LOCUS       sequence-345716-        5064 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 5064)
  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 5064)
  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: pDule-Acd82
FEATURES             Location/Qualifiers
     source          1..5064
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             760..789
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             805..822
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(878..895)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1048..1134
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(1341..1358)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1475..2020)
                     /direction=LEFT
                     /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     complement(1555..1574)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        join(2132..2137,2138..2154,2155..2160)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     CDS             2208..3398
                     /codon_start=1
                     /gene="tet"
                     /product="tetracycline efflux protein"
                     /label=TcR
                     /note="confers resistance to tetracycline"
                     /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
                     GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
                     VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
                     LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
                     QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
                     AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
                     AALTSLTSIIGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
     primer_bind     complement(2560..2579)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     3705..3727
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3719..3736
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3720..3736
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gaacgtatgg gcattaacaa cgacaccgaa ctgagcaaac aaattttccg cgtggataaa
       61 aacctgtgcc tgcgtccgat gctggccccg accctgtata actatctgcg taaactggat
      121 cgtattctgc cgggtccgat caaaattttt gaagtgggcc cgtgctatcg caaagaaagc
      181 gatggcaaag aacacctgga agaattcacc atggtttcgt ttgggcaaat gggcagcggc
      241 tgcacccgtg aaaacctgga agcgctgatc aaagaattcc tggattatct ggaaatcgac
      301 ttcgaaattg tgggcgatag ctgcatggtg tatggcgata ccctggatat tatgcatggc
      361 gatctggaac tgagcagcgc ggctgtgggt ccggttagcc tggatcgtga atggggcatt
      421 gataaaccga ctattggcgc gggttttggc ctggaacgtc tgctgaaagt gatgcatggc
      481 ttcaaaaaca ttaaacgtgc gagccgtagc gaaagctact ataacggcat tagcacgaac
      541 ctgtaactgc agtttcaaac gctaaattgc ctgatgcgct acgcttatca ggctacatga
      601 tctctgcaat atattgagtt tgcgtgcttt tgtaggccgg ataaggcgtt cacgccgcat
      661 ccggcaggaa acagcaaaca atccaaaacg ccgcgttcag cggcgttttt tctgcgtttc
      721 ttcgcgaatt aattccgctc tcgagcgaag cttgggcccg aacaaaaact catctcagaa
      781 gaggatctga atagcgccgt cgaccatcat catcatcatc attgagttta aacggtctcc
      841 agcttggctg ttttggcgga tgagagaaga ttttcagcct gatacagatt aaatcagaac
      901 gcagaagcgg tctgataaaa cagaatttgc ctggcggcag tagcgcggtg gtcccacctg
      961 accccatgcc gaactcagaa gtgaaacgcc gtagcgccga tggtagtgtg gggtctcccc
     1021 atgcgagagt agggaactgc caggcatcaa ataaaacgaa aggctcagtc gaaagactgg
     1081 gcctttcgtt ttatctgttg tttgtcggtg aacgctctcc tgagtaggac aaatccgccg
     1141 ggagctgtcc ctcctgttca gctactgacg gggtggtgcg taacggcaaa agcaccgccg
     1201 gacatcagcg ctagcggagt gtatactggc ttactatgtt ggcactgatg agggtgtcag
     1261 tgaagtgctt catgtggcag gagaaaaaag gctgcaccgg tgcgtcagca gaatatgtga
     1321 tacaggatat attccgcttc ctcgctcact gactcgctac gctcggtcgt tcgactgcgg
     1381 cgagcggaaa tggcttacga acggggcgga gatttcctgg aagatgccag gaagatactt
     1441 aacagggaag tgagagggcc gcggcaaagc cgtttttcca taggctccgc ccccctgaca
     1501 agcatcacga aatctgacgc tcaaatcagt ggtggcgaaa cccgacagga ctataaagat
     1561 accaggcgtt tccccctggc ggctccctcg tgcgctctcc tgttcctgcc tttcggttta
     1621 ccggtgtcat tccgctgtta tggccgcgtt tgtctcattc cacgcctgac actcagttcc
     1681 gggtaggcag ttcgctccaa gctggactgt atgcacgaac cccccgttca gtccgaccgc
