LOCUS       sequence-355026-        5526 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MS.SenLT2II_L85Q 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 5526)
  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 5526)
  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: pJV505
FEATURES             Location/Qualifiers
     source          1..5526
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     561..580
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(799..903)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     971..989
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     RBS             1327..1349
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(4534..4551)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4705..5293)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4785..4804)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5464..5526,1..729,730..798))
                     /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 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
       61 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
      121 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
      181 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
      241 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
      301 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
      361 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
      421 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
      481 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
      541 ttgcccggcg tcaacacggg ataataccgc gccacatagc agaactttaa aagtgctcat
      601 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
      661 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
      721 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
      781 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
      841 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
      901 gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt
      961 aacctataaa aataggcgta tcacgaggcc ctttcgtctt caagaattct ggcgaatcct
     1021 ctgaccagcc agaaaacgac ctttctgtgg tgaaaccgga tgctgcaatt cagagcgcca
     1081 gcaagtgggg gacagcagaa gacctgaccg ccgcagagtg gatgtttgac atggtgaaga
     1141 ctatcgcacc atcagccaga aaaccgaatt ttgctgggtg ggctaacgat atccgcctga
     1201 tgcgtgaacg tgacggacgt aaccaccgcg acatgtgtgt gctgttccgc tgggcatgct
     1261 gagctaacac cgtgcgtgtt gacaatttta cctctggcgg tgataatggt tgcagctagc
     1321 aataattttg tttaacttta agaaggagat ataccatgaa caataacgat ctggtcgcca
     1381 agctgtggaa actgtgcgac aacctgcgtg acggcggcgt ctcctatcaa aactacgtca
     1441 atgaactcgc ctcgctgctg tttttgaaaa tgtgcaaaga gaccggccag gaagcggact
     1501 acctgccgga aggctaccgc tgggatgacc tgaaatcgcg tattggccag gatcagatgc
     1561 agttctaccg caacctgctg gttcagttag gctcagatga gaaaaagcag gtgcaggcgg
     1621 tattccataa tgtcagcacc accattgagc aacccaaaca gctgactgag ctggtcagct
     1681 atatggatgc gctggactgg tacaatggca accacgggaa atcccgcgac gacttcggcg
     1741 atatgtacga agggctgctg cagaagaacg ccaacgaaac caaatctggc gcgggccagt
     1801 acttcacccc acgcccgctg atcaaaacca ttatccatct gctgaaaccg cagccgcgtg
     1861 aagtggtgca ggacccggcg gcgggtacgg cgggcttttt gattgaagct gaccgttacg
     1921 tgaaatcaca gactaacgat ctggatgacc ttgatggcga cactcaggat ttccagatcc
     1981 accgcgcgtt tatcggcctc gaactggtgc ccggcacccg ccgtctggcg ctgatgaact
     2041 gtctgctgca cgatattgaa ggcaacctcg accacggcgg cgctatccgt ctgggcaaca
     2101 cgctgggcag cgacggtgaa aacctgccga aggcacatat cgtcgccacc aacccgccgt
     2161 ttggcagcgc cgcgggcacc aacattaccc gtaccttcgt gcacccgacc agcaacaaac
     2221 agctgtgctt tatgcagcat attattgaaa ccctgcaccc cggcggccgc gcggcggtgg
     2281 tggtgccgga taacgtgctg tttgaaggcg gtaagggcac cgatattcgt cgcgacctga
