LOCUS       sequence-389657-        6318 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  E. coli machinery vector for the genetic encoding of acridone at TAG
            codons using the M. jannaschii Acd-A9 synthetase/tRNA pair.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6318)
  AUTHORS   Sungwienwong I, Hostetler ZM, Blizzard RJ, Porter JJ, Driggers CM, 
            Mbengi LZ, Villegas JA, Speight LC, Saven JG, Perona JJ, Kohli RM, 
            Mehl RA, Petersson EJ
  TITLE     Improving target amino acid selectivity in a permissive aminoacyl 
            tRNA synthetase through counter-selection.
  JOURNAL   Org Biomol Chem. 2017 May 3;15(17):3603-3610. doi: 
            10.1039/c7ob00582b.
  PUBMED    28397914
REFERENCE   2  (bases 1 to 6318)
  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"; journalName: 
            "Org Biomol Chem"; date: "2017-05-3- 3"; volume: "15"; issue: "17"; 
            pages: "3603-3610"
COMMENT     Sequence Label: pDule- Mj Acd A9
FEATURES             Location/Qualifiers
     source          1..6318
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1146..1175
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1191..1208
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1264..1281)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1434..1520
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(1727..1744)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1861..2406)
                     /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(1941..1960)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        join(2518..2523,2524..2540,2541..2546)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     CDS             2594..3784
                     /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(2946..2965)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     4091..4113
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4106..4122
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5106..5128
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5284..6162)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
ORIGIN
        1 aactcagaat aagaaatgag gccgctcatg gcgttctgtt gcccgtctca ccggtgaaaa
       61 gaaaaacaac cctggcgccg cttctttgag cgaacgatca aaaataagtg gcgccccatc
      121 aaaaaaatat tctcaacata aaaaactttg tgtaatactt gtaacgctga gtttacgctt
      181 tgaggaatcc cccatggacg aatttgaaat gataaagaga aacacatctg aaattatcag
      241 cgaggaagag ttaagagagg ttttaaaaaa agatgaaaaa tctgctgcga taggttttga
      301 accaagtggt aaaatacatt tagggcatta tctccaaata aaaaagatga ttgatttaca
      361 aaatgctgga tttgatataa ttatagattt ggctgattta cacgcctatt taaaccagaa
      421 aggagagttg gatgagatta gaaaaatagg agattataac aaaaaagttt ttgaagcaat
      481 ggggttaaag gcaaaatatg tttatggaag tgaactggag cttgataagg attatacact
      541 gaatgtctat agattggctt taaaaactac cttaaaaaga gcaagaagga gtatggaact
      601 tatagcaaga gaggatgaaa atccaaaggt tgctgaagtt atctatccaa taatgcaggt
      661 taattctatt cattatactg gcgttgatgt tgcagttgga gggatggagc agagaaaaat
      721 aaacatgtta gcaagggagc ttttaccaaa aaaggttgtt tgtattcaca accctgtctt
      781 aacgggtttg gatggagaag gaaagatgag ttcttcaaaa gggaatttta tagctgttga
      841 tgactctcca gaagagatta gggctaagat aaagaaagca tactgcccag ctggagttgt
      901 tgaaggaaat ccaataatgg agatagctaa atacttcctt gaatatcctt taaccataaa
