LOCUS       sequence-345717-        5731 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Acridone82 tRNA synthetase and cognate amber suppressing tRNA 
            derived from M. barkeri for expression in E.coli..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5731)
  AUTHORS   Jones CM, Robkis DM, Blizzard RJ, Munari M, Venkatesh Y, Mihaila TS,
            Eddins AJ, Mehl RA, Zagotta WN, Gordon SE, Petersson EJ
  TITLE     Genetic encoding of a highly photostable, long lifetime fluorescent 
            amino acid for imaging in mammalian cells.
  JOURNAL   Chem Sci. 2021 Aug 3;12(36):11955-11964. doi: 10.1039/d1sc01914g. 
            eCollection 2021 Sep 22.
  PUBMED    34976337
REFERENCE   2  (bases 1 to 5731)
  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"; doi: 
            "10.1039/d1sc01914g"; journalName: "Chem Sci"; date: "2021-08-3- 3";
            volume: "12"; issue: "36"; pages: "11955-11964"
COMMENT     Sequence Label: pDule2-Acd82
FEATURES             Location/Qualifiers
     source          1..5731
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     69..86
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     CDS             complement(142..159)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(175..204)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(2289..2306)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2289..2305)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2298..2320)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3446..3465
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(3641..4432)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     promoter        complement(4433..4524)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4675..5220
                     /direction=RIGHT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     5121..5140
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5337..5354
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(5561..5647)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ggtcaggtgg gaccaccgcg ctactgccgc caggcaaatt ctgttttatc agaccgcttc
       61 tgcgttctga tttaatctgt atcaggctga aaatcttctc tcatccgcca aaacagccaa
      121 gctggagacc gtttaaactc aatgatgatg atgatgatgg tcgacggcgc tattcagatc
      181 ctcttctgag atgagttttt gttcgggccc aagcttcgct cgagagcgga attaattcgc
      241 gaagaaacgc agaaaaaacg ccgctgaacg cggcgttttg gattgtttgc tgtttcctgc
      301 cggatgcggc gtgaacgcct tatccggcct acaaaagcac gcaaactcaa tatattgcag
      361 agatcatgta gcctgataag cgtagcgcat caggcaattt agcgtttgaa actgcagtta
      421 caggttcgtg ctaatgccgt tatagtagct ttcgctacgg ctcgcacgtt taatgttttt
      481 gaagccatgc atcactttca gcagacgttc caggccaaaa cccgcgccaa tagtcggttt
      541 atcaatgccc cattcacgat ccaggctaac cggacccaca gccgcgctgc tcagttccag
      601 atcgccatgc ataatatcca gggtatcgcc atacaccatg cagctatcgc ccacaatttc
      661 gaagtcgatt tccagataat ccaggaattc tttgatcagc gcttccaggt tttcacgggt
      721 gcagccgctg cccatttgcc caaacgaaac catggtgaat tcttccaggt gttctttgcc
      781 atcgctttct ttgcgatagc acgggcccac ttcaaaaatt ttgatcggac ccggcagaat