     1741 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg aaagacatgc aaaagcacca
     1801 ctggcagcag ccactggtaa ttgatttaga ggagttagtc ttgaagtcat gcgccggtta
     1861 aggctaaact gaaaggacaa gttttggtga ctgcgctcct ccaagccagt tacctcggtt
     1921 caaagagttg gtagctcaga gaaccttcga aaaaccgccc tgcaaggcgg ttttttcgtt
     1981 ttcagagcaa gagtttacgc gcagaccaaa acgatctcaa gaagatcatc ttattaatca
     2041 gataaaatat ttctagattt cagtgcaatt tatctcttca aatgtagcac ctgaagtcag
     2101 ccccatacga tataagttgt aattctcatg tttgacagct tatcatcgat aagctttaat
     2161 gcggtagttt atcacagtta aattgctaac gcagtcaggc accgtgtatg aaatctaaca
     2221 atgcgctcat cgtcatcctc ggcaccgtca ccctggatgc tgtaggcata ggcttggtta
     2281 tgccggtact gccgggcctc ttgcgggata tcgtccattc cgacagcatc gccagtcact
     2341 atggcgtgct gctagcgcta tatgcgttga tgcaatttct atgcgcaccc gttctcggag
     2401 cactgtccga ccgctttggc cgccgcccag tcctgctcgc ttcgctactt ggagccacta
     2461 tcgactacgc gatcatggcg accacacccg tcctgtggat cctctacgcc ggacgcatcg
     2521 tggccggcat caccggcgcc acaggtgcgg ttgctggcgc ctatatcgcc gacatcaccg
     2581 atggggaaga tcgggctcgc cacttcgggc tcatgagcgc ttgtttcggc gtgggtatgg
     2641 tggcaggccc cgtggccggg ggactgttgg gcgccatctc cttgcatgca ccattccttg
     2701 cggcggcggt gctcaacggc ctcaacctac tactgggctg cttcctaatg caggagtcgc
     2761 ataagggaga gcgtcgaccg atgcccttga gagccttcaa cccagtcagc tccttccggt
     2821 gggcgcgggg catgactatc gtcgccgcac ttatgactgt cttctttatc atgcaactcg
     2881 taggacaggt gccggcagcg ctctgggtca ttttcggcga ggaccgcttt cgctggagcg
     2941 cgacgatgat cggcctgtcg cttgcggtat tcggaatctt gcacgccctc gctcaagcct
     3001 tcgtcactgg tcccgccacc aaacgtttcg gcgagaagca ggccattatc gccggcatgg
     3061 cggccgacgc gctgggctac gtcttgctgg cgttcgcgac gcgaggctgg atggccttcc
     3121 ccattatgat tcttctcgct tccggcggca tcgggatgcc cgcgttgcag gccatgctgt
     3181 ccaggcaggt agatgacgac catcagggac agcttcaagg atcgctcgcg gctcttacca
     3241 gcctaacttc gatcattgga ccgctgatcg tcacggcgat ttatgccgcc tcggcgagca
     3301 catggaacgg gttggcatgg attgtaggcg ccgccctata ccttgtctgc ctccccgcgt
     3361 tgcgtcgcgg tgcatggagc cgggccacct cgacctgaat ggaagccggc ggcacctcgc
     3421 taacggattc accactccaa gaattggagc caatcaattc ttgcggagaa ctgtgaatgc
     3481 gcaaaccaac ccttggcaga acatatccat cgcgtccgcc atctccagca gccgcacgcg
     3541 gcgcatctcg ggctccttgc atgcaccatt ccttgcggcg gcggtgctca acggcctcaa
     3601 cctactactg ggctgcttcc taatgcagga gtcgcataag ggagagcgtc tggcgaaagg
     3661 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
     3721 taaaacgacg gccagtgcca agcttaaaaa aaatccttag ctttcgctaa gatctgcagt
     3781 ggcggaaacc ccgggaatct aacccggctg aacggattta gagtccattc gatctacatg
     3841 atcaggttcc cgcggccgcg aattcagcgt tacaagtatt acacaaagtt ttttatgttg
     3901 agaatatttt tttgatgggg cgccacttat ttttgatcgt tcgctcaaag aagcggcgcc
     3961 agggttgttt ttcttttcac cagtgagacg ggcaacagaa cgccatgagc ggcctcattt
     4021 cttattctga gttacaacag tccgcaccgc tgccggtagc tccttccggt gggcgcgggg
     4081 catgactatc gtcgccgcac ttatgactgt cttctttatc atgcaactcg taggacaggt
     4141 gccggcagcg cccaacagtc ccccggccac ggggcctgcc accataccca cgccgaaaca
     4201 agcgccctgc accattatgt tccggatctg catcgcagga tgctgctggc taccctgtgg
     4261 aacacctacg gtacctcggg ttgtcagcct gtcccgctta taagatcata cgccgttata
     4321 cgttgtttac gctttgagga atcccatatg atggataaaa aaccgctgga tgtgctgatt
     4381 agcgcgaccg gcctgtggat gagccgtacc ggcaccctgc ataaaatcaa acatcatgaa
     4441 gtgagccgca gcaaaatcta tattgaaatg gcgtgcggcg atcatctggt ggtgaacaac
     4501 agccgtagct gccgtaccgc gcgtgcgttt cgtcatcata aataccgcaa aacctgcaaa
     4561 cgttgccgtg tgagcgatga agatatcaac aactttctga cccgtagcac cgaaagcaaa
     4621 aacagcgtga aagtgcgtgt ggtgagcgcg ccgaaagtga aaaaagcgat gccgaaaagc
     4681 gtgagccgtg cgccgaaacc gctggaaaat agcgtgagcg cgaaagcgag caccaacacc
     4741 agccgtagcg ttccgagccc ggcgaaaagc accccgaaca gcagcgttcc ggcgtctgcg
     4801 ccggcaccga gcctgacccg cagccagctg gatcgtgtgg aagcgctgct gtctccggaa
     4861 gataaaatta gcctgaacat ggcgaaaccg tttcgtgaac tggaaccgga actggtgacc
     4921 cgtcgtaaaa acgattttca gcgcctgtat accaacgatc gtgaagatta tctgggcaaa
     4981 ctggaacgtg atatcaccaa attttttgtg gatcgcggct ttctggaaat taaaagcccg
     5041 attctgattc cggcggaata tgtg
//