      961 aaggccagaa aaatttggtg gagatttgac agttaatagc tatgaggagt tagagagttt
     1021 atttaaaaat aaggaattgc atccaatgga tttaaaaaat gctgtagctg aagaacttat
     1081 aaagatttta gagccaatta gaaagagatt ataaggtacc atatgggaat tcgaagcttg
     1141 ggcccgaaca aaaactcatc tcagaagagg atctgaatag cgccgtcgac catcatcatc
     1201 atcatcattg agtttaaacg gtctccagct tggctgtttt ggcggatgag agaagatttt
     1261 cagcctgata cagattaaat cagaacgcag aagcggtctg ataaaacaga atttgcctgg
     1321 cggcagtagc gcggtggtcc cacctgaccc catgccgaac tcagaagtga aacgccgtag
     1381 cgccgatggt agtgtggggt ctccccatgc gagagtaggg aactgccagg catcaaataa
     1441 aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg tcggtgaacg
     1501 ctctcctgag taggacaaat ccgccgggag ctgtccctcc tgttcagcta ctgacggggt
     1561 ggtgcgtaac ggcaaaagca ccgccggaca tcagcgctag cggagtgtat actggcttac
     1621 tatgttggca ctgatgaggg tgtcagtgaa gtgcttcatg tggcaggaga aaaaaggctg
     1681 caccggtgcg tcagcagaat atgtgataca ggatatattc cgcttcctcg ctcactgact
     1741 cgctacgctc ggtcgttcga ctgcggcgag cggaaatggc ttacgaacgg ggcggagatt
     1801 tcctggaaga tgccaggaag atacttaaca gggaagtgag agggccgcgg caaagccgtt
     1861 tttccatagg ctccgccccc ctgacaagca tcacgaaatc tgacgctcaa atcagtggtg
     1921 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggcggct ccctcgtgcg
     1981 ctctcctgtt cctgcctttc ggtttaccgg tgtcattccg ctgttatggc cgcgtttgtc
     2041 tcattccacg cctgacactc agttccgggt aggcagttcg ctccaagctg gactgtatgc
     2101 acgaaccccc cgttcagtcc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     2161 acccggaaag acatgcaaaa gcaccactgg cagcagccac tggtaattga tttagaggag
     2221 ttagtcttga agtcatgcgc cggttaaggc taaactgaaa ggacaagttt tggtgactgc
     2281 gctcctccaa gccagttacc tcggttcaaa gagttggtag ctcagagaac cttcgaaaaa
     2341 ccgccctgca aggcggtttt ttcgttttca gagcaagaga ttacgcgcag accaaaacga
     2401 tctcaagaag atcatcttat taatcagata aaatatttct agatttcagt gcaatttatc
     2461 tcttcaaatg tagcacctga agtcagcccc atacgatata agttgtaatt ctcatgtttg
     2521 acagcttatc atcgataagc tttaatgcgg tagtttatca cagttaaatt gctaacgcag
     2581 tcaggcaccg tgtatgaaat ctaacaatgc gctcatcgtc atcctcggca ccgtcaccct
     2641 ggatgctgta ggcataggct tggttatgcc ggtactgccg ggcctcttgc gggatatcgt
     2701 ccattccgac agcatcgcca gtcactatgg cgtgctgcta gcgctatatg cgttgatgca
     2761 atttctatgc gcacccgttc tcggagcact gtccgaccgc tttggccgcc gcccagtcct
     2821 gctcgcttcg ctacttggag ccactatcga ctacgcgatc atggcgacca cacccgtcct
     2881 gtggatcctc tacgccggac gcatcgtggc cggcatcacc ggcgccacag gtgcggttgc
     2941 tggcgcctat atcgccgaca tcaccgatgg ggaagatcgg gctcgccact tcgggctcat
     3001 gagcgcttgt ttcggcgtgg gtatggtggc aggccccgtg gccgggggac tgttgggcgc
     3061 catctccttg catgcaccat tccttgcggc ggcggtgctc aacggcctca acctactact
     3121 gggctgcttc ctaatgcagg agtcgcataa gggagagcgt cgaccgatgc ccttgagagc
     3181 cttcaaccca gtcagctcct tccggtgggc gcggggcatg actatcgtcg ccgcacttat
     3241 gactgtcttc tttatcatgc aactcgtagg acaggtgccg gcagcgctct gggtcatttt
     3301 cggcgaggac cgctttcgct ggagcgcgac gatgatcggc ctgtcgcttg cggtattcgg
     3361 aatcttgcac gccctcgctc aagccttcgt cactggtccc gccaccaaac gtttcggcga
     3421 gaagcaggcc attatcgccg gcatggcggc cgacgcgctg ggctacgtct tgctggcgtt
     3481 cgcgacgcga ggctggatgg ccttccccat tatgattctt ctcgcttccg gcggcatcgg
     3541 gatgcccgcg ttgcaggcca tgctgtccag gcaggtagat gacgaccatc agggacagct
     3601 tcaaggatcg ctcgcggctc ttaccagcct aacttcgatc attggaccgc tgatcgtcac
