LOCUS       sequence-200689-        1196 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Recoded strain C321.Delta-A (MutS inactivated) with genomically 
            integrated Ubiquitin-UAG-sfGFP reporter and ClpS inactivated.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1196)
  AUTHORS   Kunjapur AM, Stork DA, Kuru E, Vargas-Rodriguez O, Landon M, Soll D,
            Church GM
  TITLE     Engineering posttranslational proofreading to discriminate 
            nonstandard amino acids.
  JOURNAL   Proc Natl Acad Sci U S A. 2018 Jan 16;115(3):619-624. doi: 
            10.1073/pnas.1715137115. Epub 2018 Jan 4.
  PUBMED    29301968
REFERENCE   2  (bases 1 to 1196)
  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: 
            "10.1073/pnas.1715137115"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2018-01-16- 16"; volume: "115"; issue: "3"; pages: "619-624"
COMMENT     Sequence Label: C321.Delta-ClpS.Ub-UAG-sfGFP
FEATURES             Location/Qualifiers
     source          1..1196
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        join(1..19,20..25,26..42,43..48,49..74)
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    1..19
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    26..44
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             84..95
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             102..329
                     /codon_start=1
                     /product="Saccharomyces cerevisiae ubiquitin"
                     /label=ubiquitin
                     /translation="MQIFVKTLTGKTITLEVESSDTIDNVKSKIQDKEGIPPDQQRLIF
                     AGKQLEDGRTLSDYNIQKESTLHLVLRLRGG"
     CDS             1071..1088
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      1133..1179
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 tccctatcag tgatagagat tgacatccct atcagtgata gagatactga gcacatcagc
       61 aggacgcact gaccgaattc attaaagagg agaaaggtac catgcagatt tttgtgaaga
      121 ctttaacagg taagacgatt accctggagg tggagtcctc ggacaccatc gataatgtaa
      181 aatcaaaaat ccaagataag gaaggaatcc ctccagacca gcaacgtctg attttcgcag
      241 gtaaacaact ggaggatggt cgcacgcttt cggactacaa catccagaaa gaatctaccc
      301 ttcatttggt tctgcgtctg cgtggaggat agttgtttgt gcaggagctt gcatccaagg
      361 gcgaggagct ctttactggc gtagtaccaa ttctcgtaga gctcgatggc gatgtaaatg
      421 gccataagtt ttccgtacgc ggcgagggcg agggcgatgc aactaacggc aagctcactc
      481 tcaagtttat ttgtactact ggcaagctcc cagtaccatg gccaactctc gtaactactc
      541 tgacctatgg cgtacaatgt ttttcccgct atccagatca catgaagcaa catgattttt
      601 ttaagtccgc aatgccagag ggctatgtac aagagcgcac tattagcttt aaggatgatg
      661 gcacctataa gactcgcgca gaggtaaagt ttgagggcga tactctcgta aatcgcattg
      721 agctcaaggg cattgatttt aaggaggatg gcaatattct cggccataag ctggagtata
      781 atttcaattc ccataatgta tacattaccg cagataagca aaagaatggc attaaggcga
      841 attttaagat tcgccataat gtggaggatg gctccgtaca actcgcagat cattatcaac
      901 aaaatactcc aattggcgat ggcccagtac tcctcccaga taatcattat ctctccactc
      961 aatccgtgct ctccaaagat ccaaatgaga agcgcgatca catggtactc ctggagtttg
     1021 taactgcagc aggcattact catggcatgg atgagctcta taagctcgag caccaccacc
     1081 accaccacta aaagcttgat gggggatccc atggtacgcg tgctagaggc atcaaataaa
     1141 acgaaaggct cagtcgaaag actgggcctt tcgttttatc tgttgtttgt cggtga
//
