LOCUS       sequence-444961-        5581 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  RepA(N99D) SC101 T0-BS7 araC pBAD SD8 NpuC-C[AGGA]LFVY-NpuN-SsrA 
            ProK 5 tRNA operon tetA-orfL bla.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5581)
  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 5581)
  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: pAAP003l2
FEATURES             Location/Qualifiers
     source          1..5581
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      998..1220
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     promoter        2747..3031
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     2954..2973
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     protein_bind    3799..3815
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(5479..5550)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ttaaatacga ccgtatttcg ccaaaatgtt ttccaaggtg gtacgctgtt tctgggtcaa
       61 ccaagagaaa gaaccgttca aatcatgttt agactgcata tcagacaaga atttcgcttc
      121 gaaagaggtc aactgaattt tcgcggtcaa cgcatcatgc aaggttttac gcaaaccgtt
      181 acgcaagtaa gaatcagagg taatggtggt cggaatttta tcaccgttca ttttaatctg
      241 gttgttttcc aattcggtaa ccaaatccat ctgacgatcc aattcaacct ggaaaatcaa
      301 ggtatcggtc ggacgaccac gtttatcaac aaccaatttc atgttagagt aggtgttcaa
      361 atctttagaa atcggtttca aaacccactg gttcaaacgt ttgaattcat ggtagttgtt
      421 ttccaacatc aacatgaatt tgaattcatc caaagaaatt tcaatgttcg ctttatgggt
      481 ttttttctgg gtcaattctt tcaacaacca ttcgtaaata cgcatagagt atttgttttc
      541 gaaagattta acatgttcca agttgtattt aatgaatttt ttcaactgga acaagtacgg
      601 caaaatttct tcagagaaaa ccaattccaa tttttcagaa gagaatttcg cgtaatcggt
      661 ccactggaaa atttcgaaac ctttaaccaa cgggttacga atttcaacgg tacgggtcat
      721 caattcacgg gtcgctttcg ccaaaacacc gtacgcgttt tcacgagaaa tgttcatcat
      781 ctgcgcgtac tggttgtagg tgaaagaaac ggtacgttct ttacgggtcg ggttttcaat
      841 ggtcgggttc aacaacgcaa cacaacacaa aatcaatttg gtttcatgtt cggtcaaatc
      901 gtaacgagaa atcgccaatt cgttcgcttt gaaaacaacc aattcagaca tacatctcaa
      961 ttgtctaggt gattttaatc actataccaa ttgagatggg ctagtcaatg ataattacta
     1021 gtccttttcc tttgagttgt gggtatctgt aaattctgct agacctttgc tggaaaactt
     1081 gtaaattctg ctagaccctc tgtaaattcc gctagacctt tgtgtgtttt ttttgtttat
     1141 attcaagtgg ttataattta tagaataaag aaagaataaa aaaagataaa aagaatagat
     1201 cccagccctg tgtataactc actactttag tcagttccgc agtattacaa aaggatgtcg
     1261 caaacgctgt ttgctcctct acaaaacaga ccttaaaacc ctaaaggctt aagtagcacc
     1321 ctcgcaagct cgggcaaatc gctgaatatt ccttttgtct ccgaccatca ggcacctgag
     1381 tcgctgtctt tttcgtgaca ttcagttcgc tgcgctcacg gctctggcag tgaatggggg
     1441 taaatggcac tacaggcgcc ttttatggat tcatgcaagg aaactaccca taatacaaga
     1501 aaagcccgtc acgggcttct cagggcgttt tatggcgggt ctgctatgtg gtgctatctg
     1561 actttttgct gttcagcagt tcctgccctc tgattttcca gtctgaccac ttcggattat
     1621 cccgtgacag gtcattcaga ctggctaatg cacccagtaa ggcagcggta tcatcaacag
     1681 gcttacccgt cttactgtca agaggacatc cggtttgttc agaacgctcg gtcttgcaca
     1741 ccgggcgttt tttctttgtg agtccatagt acacttgaat aaataaaaac agccgttgcc
     1801 agaaagaggc acggctgttt tattttagac ttagggaccc tttaagacaa tttaaccgca
     1861 acatcgttaa ctttttcttc acaacccgca cggaattcag acggagacgc accggtacat
     1921 tttttgaaaa cacgagagaa gtacaactga tcatcgaaac caacgttacg accaacggtc
     1981 gcaatcggca tacgggtggt agacaacaac aatttcgcct gagaaatacg ctgatcttca
     2041 cgccaagaca aaacagaaat acccaactgc tgacggaaca aatgagacaa acgagacgga
     2101 gacaaacaaa catgctgcgc aacagacgca atatcgaagt tagaatccgc caaatgatca
     2161 gaaatgtact gacacgcttc acgaacacgg ttatccatcg gcggatgcaa agattcgtta
     2221 atcgcttcca tacgacgcaa caacaactgt tccaacaagt taatcgccaa caattcagag
     2281 taacgacctt caccctgacc cgcgttaata atctgaccga acaaatcaga gaaatgcggc
     2341 tgatgcgctt catccggacg gaagaaaccg gtgttcgcga aaatagacgg ccagttcaac
     2401 cattcatgcc agtacgcacg cggacggaag taaacccact gatggtacca ttcacgcgct
