LOCUS       sequence-355028-        3928 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  M.BbrUIII 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 3928)
  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 3928)
  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: pJV461
FEATURES             Location/Qualifiers
     source          1..3928
                     /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(1954..1971)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2125..2713)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2205..2224)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2884..3675,3676..3744))
                     /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     3507..3526
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3745..3849)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     join(3917..3928,1..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 accatgagca agtccatcgc gcagatccgt aacgaggctg ctgccgctct
      421 ttccccggag cggaagcagc agtgggagca gtttctgacg cccgcccctg ttgccgagca
      481 ggcggccgga ctgttcactc ctgccgttga gcccgtgcgt atccttgatc tcggttcggg
      541 cacggggatc ctagctgcaa tggtcgccgc gcatgcgcca gccggcagtg gcgtcacagc
      601 catcgaacag gacaccgatc ttgccaaggc gtccgaggtt tcgatccgac aggtctgtga
      661 tgatgtcgag gtcatcgcca aatcggtgtt cgatgtgttg ctcgatgaca gattcgatcg
      721 cgtgattctc aacccgccgt ataagaagat cagtcccgta accattccca ctgccagcgg
      781 cggcgtgaag atcaccaacc tgtacaccgc ttttttggtg atcgctatcc aggcattggc
      841 cgaaagaggg gaatgcgtgg ccatcattcc tcgttcatgg atgaacggcg agtatttcaa
      901 ggacttccgc aaatggttgc tcgacgaatg cagccttgac gttctggccg tttacggctc
      961 gcggcaggac cacttcaagg acatggacat cctccaggag atcatgctgc tcaaggtgtc
     1021 caagcgaaag caaatcggtt caatcgccgt ttatgacggg atctccccgg agattccgtt
     1081 gggccggcag ccgcacgagc ttattccgtt ggaggcgctg accatgggta gggaccgcat
     1141 cctgcgcgtc gaacgtcagg atcctcggct ctcccgattc aagaccttgt ccgagcaagg
     1201 tatgtgggtg tccaccggca agctcgtgtg gttccgcaac cgcgacctgc tgtcggccac
     1261 cgagaagccg ggaagccacc ctttgtattg gtccgacaac cagcggggca tgacgaccgt
     1321 gcatccggtc aagtgcgaca gagaacagtg ggtcacgggc aaggcggatt cacgcaacgt
     1381 tgtgctgccc tcgggcagct actgcctggt caacaggttc tccgccaagg aacaacggca
     1441 ccgcatctac gcctcgtatc tgcgcagcga tacggagtac gtggcggaca acaagctgaa
     1501 ctacgtccac cagggcacct cccgccggac ggtgccgttg aacgacaagg tcgcgcgagg
     1561 cctgaccctt tggctgacgt caagcatcgt tgatgaatgg taccgccagg tgtcgggcag
     1621 cacacaggtc aacgccaccg acctacgcca gctgccgtgc ccatcaccgg ggcagctggc
     1681 agaactatcc gacctgctgc ccgtgaacga cgcggtggac cagcaaagca ttgaccatac
     1741 cttggaggga atgttctcat ggacaaaagc cagctaactc gagcaaagcc cgccgaaagg
     1801 cgggcttttc tgtgtcgacc gatgcccttg agagccttca acccagtcag ctccttccgg
     1861 tgggcgcggg gcatgactat cgtcgccgca cttatgactg tcttctttat catgcaactc
     1921 gtaggacagg tgccggcagc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     1981 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     2041 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     2101 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     2161 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     2221 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     2281 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     2341 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     2401 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     2461 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     2521 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     2581 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     2641 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     2701 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     2761 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     2821 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     2881 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     2941 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     3001 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     3061 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     3121 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     3181 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     3241 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     3301 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     3361 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     3421 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     3481 ttgctcttgc ccggcgtcaa cacgggataa taccgcgcca catagcagaa ctttaaaagt
     3541 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     3601 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     3661 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     3721 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     3781 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     3841 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat
     3901 gacattaacc tataaaaata ggcgtatc
//
