LOCUS       sequence-226774-        3607 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  smc gene upstream homology region.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3607)
  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 3607)
  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 'rncS
FEATURES             Location/Qualifiers
     source          1..3607
                     /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(1439..1445,1446..1463,1464..1469))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    1484..1505
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1625..1642)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1796..2384)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1876..1895)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2555..3346,3347..3415))
                     /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     3178..3197
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3416..3520)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3588..3606
                     /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 tgttacactc gagggtctcg cgagggatcc gacgaaattg agagccgcaa
      421 ttgtttgcga accgtctctc gtttcattgg ctcacgagct gtcattcggc gatcttgtcc
      481 tgttgggtaa aggcgaggaa atgacaggcg gaagaaagcg tcctgctcta ttggcggatg
      541 tttttgaggc atttatcgga gccttgtacc ttgaccaagg attagagccg gtcgaaagtt
      601 tcttaaaagt ttatgtgttc cctaaaatta acgatggtgc tttttctcat gtgatggatt
      661 tcaaaagcca gctgcaggaa tacgtgcagc gggacggcaa aggctctctg gagtataaaa
      721 tctccaacga aaaaggacct gcgcacaacc gtgaatttga agccatcgta tctctaaaag
      781 gtgaaccact cggagtcgga aacggccgtt caaagaaaga agccgaacag cacgctgctc
      841 aggaagcttt agctaaattg caaaaacacc atacgaaaca ataaaatccc ccttatgact
      901 cagggggatt tcagtatgta tgccgtctta tttgaccaat gtttatgata gaattgaaat
      961 acttattaca taaggaggat cgctatgttc cagagaccca atgaatcttt ctagaagatc
     1021 tcctacaata ttctcagctg ccatggaaaa tcgatgttct tcttttattc tctcaagatt
     1081 ttcaggctgt atattaaaac ttatattaag aactatgcta accacctcat caggaaccgt
     1141 tgtaggtggc gtgggttttc ttggcaatcg actctcatga aaactacgag ctaaatattc
     1201 aatatgttcc tcttgaccaa ctttattctg catttttttt gaacgaggtt tagagcaagc
     1261 ttcaggaaac tgagacagga attttattaa aaatttaaat tttgaagaaa gttcagggtt
     1321 aatagcatcc attttttgct ttgcaagttc ctcagcattc ttaacaaaag acgtctcttt
     1381 tgacatgttt aaagtttaaa cctcctgtgt gaaattatta tccgctcata attccacaca
     1441 ttatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     1501 cattaattgc gttgcgctca ctgccaattg ctttccagtc gggaaacctg tcgtgccagc
     1561 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     1621 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     1681 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     1741 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     1801 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     1861 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     1921 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     1981 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     2041 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     2101 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     2161 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     2221 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     2281 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     2341 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     2401 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     2461 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     2521 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     2581 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     2641 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     2701 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     2761 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     2821 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     2881 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     2941 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     3001 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     3061 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     3121 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     3181 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     3241 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     3301 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     3361 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     3421 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     3481 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     3541 cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca
     3601 cgaggcc
//
