LOCUS       sequence-355029-        4490 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  M.SenLT2I expression construct to improve transformation in S. 
            enterica LT2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4490)
  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 4490)
  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: pJV303
FEATURES             Location/Qualifiers
     source          1..4490
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     557..576
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(795..899)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     967..985
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     RBS             1323..1345
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(3494..3511)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3665..4253)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3745..3764)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4424..4490,1..725,726..794))
                     /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"
ORIGIN
        1 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
       61 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
      121 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
      181 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
      241 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
      301 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
      361 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
      421 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
      481 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
      541 ccggcgtcaa cacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
      601 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
      661 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
      721 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
      781 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
      841 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
      901 acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc
      961 tataaaaata ggcgtatcac gaggcccttt cgtcttcaag aattctggcg aatcctctga
     1021 ccagccagaa aacgaccttt ctgtggtgaa accggatgct gcaattcaga gcgccagcaa
     1081 gtgggggaca gcagaagacc tgaccgccgc agagtggatg tttgacatgg tgaagactat
     1141 cgcaccatca gccagaaaac cgaattttgc tgggtgggct aacgatatcc gcctgatgcg
     1201 tgaacgtgac ggacgtaacc accgcgacat gtgtgtgctg ttccgctggg catgctgagc
     1261 taacaccgtg cgtgttgaca attttacctc tggcggtgat aatggttgca gctagcaata
     1321 attttgttta actttaagaa ggagatatac catggatgtt gaaagataac caaaaacaca
     1381 acgagtctgt tgccccgaat agcgcctttc tgtctgagtt acaacgtgca ttaccggaat
     1441 tttttaccgc cgatcgctat aacgagcagg gcgaactgat cgcgaaaggc ggatttgatc
     1501 tcgccagatt tgagcgcgcg ctgaaagcgc gtaatattga tgagctgact agcggttatc
     1561 agattgattt tattggcaaa gattacgcaa aaaaacaggc gggtgaaaaa tccgttaccg
     1621 ttatcgttcc tgacgtggaa cacaatactc tggcagaaaa taaaaacagc cataatcttt
     1681 ttctgaccgg agataatctg gatgtgttac gccatctgca aaataattac gccgataccg
     1741 tcgatatgat ctatatcgat cccccttata acaccggatc ggacgggttt gtctatcccg
     1801 atcattttga atatagcgat cgggcgttgc aggatatgtt tggtcttaat gataccgaac
     1861 tggcacgttt aaaatccatt cagggtaaat cgacgcactc cgcgtggtta tctttcatgt
     1921 atccgcgtct tttcctggcc aggaagctcc tgaaagatac cgggtttatt tttatctcta
     1981 tcgacgataa tgagtacgcc aatcttaaat taatgatgga tgagattttt ggcgaaggcg
     2041 gatttgtcac caatgtgatg tggaagcgca aaaaagagat ttctaacgac tctgataacg
     2101 tttccatcca gggggaatac attcttgttt atgccaaaac cggtcagggc gctttacgtt
     2161 tagaaccgct ttctaaagag tatattcaga aatcctataa agaaccgacc gaacagtttc
     2221 cagaagggaa atggcgaccg gtgccgttaa cggtgtcaaa agggctgagc ggcggcggct
     2281 atacctataa aattaccacg ccgaacggta cggtacacga aagactatgg gcttatcctg
     2341 aagccagtta ccaaaaactg gtggccgata atctggtcta ttttggcaaa gataacggcg
     2401 gtattcccca gcgagtcatg tacgcgcatc acagtaaggg gcagccaacg accaattact
     2461 gggataacgt agcgtcgaat aaagagggga aaaaggagat tctggatctc ttcggcgaca
     2521 acgtttttga tacgccgaaa ccgaccgcat tattgaagaa aatcatcaag ctcgctatcg
     2581 ataaagacgg cgtcgtcctg gacttttttg ccggttccgg cactacggcc catgcggtaa
     2641 tggcgctgaa tgaagaagat ggggggcagc gcacgtttat tctgtgtact atcgatcagg
     2701 cattaagcaa taacactatc gcgaaaaaag caggttataa cactattgat gaaatcagcc
     2761 gcgagcgaat tacacgcgtt gcggcgaaga tccgcgccaa caatcccgcg accaatagcg
     2821 atctcggttt taaacattat cgttttgcca ctccgacaca gcagacgctg gacgatctgg
     2881 atagcttcga tattgctacc ggccatttta tcaataccag cggtcaactg gccgctttca
     2941 ccgagtcagg atttaccgac atgatcaatc ctttttccgc cagaggattg ggcgtgccgg
     3001 gcggcgcaag cggcgaagag accttattaa cgacatggct ggtcgccgat ggttataaaa
     3061 tggatattga cgtacagacc gttgattttt ccggctattg cgccaggtat gttgataata
     3121 cgcgcctgta tctgattgat gaacgatggg gaacagagca gacccgcgat cttctcaacc
     3181 acattggtac gcaccagctt ccggttcaga ccattgtcat ttacggctac tctttcgacc
     3241 ttgaatccat tcgtgaactg gaaatcggct taaaacagct tgatcaaaaa gtgaacctgg
     3301 tgaagcgtta ttaataactc gagcaaagcc cgccgaaagg cgggcttttc tgtgtcgacc
     3361 gatgcccttg agagccttca acccagtcag ctccttccgg tgggcgcggg gcatgactat
     3421 cgtcgccgca cttatgactg tcttctttat catgcaactc gtaggacagg tgccggcagc
     3481 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     3541 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     3601 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     3661 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     3721 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     3781 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     3841 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     3901 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     3961 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     4021 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     4081 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     4141 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     4201 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     4261 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     4321 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     4381 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     4441 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
//
