LOCUS       sequence-444745-        3032 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ColA BBa_B0014 tetAp SD8 MmPylRS-[AF] T3Te-T7Te DHFR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3032)
  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 3032)
  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: pAB228p4
FEATURES             Location/Qualifiers
     source          1..3032
                     /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      2293..2322
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     terminator      complement(2323..2350)
                     /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 atgaataaac ctgcaggtgc agtaaggagg aaaaaaaaat ggataaaaaa
      901 ccgttgaaca ccttgatttc tgcgaccggt ttgtggatgt ctcgtaccgg taccattcat
      961 aaaattaaac atcatgaagt ttctcgttct aaaatttaca ttgaaatggc gtgtggtgat
     1021 catttggttg ttaacaactc tcgttcttct cgtaccgcgc gtgcgttgcg tcatcataaa
     1081 taccgtaaaa cctgtaaacg ttgtcgtgtt tctgatgaag atttgaacaa attcttgacc
     1141 aaagcgaacg aagatcagac ctctgttaaa gttaaagttg tttctgcgcc gacccgtacc
     1201 aaaaaagcga tgccgaaatc tgttgcgcgt gcgccgaaac cgttggaaaa caccgaagcg
     1261 gcgcaggcgc agccgtctgg ttctaaattc tctccggcga ttccggtttc tacccaggaa
     1321 tctgtttctg ttccggcgtc tgtttctacc tctatttctt ctatttctac cggtgcgacc
     1381 gcgtctgcgt tggttaaagg taacaccaac ccgattacct ctatgtctgc gccggttcag
     1441 gcgtctgcgc cggcgttgac caaatctcag accgatcgtt tggaagtttt gttgaacccg
     1501 aaagatgaaa tttctttgaa ctctggtaaa ccgttccgtg aattggaatc tgaattgttg
     1561 tctcgtcgta aaaaagattt gcagcagatt tacgcggaag aacgtgaaaa ctacttgggt
     1621 aaattggaac gtgaaattac ccgtttcttc gttgatcgtg gtttcttgga aattaaatct
     1681 ccgattttga ttccgttgga atacattgaa cgtatgggta ttgataacga taccgaattg
     1741 tctaaacaga ttttccgtgt tgataaaaac ttctgtttgc gtccgatgtt ggcgccgaac
     1801 ttggcgaact acttgcgtaa attggatcgt gcgttgccgg atccgattaa aattttcgaa
     1861 attggtccgt gttaccgtaa agaatctgat ggtaaagaac atttggaaga attcaccatg
     1921 ttgaacttct gtcagatggg ttctggttgt acccgtgaaa acttggaatc tattattacc
     1981 gatttcttga accatttggg tattgatttc aaaattgttg gtgattcttg tatggttttc
     2041 ggtgatacct tggatgttat gcatggtgat ttggaattgt cttctgcggt tgttggtccg
     2101 attccgttgg atcgtgaatg gggtattgat aaaccgtgga ttggtgcggg tttcggtttg
     2161 gaacgtttgt tgaaagttaa acatgatttc aaaaacatta aacgtgcggc gcgttctgaa
     2221 tcttactaca acggtatttc taccaacttg taaacttaat taacggcact cctcagctgt
     2281 aaccaactca ctggctcacc ttcacgggtg agcctttctt cgcgcagaaa ggcccacccg
     2341 aaggtgagcc agtgtgatta catttatgcc catggcgttt aacctttctg ccaaatctgg
     2401 taagagtagt taatgttaga cgcgaaatcc tgggtgaaaa ccggacggaa gttagacgga
     2461 atttccggga agtaaacatc accttccggt tcaatatcaa tggtagaaat atgcaaggta
     2521 tcaacctgat caatcaaaga tttgtaaatt tcaccaccac cagaaacaat aacatgatcg
     2581 gtaatttttt tcaagttggt caacgcatct ttaatagacg ggaaaatcaa aacgttttcg
     2641 ttatcagagg tgaaagaaga acgggtaaca accgcgtatt tacggttcgg caacgcaccc
     2701 atagaaccga aggttttacg accaaccaac aaccactggt tgtaggtaat cgctttgaac
     2761 aacaactgtt cacctttcgc agaccacgga atatccggac cgttaccaat aacaccgttt
     2821 ttagaaatcg caaccatcaa agacaatttc ataattcttc ctcagaggtt aacattttat
     2881 tagtaagcat gctcgttttg tcacccgctg atgcttaccg ttaattaata taatcacttg
     2941 tattaaatag actttctcta aaaatacaag acactctgtt attacaaatc gtgcgtgccg
     3001 tctatcctgc ttgcaaggcc aaatagggcc gt
//
