaggtaattc aatatattta ccgctatctt tacaggtaca
     1201 tcattctgtt tgtgatggtt atcatgcagg attgtttatg aactctattc aggaattgtc
     1261 agataggcct aatgactggc ttttataata tgagataatg ccgactgtac tttttacagt
     1321 cggttttcta atgtcactaa cctgccccgt tagttgaaga aggtttttat attacagctc
     1381 cagcggccgc atctaagaga cccaatgaat ctttctagaa gatctcctac aatattctca
     1441 gctgccatgg aaaatcgatg ttcttctttt attctctcaa gattttcagg ctgtatatta
     1501 aaacttatat taagaactat gctaaccacc tcatcaggaa ccgttgtagg tggcgtgggt
     1561 tttcttggca atcgactctc atgaaaacta cgagctaaat attcaatatg ttcctcttga
     1621 ccaactttat tctgcatttt ttttgaacga ggtttagagc aagcttcagg aaactgagac
     1681 aggaatttta ttaaaaattt aaattttgaa gaaagttcag ggttaatagc atccattttt
     1741 tgctttgcaa gttcctcagc attcttaaca aaagacgtct cttttgacat gtttaaagtt
     1801 taaacctcct gtgtgaaatt attatccgct cataattcca cacattatac gagccggaag
     1861 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     1921 ctcactgcca attgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     1981 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     2041 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     2101 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     2161 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     2221 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     2281 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     2341 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     2401 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     2461 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     2521 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     2581 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     2641 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     2701 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     2761 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     2821 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     2881 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     2941 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     3001 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     3061 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     3121 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     3181 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     3241 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     3301 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     3361 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     3421 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     3481 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     3541 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     3601 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     3661 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     3721 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     3781 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     3841 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     3901 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga
     3961 aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc c
//
