LOCUS       sequence-444959-        2488 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ColA BBa_B0014 tetAp SD8 G1_PylRS-[pCNPR02] 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: pAB228i24
FEATURES             Location/Qualifiers
     source          1..2488
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(39..58)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      552..579
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     promoter        681..736
                     /gene="tetR"
                     /label=tetR/tetA promoters
                     /note="overlapping promoters for bacterial tetR and tetA"
     protein_bind    687..705
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O1 for the tetR and tetA genes"
     protein_bind    717..735
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     terminator      1643..1672
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     terminator      complement(1673..1700)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(join(2382..2488,1..529))
                     /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."
ORIGIN
        1 acgctcaaat cagtggtggc gaaacccgac aggactataa agataccagg cgtttccccc
       61 tgatggctcc ctcttgcgct ctcctgttcc cgtcctgcgg cgtccgtgtt gtggtggagg
      121 ctttacccaa atcaccacgt cccgttccgt gtagacagtt cgctccaagc tgggctgtgt
      181 gcaagaaccc cccgttcagc ccgactgctg cgccttatcc ggtaactatc atcttgagtc
      241 caacccggaa agacacgaca aaacgccact ggcagcagcc attggtaact gagaattagt
      301 ggatttagat atcgagagtc ttgaagtggt ggcctaacag aggctacact gaaaggacag
      361 tatttggtat ctgcgctcca ctaaagccag ttaccaggtt aagcagttcc ccaactgact
      421 taaccttcga tcaaaccgcc tccccaggcg gttttttcgt ttacagagca ggagattacg
      481 acgatcgtaa aaggatctca agaagatcat ttacggattc ccgacaccaa agaggacatc
      541 cggttcacac tggctcacct tcgggtgggc ctttctgcgt ttatatacta gagagagaat
      601 ataaaaagcc agattattaa tccggctttt ttattattta gacttaggga ccctgcggcc
      661 gccattaatt cctaattttt gttgacactc tatcattgat agagttattt taccactccc
      721 tatcagtgat agagaaaagt gaaatgaata gacctgcagg tgcagtaagg aggaaaaaaa
      781 aatggttgtt aaattcaccg attctcagat tcagcatttg atggaatacg gtgataacga
      841 ttggtctgaa gcggaattcg aagatgcggc ggcgcgtgat aaagaattct cttctcagtt
      901 ctctaaattg aaatctgcga acgataaagg tttgaaagat gttattgcga acccgcgtaa
      961 cgatttgacc gatttggaaa acaaaattcg tgaaaaattg gcggcgcgtg gtttcattga
     1021 agttcatacc ccgattttcg tttctaaatc tgcgttggcg aaaatgacca ttaccgaaga
     1081 tcatccgttg ttcaaacagg ttttctggat tgatgataaa cgtgcgttgc gtccgatgca
     1141 tgcgatgaac tctttcaaag ttatgcgtga attgcgtgat cataccaaag gtccggttaa
     1201 aattttcgaa attggttctt gtttccgtaa agaatctaaa tcttctaccc atttggaaga
     1261 attcaccatg ttgaacttgg ttgaaatggg tccggatggt gatccgatgg aacatttgaa
     1321 aatgtacatt ggtgatatta tggatgcggt tggtgttgaa tacaccacct ctcgtgaaga
     1381 atctgatgtt tgggttgaaa ccttggatgt tgaaattaac ggtaccgaag ttgcgtctgg
     1441 ttctgttggt ccgcataaat tggatccggc gcatgatgtt catgaaccgt acgcgggtat
     1501 tggtttcggt ttggaacgtt tgttgatgtt gaaaaacggt aaatctaacg cgcgtaaaac
     1561 cggtaaatct attacctact tgaacggtta caaattggat taaacttaat taacggcact
     1621 cctcagctgt aaccaactca ctggctcacc ttcacgggtg agcctttctt cgcgcagaaa
     1681 ggcccacccg aaggtgagcc agtgtgatta catttatgcc catggcgttt aacctttctg
     1741 ccaaatctgg taagagtagt taatgttaga cgcgaaatcc tgggtgaaaa ccggacggaa
     1801 gttagacgga atttccggga agtaaacatc accttccggt tcaatatcaa tggtagaaat
     1861 atgcaaggta tcaacctgat caatcaaaga tttgtaaatt tcaccaccac cagaaacaat
     1921 aacatgatcg gtaatttttt tcaagttggt caacgcatct ttaatagacg ggaaaatcaa
     1981 aacgttttcg ttatcagagg tgaaagaaga acgggtaaca accgcgtatt tacggttcgg
     2041 caacgcaccc atagaaccga aggttttacg accaaccaac aaccactggt tgtaggtaat
     2101 cgctttgaac aacaactgtt cacctttcgc agaccacgga atatccggac cgttaccaat
     2161 aacaccgttt ttagaaatcg caaccatcaa agacaatttc ataattcttc ctcagaggtt
     2221 aacattttat tagtaagcat gctcgttttg tcacccgctg atgcttaccg ttaattaata
     2281 taatcacttg tattaaatag actttctcta aaaatacaag acactctgtt attacaaatc
     2341 gtgcgtgccg tctatcctgc ttgcaaggcc aaataggccg taaacgtcct agaagatgcc
     2401 aggaggatac ttagcagaga gacaataagg ccggagcgaa gccgtttttc cataggctcc
     2461 gcccccctga cgaacatcac gaaatctg
//
