LOCUS       sequence-226903-        3873 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  smc downstream homology region.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3873)
  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 3873)
  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 sivC
FEATURES             Location/Qualifiers
     source          1..3873
                     /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(1705..1711,1712..1729,1730..1735))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    1750..1771
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1891..1908)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2062..2650)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2142..2161)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2821..3612,3613..3681))
                     /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     3444..3463
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3682..3786)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3854..3872
                     /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 tggtctctat ggccaagctt gcatgctaaa aaagaaaaag gcccgaagat
      421 gttcgggcct ttttcttacc tagggagaaa tcgtccaaat aataaagcga catatagctc
      481 ttctgttttg acgtcaacaa cgactgataa aaatcaaaca gataattcat tctcgttgtc
      541 ttttcgagtg acatttgagc tctccccttt gttaagtcaa atccctttac gataacttag
      601 tttacacggt tttaatgaca ctgtcaagtt tttatcttgt caaagcggcg attaactttt
      661 ccaaaatacg ctaaatatca tccaatttat tccgatattg tatatgaacc ctattttaag
      721 gaggattttt atgaaaaaaa tcggtttatt gtttatgtta tgtctggcag ccctgtttac
      781 aataggtttc ccggcacagc aagcagacgc tgctgaggca ccttataagg cctcaatcac
      841 aaatatcagc acagatggag gggtatacgg taaaatcaat tacggacaag gtcaatattg
      901 gcgagtcaag tacaatatta cagtaagcgg caaattactc gaccaaaacg gacagccagt
      961 gccaaacgcg cccgtaagat ttgaagcgga cacgaaggtc ggaaacacaa cacaaactgc
     1021 ttcaggaacg actgatgcaa acggaacatt tgaggttcca atgtatttgg ggccagctgc
     1081 tggttactat acgtattaca catctgtttc cgttcattac tatgacatca tcccattcag
     1141 agttttttct ggtgagtcaa gactggtatc cactgataac agcctttatc atttcgccta
     1201 tcaagtaaga agataaaaaa gaaaaaggcc ccaagatgtt ggggcctttt tcttaatcgt
     1261 agatgagacc atctttctag aagatctcct acaatattct cagctgccat ggaaaatcga
     1321 tgttcttctt ttattctctc aagattttca ggctgtatat taaaacttat attaagaact
     1381 atgctaacca cctcatcagg aaccgttgta ggtggcgtgg gttttcttgg caatcgactc
     1441 tcatgaaaac tacgagctaa atattcaata tgttcctctt gaccaacttt attctgcatt
     1501 ttttttgaac gaggtttaga gcaagcttca ggaaactgag acaggaattt tattaaaaat
     1561 ttaaattttg aagaaagttc agggttaata gcatccattt tttgctttgc aagttcctca
     1621 gcattcttaa caaaagacgt ctcttttgac atgtttaaag tttaaacctc ctgtgtgaaa
     1681 ttattatccg ctcataattc cacacattat acgagccgga agcataaagt gtaaagcctg
     1741 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc caattgcttt
     1801 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     1861 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     1921 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     1981 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     2041 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     2101 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     2161 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     2221 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     2281 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     2341 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     2401 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     2461 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     2521 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     2581 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     2641 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     2701 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     2761 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     2821 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     2881 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     2941 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     3001 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     3061 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     3121 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     3181 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     3241 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     3301 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     3361 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     3421 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     3481 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     3541 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     3601 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     3661 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     3721 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     3781 tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac
     3841 attaacctat aaaaataggc gtatcacgag gcc
//