     2461 tccggatgac gaccgtaatg atgaatttca cccggcggga acaacaaaat atcacccgga
     2521 cgacaaacga attcacgacc ctggttttta acaacaccct gaccacgaat ggtcaagttc
     2581 aaaatgtaac ctttcatacc caacggacga tcaatgaaga aatccaagta accgttcgct
     2641 tcaatcgggg tcaaacccgc aaccaaatgc gcgttgaaag agtaacccgg caacaacgga
     2701 tcgttctgcg cttccgccat acttttcata ctcccaccat tcagagaaga aaccaattgt
     2761 ccatattgca tcagacattg ccgtcactgc gtcttttact ggctcttctc gctaacccaa
     2821 ccggtaaccc cgcttattaa aagcattctg taacaaagcg ggaccaaagc catgacaaaa
     2881 acgcgtaaca aaagtgtcta taatcacggc agaaaagtcc acattgatta tttgcacggc
     2941 gtcacacttt gctatgccat agcattttta tccataagat tagcggatcc tacctgacgc
     3001 tttttatcgc aactctctac tgtttctcca tacccgtttt acctgcaggt gcagtaagga
     3061 ggaaaaaaaa atgattaaaa ttgcgacccg taaatacttg ggtaaacaga acgtttacga
     3121 tattggtgtt gaacgtgatc ataacttcgc gttgaaaaac ggtttcattg cgtctaactg
     3181 taggattgtt cgtttactgt ttgtcttacg aaaccgaaat tttgaccgtt gaatacggtt
     3241 tgttgccgat tggtaaaatt gttgaaaaac gtattgaatg taccgtttac tctgttgata
     3301 acaacggtaa catttacacc cagccggttg cgcagtggca tgatcgtggt gaacaggaag
     3361 ttttcgaata ctgtttggaa gatggttctt tgattcgtgc gaccaaagat cataaattca
     3421 tgaccgttga tggtcagatg ttgccgattg atgaaatttt cgaacgtgaa ttggatttga
     3481 tgcgtgttga taacttgccg aacggtaccg cggcgaacga tgaaaactac gcgttggcgg
     3541 cgtaaactta attaacggca ctcaaataaa acgaaaggct cagtcgaaag actgggcctt
     3601 tcgttttgct gaggagactt agggaccctt gtgcttctca aatgcctgag gccagtttgc
     3661 tcaggctctc cccgtggagg taataattga cgatatgatc agtgcacggc taactaagcg
     3721 gcctgctgac tttctcgccg atcaaaaggc attttgctat taagggattg acgagggcgt
     3781 atctgcgcag taagataatt gtgagcggat aacaattagc agacaagata attttgcacc
     3841 cagcaaactt ggtacgtaaa cgcatcgtgg cgaactggtc cgggaccacc gagcattcct
     3901 aagctccggt tagccgggtt caactcccgg gttcgtcgcc aaaatttgaa agtgctgtaa
     3961 ggcacagacc acccaagggg gacggtccgg cgaccagcag gcctctataa gcctcgcctt
     4021 gcggggttcg acaccccggt ctctcgccag tttaaaagac atcggcgtca agcggatgtc
     4081 tggctgaaag gcctgaagaa tttggacctg tgatcatgta gatcgagagg cttctaatcc
     4141 atatcagccg ggttagattc ccggcgggtc cgccatctct tacttgatat ggctttagta
     4201 gcggtatcaa tatcagcagt aaaataaatt tcccgatccc gccctagctc agaggtagag
     4261 cgtgcctctc tagaagggca tggtccccgg ttcaaatcct gggggcggga ccaactactt
     4321 tatgtagtct ccgccgtgta gcaagaaatt gagaagtgaa ctggtggctc aatggtagag
     4381 ctgcacgctt ccgaacgtgt tggttgcagg ttcaagtcct gtccagttca ccaatttatt
     4441 caagacgctt accttgtaag tgcacccagt aggatccgtc gacctgcaga tccttagcga
     4501 aagctaagga ttttttttaa gcttctacca ttcatccgac ttaattaacg gcactcctca
     4561 gcaaatataa tgaccctctt gataacccaa gagggcattt tttaatgccc atggcgttta
     4621 ccaatgttta atcaaagacg caccaatttc cgcaatctga cggttacgtt catccatggt
     4681 cgcctgagaa ccggtggtgt aaataacaac aatacgagac ggtttaccat ccggacccaa
     4741 cgccgcaata ataccacgag aaccacgttc acccgcacca gatttatccg caatgaacca
     4801 acccgccggc aacgcagaac gcaacaacgg acccgcaact ttatccgctt ccatccaatc
     4861 aatcaactgc tgacgagacg ccaaggtcaa caattcaccg gtcaacaatt tacgcaaggt
     4921 ggtcgccatc gcaaccggca tggtggtatc acgttcatcg ttcggaatcg cttcgttcaa
     4981 ttccggttcc caacgatcca aacgggtaac atgatcaccc atgttatgca agaacgcggt
     5041 caattctttc ggaccaccaa tggtggtcaa caacaagttc gccgcggtgt tatcagacat
     5101 ggtaatcgcc gcagaacaca attcacgaac ggtcatacca tcggtcaaat gtttttcggt
     5161 aaccggagag tattcaacca aatcgttctg agagtaatga atacgacgac ccaactgttc
     5221 ctgacccgca tcaatacgag acaaaaccgc accacacaac aaaactttga aggtagacat
     5281 catcgggaaa cgttcttccg gacggaaaga ttccaaaatt ttaccagagt tcaaatccaa
     5341 ttcaatgtaa ccaacacgcg cacccaactg atcttccgca tctttaactt taaccaaggt
     5401 ttccggatgc gcgaaaaccg gcaaacagaa cgccgcgaag aacggaatca acgcaacacg
     5461 gaaatgctga atagacatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     5521 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaaggcc aaataagccg
     5581 t
//
