LOCUS       sequence-446694-        2488 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ColA BBa_B0014 tetAp SD8 G1_PylRS-[mCNPR43] T3Te-T7Te DHFR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2488)
  AUTHORS   Costello A, Peterson AA, Lanster DL, Li Z, Carver GD, Badran AH
  TITLE     Efficient genetic code expansion without host genome modifications.
  JOURNAL   Nat Biotechnol. 2024 Sep 11. doi: 10.1038/s41587-024-02385-y.
  PUBMED    39261591
REFERENCE   2  (bases 1 to 2488)
  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"; journalName: "Nat 
            Biotechnol. 2024 Sep 11. doi: 10.1038/s41587-024-02385-y."
COMMENT     Sequence Label: pAB228i16
FEATURES             Location/Qualifiers
     source          1..2488
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(1..636)
                     /direction=LEFT
                     /label=ColA ori
                     /note="Plasmids containing the ColA origin of replication 
                     can be propagated in E. coli cells that contain additional 
                     plasmids with compatible origins."
     primer_bind     complement(146..165)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      659..686
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     promoter        788..843
                     /gene="tetR"
                     /label=tetR/tetA promoters
                     /note="overlapping promoters for bacterial tetR and tetA"
     protein_bind    794..812
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O1 for the tetR and tetA genes"
     protein_bind    824..842
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     terminator      1750..1779
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     terminator      complement(1780..1807)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
ORIGIN
        1 aaacgtccta gaagatgcca ggaggatact tagcagagag acaataaggc cggagcgaag
       61 ccgtttttcc ataggctccg cccccctgac gaacatcacg aaatctgacg ctcaaatcag
      121 tggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctga tggctccctc
      181 ttgcgctctc ctgttcccgt cctgcggcgt ccgtgttgtg gtggaggctt tacccaaatc
      241 accacgtccc gttccgtgta gacagttcgc tccaagctgg gctgtgtgca agaacccccc
      301 gttcagcccg actgctgcgc cttatccggt aactatcatc ttgagtccaa cccggaaaga
      361 cacgacaaaa cgccactggc agcagccatt ggtaactgag aattagtgga tttagatatc
      421 gagagtcttg aagtggtggc ctaacagagg ctacactgaa aggacagtat ttggtatctg
      481 cgctccacta aagccagtta ccaggttaag cagttcccca actgacttaa ccttcgatca
      541 aaccgcctcc ccaggcggtt ttttcgttta cagagcagga gattacgacg atcgtaaaag
      601 gatctcaaga agatcattta cggattcccg acaccaaaga ggacatccgg ttcacactgg
      661 ctcaccttcg ggtgggcctt tctgcgttta tatactagag agagaatata aaaagccaga
      721 ttattaatcc ggctttttta ttatttagac ttagggaccc tgcggccgcc attaattcct
      781 aatttttgtt gacactctat cattgataga gttattttac cactccctat cagtgataga
      841 gaaaagtgaa atgaatagac ctgcaggtgc agtaaggagg aaaaaaaaat ggttgttaaa
      901 ttcaccgatt ctcagattca gcatttgatg gaatacggtg ataacgattg gtctgaagcg
      961 gaattcgaag atgcggcggc gcgtgataaa gaattctctt ctcagttctc taaattgaaa
     1021 tctgcgaacg ataaaggttt gaaagatgtt attgcgaacc cgcgtaacga tttgaccgat
     1081 ttggaaaaca aaattcgtga aaaattggcg gcgcgtggtt tcattgaagt tcataccccg
     1141 attttcgttt ctaaatctgc gttggcgaaa atgaccatta ccgaagatca tccgttgttc
     1201 aaacaggttt tctggattga tgataaacgt gcgttgcgtc cgatgcatgc gatgaacgcg
     1261 ttcaaagtta tgcgtgaatt gcgtgatcat accaaaggtc cggttaaaat tttcgaaatt
     1321 ggttcttgtt tccgtaaaga atctaaatct tctacccatt tggaagaatt caccatgttg
     1381 aacttggttg aaatgggtcc ggatggtgat ccgatggaac atttgaaaat gtacattggt
     1441 gatattatgg atgcggttgg tgttgaatac accacctctc gtgaagaatc tgatgttttc
     1501 gttgaaacct tggatgttga aattaacggt accgaagttg cgtctggttc tgttggtccg
     1561 cataaattgg atccggcgca tgatgttcat gaaccgtacg cgggtattgg tttcggtttg
     1621 gaacgtttgt tgatgttgaa aaacggtaaa tctaacgcgc gtaaaaccgg taaatctatt
     1681 acctacttga acggttacaa attggattaa acttaattaa cggcactcct cagctgtaac
     1741 caactcactg gctcaccttc acgggtgagc ctttcttcgc gcagaaaggc ccacccgaag
     1801 gtgagccagt gtgattacat ttatgcccat ggcgtttaac ctttctgcca aatctggtaa
     1861 gagtagttaa tgttagacgc gaaatcctgg gtgaaaaccg gacggaagtt agacggaatt
     1921 tccgggaagt aaacatcacc ttccggttca atatcaatgg tagaaatatg caaggtatca
     1981 acctgatcaa tcaaagattt gtaaatttca ccaccaccag aaacaataac atgatcggta
     2041 atttttttca agttggtcaa cgcatcttta atagacggga aaatcaaaac gttttcgtta
     2101 tcagaggtga aagaagaacg ggtaacaacc gcgtatttac ggttcggcaa cgcacccata
     2161 gaaccgaagg ttttacgacc aaccaacaac cactggttgt aggtaatcgc tttgaacaac
     2221 aactgttcac ctttcgcaga ccacggaata tccggaccgt taccaataac accgttttta
     2281 gaaatcgcaa ccatcaaaga caatttcata attcttcctc agaggttaac attttattag
     2341 taagcatgct cgttttgtca cccgctgatg cttaccgtta attaatataa tcacttgtat
     2401 taaatagact ttctctaaaa atacaagaca ctctgttatt acaaatcgtg cgtgccgtct
     2461 atcctgcttg caaggccaaa taggccgt
//
