LOCUS       sequence-355025-        3703 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  M.BbrUI expression construct to improve transformation in B. breve 
            UCC2003.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3703)
  AUTHORS   Vento JM, Durmusoglu D, Li T, Patinios C, Sullivan S, Ttofali F, van
            Schaik J, Yu Y, Wang Y, Barquist L, Crook N, Beisel CL
  TITLE     A cell-free transcription-translation pipeline for recreating 
            methylation patterns boosts DNA transformation in bacteria.
  JOURNAL   Mol Cell. 2024 Jul 25;84(14):2785-2796.e4. doi: 
            10.1016/j.molcel.2024.06.003. Epub 2024 Jun 26.
  PUBMED    38936361
REFERENCE   2  (bases 1 to 3703)
  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"; doi: 
            "10.1016/j.molcel.2024.06.003"; journalName: "Mol Cell"; date: 
            "2024-07-25- 25"; volume: "84"; issue: "14"; pages: "2785-2796.e4"
COMMENT     Sequence Label: pJV457
FEATURES             Location/Qualifiers
     source          1..3703
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             345..367
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(1729..1746)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1900..2488)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1980..1999)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2659..3450,3451..3519))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     3282..3301
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3520..3624)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     join(3692..3703,1..7)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 acgaggccct ttcgtcttca agaattctgg cgaatcctct gaccagccag aaaacgacct
       61 ttctgtggtg aaaccggatg ctgcaattca gagcgccagc aagtggggga cagcagaaga
      121 cctgaccgcc gcagagtgga tgtttgacat ggtgaagact atcgcaccat cagccagaaa
      181 accgaatttt gctgggtggg ctaacgatat ccgcctgatg cgtgaacgtg acggacgtaa
      241 ccaccgcgac atgtgtgtgc tgttccgctg ggcatgctga gctaacaccg tgcgtgttga
      301 caattttacc tctggcggtg ataatggttg cagctagcaa taattttgtt taactttaag
      361 aaggagatat accatggacg ataatcgcac gcacgcggct accaacacag cacgcccgca
      421 agccctcgaa ttctttgcag gaataggcct agcacgcctt ggactcgaag aagccggctt
      481 ccaggtcgaa tggtcgaacg acatcgacgc cgccaagtgc gaaatgtatc gcaataactt
      541 cacggatacg ccgggccaca cccttgtcga gggggatatg agtgagctct ccggcgacga
      601 tctgccccac gatgcctcca tcgcttgggg atcctcccct tgcacggacc tctcccttgc
      661 aggcactcgt acgggcctga acggcaagca atccggcgct ttctggcaat acgtgcgact
      721 gctcgaagaa cttgggaaca cccggccgcc gattgccgtt ctcgagaatg tgaacggatt
      781 ggccacttct cactctgggg aagaccttgc cgcagcagtg catgccttca atgaactggg
      841 ctactccgta gacgtattgt ccatcgacac tcgtcgattt atcccccagt cccgaccgcg
      901 actcttttta gtgggcctgc agaatccgcc ggccgccgaa cagtttgatt cttccctgcg
      961 ccctgactgg ctgcagcagg tctttgcgga cacctctctc cgcacgcatc gcgcaccact
     1021 gcccgaaccg ccgtcccccc tgatgtctgg cctcaatatg tacgtcgaga gcatggcaga
     1081 cgacgacgaa cgctggtgga acgctgagag aactgccaaa ttccttgatt cgctgtcgcc
     1141 ggttcaggca actcgtgtag aaacgttgcg caatggggca accgattcct accggactgc
     1201 ttatcgccgc acccgcaacg gcaaagccgt gtgggaagtc cgcgcagacg acatctccgg
     1261 ctgcctacgc accgcacgag gcggctcttc tcgccaagcg atcattcgca tgggcaatgg
     1321 caaaaccgct gtccgttgga tgacaccacg cgaatacgct gctcttatgg gcgcagccca
     1381 ctacaacatt acgggtctac gccccaatca agtgatgttc ggattcggcg acgccgtctc
     1441 cgtccccgtc gtggaatggc ttggcaagca ctacttcatg cctgcactga acggcaccct
     1501 aggcaaagac ctatacgtgg agcaaactct gctctcaaat cacactgcat aactcgagca
     1561 aagcccgccg aaaggcgggc ttttctgtgt cgaccgatgc ccttgagagc cttcaaccca
     1621 gtcagctcct tccggtgggc gcggggcatg actatcgtcg ccgcacttat gactgtcttc
     1681 tttatcatgc aactcgtagg acaggtgccg gcagcgctct tccgcttcct cgctcactga
     1741 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     1801 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     1861 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     1921 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     1981 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     2041 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     2101 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     2161 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     2221 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     2281 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     2341 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     2401 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     2461 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     2521 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     2581 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     2641 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     2701 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     2761 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     2821 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     2881 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     2941 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     3001 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     3061 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     3121 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     3181 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     3241 tatgcggcga ccgagttgct cttgcccggc gtcaacacgg gataataccg cgccacatag
     3301 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     3361 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     3421 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     3481 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     3541 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     3601 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga
     3661 aaccattatt atcatgacat taacctataa aaataggcgt atc
//