      841 acgatccagt ttacgcagat agttatacag ggtcggggcc agcatcggac gcaggcacag
      901 gtttttatcc acgcggaaaa tttgtttgct cagttcggtg tcgttgttaa tgcccatacg
      961 ttccacatat tccgccggaa tcagaatcgg gcttttaatt tccagaaagc cgcgatccac
     1021 aaaaaatttg gtgatatcac gttccagttt gcccagataa tcttcacgat cgttggtata
     1081 caggcgctga aaatcgtttt tacgacgggt caccagttcc ggttccagtt cacgaaacgg
     1141 tttcgccatg ttcaggctaa ttttatcttc cggagacagc agcgcttcca cacgatccag
     1201 ctggctgcgg gtcaggctcg gtgccggcgc agacgccgga acgctgctgt tcggggtgct
     1261 tttcgccggg ctcggaacgc tacggctggt gttggtgctc gctttcgcgc tcacgctatt
     1321 ttccagcggt ttcggcgcac ggctcacgct tttcggcatc gcttttttca ctttcggcgc
     1381 gctcaccaca cgcactttca cgctgttttt gctttcggtg ctacgggtca gaaagttgtt
     1441 gatatcttca tcgctcacac ggcaacgttt gcaggttttg cggtatttat gatgacgaaa
     1501 cgcacgcgcg gtacggcagc tacggctgtt gttcaccacc agatgatcgc cgcacgccat
     1561 ttcaatatag attttgctgc ggctcacttc atgatgtttg attttatgca gggtgccggt
     1621 acggctcatc cacaggccgg tcgcgctaat cagcacatcc agcggttttt tatccatatg
     1681 ggattcctca aagcgtaaac aacgtataac ggcgtatgat cttataagcg ggacaggctg
     1741 acaacccgag gtaccgtagg tgttccacag ggtagccagc agcatcctgc gatgcagatc
     1801 cggaacataa tggtgcaggg cgcttgtttc ggcgtgggta tggtggcagg ccccgtggcc
     1861 gggggactgt tgggcgctgc cggcacctgt cctacgagtt gcatgataaa gaagacagtc
     1921 ataagtgcgg cgacgatagt catgccccgc gcccaccgga aggagctacc ggcagcggtg
     1981 cggactgttg taactcagaa taagaaatga ggccgctcat ggcgttctgt tgcccgtctc
     2041 actggtgaaa agaaaaacaa ccctggcgcc gcttctttga gcgaacgatc aaaaataagt
     2101 ggcgccccat caaaaaaata ttctcaacat aaaaaacttt gtgtaatact tgtaacgctg
     2161 aattcgcggc cgcgggaacc tgatcatgta gatcgaatgg actctaaatc cgttcagccg
     2221 ggttagattc ccggggtttc cgccactgca gatcttagcg aaagctaagg atttttttta
     2281 agcttggcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa
     2341 cttaatcgcc ttgcagcaca tccccctttc gccagacgct ctcccttatg cgactcctgc
     2401 attaggaagc agcccagtag taggttgagg ccgttgagca ccgccgccgc aaggaatggt
     2461 gcatgcaagg agcccgagat gcgccgcgtg cggctgctgg agatggcgga cgcgatggat
     2521 atgttctgcc aagggttggt ttgcgcattc acagttctcc gcaagaattg attggctcca
     2581 attcttggag tggtgaatcc gttagcgagg tgccgccggc ttccattcag gtcgaggtgg
     2641 cccggctcca tgcaccgcga cgcaacgcgg ggaggcagac aaggtatagg gcggcgccta
     2701 caatccatgc caacccgttc catgtgctcg ccgaggcggc ataaatcgcc gtgacgatca
     2761 gcggtccaat gatcgaagtt aggctggtaa gagccgcgag cgatccttga agctgtccct
     2821 gatggtcgtc atctacctgc ctggacagca tggcctgcaa cgcgggcatc ccgatgccgc
     2881 cggaagcgag aagaatcata atggggaagg ccatccagcc tcgcgtcgcg aacgccagca
     2941 agacgtagcc cagcgcgtcg gccgccatgc cggcgataat ggcctgcttc tcgccgaaac
     3001 gtttggtggc gggaccagtg acgaaggctt gagcgagggc gtgcaagatt ccgaataccg
     3061 caagcgacag gccgatcatc gtcgcgctcc agcgaaagcg gtcctcgccg aaaatgaccc
     3121 agagcgctgc cggcacctgt cctacgagtt gcatgataaa gaagacagtc ataagtgcgg
     3181 cgacgatagt catgccccgc gcccaccgga aggagctgac tgggttgaag gctctcaagg
     3241 gcatcggtcg acgctctccc ttatgcgact cctgcattag gaagcagccc agtagtaggt