     3661 ggcgatttat gccgcctcgg cgagcacatg gaacgggttg gcatggattg taggcgccgc
     3721 cctatacctt gtctgcctcc ccgcgttgcg tcgcggtgca tggagccggg ccacctcgac
     3781 ctgaatggaa gccggcggca cctcgctaac ggattcacca ctccaagaat tggagccaat
     3841 caattcttgc ggagaactgt gaatgcgcaa accaaccctt ggcagaacat atccatcgcg
     3901 tccgccatct ccagcagccg cacgcggcgc atctcgggct ccttgcatgc accattcctt
     3961 gcggcggcgg tgctcaacgg cctcaaccta ctactgggct gcttcctaat gcaggagtcg
     4021 cataagggag agcgtctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg
     4081 ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgccaagct taaaaaaaat
     4141 ccttagcttt cgctaaggat ctgcagtggt ccggcgggcc ggatttgaac cagcgccatg
     4201 cggatttaga gtccgccgtt ctgccctgct gaactaccgc cgggaattca gcgttacaag
     4261 tattacacaa agttttttat gttgagaata tttttttgat ggggcgccac ttatttttga
     4321 tcgttcgctc aaagaagcgg cgccagggtt gtttttcttt tcaccagtga gacgggcaac
     4381 agaacgccat gagcggcctc atttcttatt ctgagttaca acagtccgca ccgctgccgg
     4441 tagctccttc cggtgggcgc ggggcatgac tatcgtcgcc gcacttatga ctgtcttctt
     4501 tatcatgcaa ctcgtaggac aggtgccggc agcgcccaac agtcccccgg ccacggggcc
     4561 tgccaccata cccacgccga aacaagcgcc ctgcaccatt atgttccgga tctgcatcgc
     4621 aggatgctgc tggctaccct gtggaacacc tacatctgta ttaacgaagc gctaaccgtt
     4681 tttatcatgc tctgggaggc agaataaatg atcatatcgt caattattac ctccacgggg
     4741 agagcctgag caaactggcc tcaggcattt gagaagcaca cggtcacact gcttccggta
     4801 gtcaataaac cggtaaacca gcaatagaca taagcggcta tttaacgacc ctgccctgaa
     4861 ccgacgaccg ggtcgaattt gctttcgaat ttctgccatt catccgctta ttatcactta
     4921 ttcaggcgta gcaaccaggc gtttaagggc accaataact gccttaaaaa aattacgccc
     4981 cgccctgcca ctcatcgcag ttgactgggt catggctgcg ccccgacacc cgccaacacc
     5041 cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac
     5101 cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca
     5161 gcagatcaat tcgcgcgcga aggcgaagcg gcatgcataa tgtgcctgtc aaatggacga
     5221 agcagggatt ctgcaaaccc tatgctactc cgtcaagccg tcaattgtct gattcgttac
     5281 caattatgac aacttgacgg ctacatcatt cactttttct tcacaaccgg cacggaactc
     5341 gctcgggctg gccccggtgc attttttaaa tacccgcgag aaatagagtt gatcgtcaaa
     5401 accaacattg cgaccgacgg tggcgatagg catccgggtg gtgctcaaaa gcagcttcgc
     5461 ctggctgata cgttggtcct cgcgccagct taagacgcta atccctaact gctggcggaa
     5521 aagatgtgac agacgcgacg gcgacaagca aacatgctgt gcgacgctgg cgatatcaaa
     5581 attgctgtct gccaggtgat cgctgatgta ctgacaagcc tcgcgtaccc gattatccat
     5641 cggtggatgg agcgactcgt taatcgcttc catgcgccgc agtaacaatt gctcaagcag
     5701 atttatcgcc agcagctccg aatagcgccc ttccccttgc ccggcgttaa tgatttgccc
     5761 aaacaggtcg ctgaaatgcg gctggtgcgc ttcatccggg cgaaagaacc ccgtattggc
     5821 aaatattgac ggccagttaa gccattcatg ccagtaggcg cgcggacgaa agtaaaccca
     5881 ctggtgatac cattcgcgag cctccggatg acgaccgtag tgatgaatct ctcctggcgg
     5941 gaacagcaaa atatcacccg gtcggcaaac aaattctcgt ccctgatttt tcaccacccc
     6001 ctgaccgcga atggtgagat tgagaatata acctttcatt cccagcggtc ggtcgataaa
     6061 aaaatcgaga taaccgttgg cctcaatcgg cgttaaaccc gccaccagat gggcattaaa
     6121 cgagtatccc ggcagcaggg gatcattttg cgcttcagcc atacttttca tactcccgcc
     6181 attcagagaa gaaaccaatt gtccatattg catcagacat tgccgtcact gcgtctttta
     6241 ctggctcttc tcgctaacca aaccggtaac ccgggaccaa gtttactcat atatacggac
     6301 agcggtgcgg actgttgt
//
