LOCUS       sequence-446696-        2494 bp    DNA     circular SYN 15-APR-2025
DEFINITION  ColA BBa_B0014 tetAp SD8 Lum1PylRS-[PylHRS*] T3Te-T7Te DHFR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2494)
  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 2494)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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: pAB228r3
FEATURES             Location/Qualifiers
     source          1..2494
                     /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      1756..1785
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     terminator      complement(1786..1813)
                     /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 ggatacccgt
      901 ttgaccccgg cgcaggcgca gcgtattcgt gaaatgggtg gtaccgttga tccgtctttg
      961 gcgttctctt ctgaagcgga acgtgaatct gcgttccagc gtatttctgc ggatttgcag
     1021 ggtgcgaact tggcgaaaat tcgtcgttgt gcggaagcgc cggaacgtca tccgattggt
     1081 tctttggaaa acaccttggc gtgtgcgttg gcggcgaaag gtttcattga agttaaaacc
     1141 ccgatgatga ttccggcgga tggtttggtt aaaatgggta ttgatgaatc tcatccgttg
     1201 tggaaccagg ttttctgggt tggtccgaaa aaagcgttgc gtccgatgtt ggcgccgaac
     1261 attttcttct tgggtcgtca tttgcgtcgt tctgttccgg cgccgttgtt gttgttcgaa
     1321 attggtccgt gtttccgtaa agaatctcgt ggttctaacc atttggaaga attcaccatg
     1381 ttgaacttgt tcgaattggc gccgcaggcg gatgcgaccg aacgtttgaa agaacatatt
     1441 gcgaccgtta tgaacgcggt tggtttgccg tacgaattgg ttgttgaagg ttctgaagtt
     1501 ttcggtacca ccattgatgt tgaagttgat ggtgttgaat tggcgtctgg tgcggttggt
     1561 ccgttgccga tggataaacc gcatggtatt accgaaccgt gggcgggtgt tggtttcggt
     1621 ttggaacgta ttgcgttgat gcgtaccaaa gaacagaaca ttaaaaaagt tggtcgttct
     1681 ttggtttacg ttaacggtgc gcgtattgat atttaaactt aattaacggc actcctcagc
     1741 tgtaaccaac tcactggctc accttcacgg gtgagccttt cttcgcgcag aaaggcccac
     1801 ccgaaggtga gccagtgtga ttacatttat gcccatggcg tttaaccttt ctgccaaatc
     1861 tggtaagagt agttaatgtt agacgcgaaa tcctgggtga aaaccggacg gaagttagac
     1921 ggaatttccg ggaagtaaac atcaccttcc ggttcaatat caatggtaga aatatgcaag
     1981 gtatcaacct gatcaatcaa agatttgtaa atttcaccac caccagaaac aataacatga
     2041 tcggtaattt ttttcaagtt ggtcaacgca tctttaatag acgggaaaat caaaacgttt
     2101 tcgttatcag aggtgaaaga agaacgggta acaaccgcgt atttacggtt cggcaacgca
     2161 cccatagaac cgaaggtttt acgaccaacc aacaaccact ggttgtaggt aatcgctttg
     2221 aacaacaact gttcaccttt cgcagaccac ggaatatccg gaccgttacc aataacaccg
     2281 tttttagaaa tcgcaaccat caaagacaat ttcataattc ttcctcagag gttaacattt
     2341 tattagtaag catgctcgtt ttgtcacccg ctgatgctta ccgttaatta atataatcac
     2401 ttgtattaaa tagactttct ctaaaaatac aagacactct gttattacaa atcgtgcgtg
     2461 ccgtctatcc tgcttgcaag gccaaatagg ccgt
//