     3301 tgaggccgtt gagcaccgcc gccgcaagga atggtgcatg caaggagatg gcgcccaaca
     3361 gtcccccggc cacggggcct gccaccatac ccacgccgaa acaagcgctc atgagcccga
     3421 agtggcgagc ccgatcttcc ccatcggtga tgtcggcgat ataggcgcca gcaaccgcac
     3481 ctgtggcgcc ggtgatgccg gccacgatgc gtccggcgta gaggatcccg gtaaaccagc
     3541 aatagacata agcggctatt taacgaccct gccctgaacc gacgaccggg tcatcgtggc
     3601 cggatcttgc ggcccctcgg cttgaacgaa ttgttagaca ttatttgccg actaccttgg
     3661 tgatctcgcc tttcacgtag tggacaaatt cttccaactg atctgcgcgc gaggccaagc
     3721 gatcttcttc ttgtccaaga taagcctgtc tagcttcaag tatgacgggc tgatactggg
     3781 ccggcaggcg ctccattgcc cagtcggcag cgacatcctt cggcgcgatt ttgccggtta
     3841 ctgcgctgta ccaaatgcgg gacaacgtaa gcactacatt tcgctcatcg ccagcccagt
     3901 cgggcggcga gttccatagc gttaaggttt catttagcgc ctcaaataga tcctgttcag
     3961 gaaccggatc aaagagttcc tccgccgctg gacctaccaa ggcaacgcta tgttctcttg
     4021 cttttgtcag caagatagcc agatcaatgt cgatcgtggc tggctcgaag atacctgcaa
     4081 gaatgtcatt gcgctgccat tctccaaatt gcagttcgcg cttagctgga taacgccacg
     4141 gaatgatgtc gtcgtgcaca acaatggtga cttctacagc gcggagaatc tcgctctctc
     4201 caggggaagc cgaagtttcc aaaaggtcgt tgatcaaagc tcgccgcgtt gtttcatcaa
     4261 gccttacggt caccgtaacc agcaaatcaa tatcactgtg tggcttcagg ccgccatcca
     4321 ctgcggagcc gtacaaatgt acggccagca acgtcggttc gagatggcgc tcgatgacgc
     4381 caactacctc tgatagttga gtcgatactt cggcgatcac cgcttccctc atactcttcc
     4441 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     4501 aatgtattta gaaaaataaa caaatagcta gctcactcgg tcgcatcgat gataagctgt
     4561 caaacatgag aattacaact tatatcgtat ggggctgact tcaggtgcta catttgaaga
     4621 gataaattgc actgaaatct agaaatattt tatctgatta ataagatgat cttcttgaga
     4681 tcgttttggt ctgcgcgtaa actcttgctc tgaaaacgaa aaaaccgcct tgcagggcgg
     4741 tttttcgaag gttctctgag ctaccaactc tttgaaccga ggtaactggc ttggaggagc
     4801 gcagtcacca aaacttgtcc tttcagttta gccttaaccg gcgcatgact tcaagactaa
     4861 ctcctctaaa tcaattacca gtggctgctg ccagtggtgc ttttgcatgt ctttccgggt
     4921 tggactcaag acgatagtta ccggataagg cgcagcggtc ggactgaacg gggggttcgt
     4981 gcatacagtc cagcttggag cgaactgcct acccggaact gagtgtcagg cgtggaatga
     5041 gacaaacgcg gccataacag cggaatgaca ccggtaaacc gaaaggcagg aacaggagag
     5101 cgcacgaggg agccgccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     5161 caccactgat ttgagcgtca gatttcgtga tgcttgtcag gggggcggag cctatggaaa
     5221 aacggctttg ccgcggccct ctcacttccc tgttaagtat cttcctggca tcttccagga
     5281 aatctccgcc ccgttcgtaa gccatttccg ctcgccgcag tcgaacgacc gagcgtagcg
     5341 agtcagtgag cgaggaagcg gaatatatcc tgtatcacat attctgctga cgcaccggtg
     5401 cagccttttt tctcctgcca catgaagcac ttcactgaca ccctcatcag tgccaacata
     5461 gtaagccagt atacactccg ctagcgctga tgtccggcgg tgcttttgcc gttacgcacc
     5521 accccgtcag tagctgaaca ggagggacag ctcccggcgg atttgtccta ctcaggagag
     5581 cgttcaccga caaacaacag ataaaacgaa aggcccagtc tttcgactga gcctttcgtt
     5641 ttatttgatg cctggcagtt ccctactctc gcatggggag accccacact accatcggcg
     5701 ctacggcgtt tcacttctga gttcggcatg g
//