     2341 tggacaaatg ccatctgcac accatcctgc gtctgccgac cggtatcttt tacgcccagg
     2401 gcgtcaaaac caacgtgctg ttctttacca aaggcacggt caccaatccg catcaggata
     2461 aaaactgcac cgatgacgtg tgggtgtatg acctgcggac caatatgccg agcttcggca
     2521 agcgcacgcc gtttaccgag cagcatctgc agccgtttga aactgtctac ggcgaagatc
     2581 cacacggctt aagtccgcgt gaagaagggg agtggagctt taacgccgaa gagagcgaag
     2641 tcgccgacag cgaagagaac aagaacactg accagcacca ggccaccagc cgctggcgca
     2701 agttcagccg cgagtggatc cgcagcgcaa aatccgattc gctggatatc tcctggctga
     2761 aggataaaga cagcatcgac gccgacagtc tgccggagcc ggacgtgctg gcggcagaag
     2821 cgatgggcga gctggtacag gcgctgggcg aactggatgc gctgatgcgc gagctgggcg
     2881 cgggcgatga agcggatgct cagcgtcagt tgctgaatga agcgtttggc gaggtgaagg
     2941 catgagtggg gggaaattgc cggaggggtg ggctacaagc acaatcaatg aaatgtgcaa
     3001 cctcaatccc aaacttaaac ttgatgatga tttagatgtt ggatttatgc cgatggccgg
     3061 tgttccaaca acctatttag gtaaatgtaa ctttgaaacc aaaaagtgga gtgaagtaaa
     3121 aaaaggattt actcaatttc agaacgatga tgttattttt gccaaaatca cgccatgctt
     3181 cgaaaatgga aaagctgttg taatcaaaga attccctaac ggctatggtg ccggtagcac
     3241 tgaatattat gttcttcggt ctattaatgg gttaattaat ccccattggt tgtttgcttt
     3301 agttaaaact aaagattttt taactaatgg agcacttaat atgtccggtt cagtcggaca
     3361 taaacgtgtt actaaagaat ttcttgagaa ctatggtgtt cctgtcccac ctcttgccga
     3421 acaaaaagtc atcgccgaaa aactcgatac gctgctggcg caggtagaca gcaccaaagc
     3481 acgtcttgag caaatcccac aaatcctgaa acgttttcgc caatcagtga tagttgcagc
     3541 agtaaacggg caactgacaa aagaacttca taaaaaaaat aaattcaagt taacagaatt
     3601 gaatatttct attccatctt tatggaaaat cagtgagatt ggtcaatttg ctgatgtcaa
     3661 aggtggcaag agattaccta aaggtgaatc attaatagct gagaatacag ggtttccata
     3721 tattagagca gggcagctaa aaaatggaac tgttcttcct gaaggacaac tatacttaga
     3781 agaatatata caaaaaagta tttctagata tacagtctca tctggagatc tttatataac
     3841 tattgttggt gcatgtattg gtgatgctgg tataattcca gatgtttata ataatgcaaa
     3901 tttaactgaa aatgctgcca agatatgtaa tttaaacgaa aatattttca atagatttct
     3961 ttctttatgg ctgagaagta gttatcttca ggatattatc aattcagaaa taaaatcagg
     4021 agctcaaggg aagttggctt tggcaaggat aaaatcactc ccattgatac taccaccact
     4081 ccaagaacaa cacgaaatcg tccgccgcgt cgaacaactc ttcgcctacg ccgacaccat
     4141 tgaaaagcag gtcaacaacg ccctgacccg cgtcaacagc ctcacccagt cgatcctggc
     4201 gaaggccttc cgcggcgagc tgaccgccca gtggcgtgcg gaaaaccctg aacttatcag
     4261 cggtgaaaac agcgccgccg ccctgctgga aaaaattaag gccgaacgcg ccgccagcgg
     4321 cggtaaaaaa acctcgcgta aaaaagcctg aatctaactc gagcaaagcc cgccgaaagg
     4381 cgggcttttc tgtgtcgacc gatgcccttg agagccttca acccagtcag ctccttccgg
     4441 tgggcgcggg gcatgactat cgtcgccgca cttatgactg tcttctttat catgcaactc
     4501 gtaggacagg tgccggcagc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     4561 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     4621 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     4681 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     4741 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     4801 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     4861 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     4921 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     4981 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     5041 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     5101 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     5161 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     5221 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     5281 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     5341 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     5401 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     5461 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     5521 catagt
//
