LOCUS       sequence-345712-        6318 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for incorporating the non-canonical amino acid 
            3-nitroTyrosine with the third generation Mj 3NY (A7) synthetase and
            cognate amber suppressing tRNA in Ecoli..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6318)
  AUTHORS   Beyer JN, Hosseinzadeh P, Gottfried-Lee I, Van Fossen EM, Zhu P, 
            Bednar RM, Karplus PA, Mehl RA, Cooley RB
  TITLE     Overcoming Near-Cognate Suppression in a Release Factor 1-Deficient 
            Host with an Improved Nitro-Tyrosine tRNA Synthetase.
  JOURNAL   J Mol Biol. 2020 Jul 24;432(16):4690-4704. doi: 
            10.1016/j.jmb.2020.06.014. Epub 2020 Jun 19.
  PUBMED    32569745
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: 
            "10.1016/j.jmb.2020.06.014"; journalName: "J Mol Biol"; date: 
            "2020-07-24- 24"; volume: "432"; issue: "16"; pages: "4690-4704"
COMMENT     Sequence Label: pDule-3-nitroTyrosine (A7)
FEATURES             Location/Qualifiers
     source          1..6318
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(564..580)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(573..595)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             complement(902..2092)
                     /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     1721..1740
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        complement(join(2140..2145,2146..2162,2163..2168))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     rep_origin      2280..2825
                     /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     2726..2745
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2942..2959
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3166..3252)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3405..3422
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     CDS             complement(3478..3495)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(3511..3540)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             4842..5720
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     primer_bind     complement(5876..5898)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 ataaaaacgg ttagcgcttc gttaatacag atgtaggtgt tccacagggt agccagcagc
       61 atcctgcgat gcagatccgg aacataatgg tgcagggcgc ttgtttcggc gtgggtatgg
      121 tggcaggccc cgtggccggg ggactgttgg gcgctgccgg cacctgtcct acgagttgca
      181 tgataaagaa gacagtcata agtgcggcga cgatagtcat gccccgcgcc caccggaagg
      241 agctaccggc agcggtgcgg actgttgtaa ctcagaataa gaaatgaggc cgctcatggc
      301 gttctgttgc ccgtctcact ggtgaaaaga aaaacaaccc tggcgccgct tctttgagcg
      361 aacgatcaaa aataagtggc gccccatcaa aaaaatattc tcaacataaa aaactttgtg
      421 taatacttgt aacgctgaat tcccggcggt agttcagcag ggcagaacgg cggactctaa
      481 atccgcatgg cgctggttca aatccggccc gccggaccac tgcagatcct tagcgaaagc
      541 taaggatttt ttttaagctt ggcactggcc gtcgttttac aacgtcgtga ctgggaaaac
      601 cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag acgctctccc
      661 ttatgcgact cctgcattag gaagcagccc agtagtaggt tgaggccgtt gagcaccgcc
      721 gccgcaagga atggtgcatg caaggagccc gagatgcgcc gcgtgcggct gctggagatg
      781 gcggacgcga tggatatgtt ctgccaaggg ttggtttgcg cattcacagt tctccgcaag
      841 aattgattgg ctccaattct tggagtggtg aatccgttag cgaggtgccg ccggcttcca
      901 ttcaggtcga ggtggcccgg ctccatgcac cgcgacgcaa cgcggggagg cagacaaggt
      961 atagggcggc gcctacaatc catgccaacc cgttccatgt gctcgccgag gcggcataaa
     1021 tcgccgtgac gatcagcggt ccaatgatcg aagttaggct ggtaagagcc gcgagcgatc
     1081 cttgaagctg tccctgatgg tcgtcatcta cctgcctgga cagcatggcc tgcaacgcgg
     1141 gcatcccgat gccgccggaa gcgagaagaa tcataatggg gaaggccatc cagcctcgcg
     1201 tcgcgaacgc cagcaagacg tagcccagcg cgtcggccgc catgccggcg ataatggcct
     1261 gcttctcgcc gaaacgtttg gtggcgggac cagtgacgaa ggcttgagcg agggcgtgca
     1321 agattccgaa taccgcaagc gacaggccga tcatcgtcgc gctccagcga aagcggtcct
     1381 cgccgaaaat gacccagagc gctgccggca cctgtcctac gagttgcatg ataaagaaga
     1441 cagtcataag tgcggcgacg atagtcatgc cccgcgccca ccggaaggag ctgactgggt
     1501 tgaaggctct caagggcatc ggtcgacgct ctcccttatg cgactcctgc attaggaagc
     1561 agcccagtag taggttgagg ccgttgagca ccgccgccgc aaggaatggt gcatgcaagg
     1621 agatggcgcc caacagtccc ccggccacgg ggcctgccac catacccacg ccgaaacaag
     1681 cgctcatgag cccgaagtgg cgagcccgat cttccccatc ggtgatgtcg gcgatatagg
     1741 cgccagcaac cgcacctgtg gcgccggtga tgccggccac gatgcgtccg gcgtagagga
     1801 tccacaggac gggtgtggtc gccatgatcg cgtagtcgat agtggctcca agtagcgaag
     1861 cgagcaggac tgggcggcgg ccaaagcggt cggacagtgc tccgagaacg ggtgcgcata
     1921 gaaattgcat caacgcatat agcgctagca gcacgccata gtgactggcg atgctgtcgg
     1981 aatggacgat atcccgcaag aggcccggca gtaccggcat aaccaagcct atgcctacag
     2041 catccagggt gacggtgccg aggatgacga tgagcgcatt gttagatttc atacacggtg
     2101 cctgactgcg ttagcaattt aactgtgata aactaccgca ttaaagctta tcgatgataa
     2161 gctgtcaaac atgagaatta caacttatat cgtatggggc tgacttcagg tgctacattt
     2221 gaagagataa attgcactga aatctagaaa tattttatct gattaataag atgatcttct
     2281 tgagatcgtt ttggtctgcg cgtaatctct tgctctgaaa acgaaaaaac cgccttgcag
     2341 ggcggttttt cgaaggttct ctgagctacc aactctttga accgaggtaa ctggcttgga
     2401 ggagcgcagt caccaaaact tgtcctttca gtttagcctt aaccggcgca tgacttcaag
     2461 actaactcct ctaaatcaat taccagtggc tgctgccagt ggtgcttttg catgtctttc
     2521 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcggact gaacgggggg
     2581 ttcgtgcata cagtccagct tggagcgaac tgcctacccg gaactgagtg tcaggcgtgg
     2641 aatgagacaa acgcggccat aacagcggaa tgacaccggt aaaccgaaag gcaggaacag
     2701 gagagcgcac gagggagccg ccagggggaa acgcctggta tctttatagt cctgtcgggt
     2761 ttcgccacca ctgatttgag cgtcagattt cgtgatgctt gtcagggggg cggagcctat
     2821 ggaaaaacgg ctttgccgcg gccctctcac ttccctgtta agtatcttcc tggcatcttc
     2881 caggaaatct ccgccccgtt cgtaagccat ttccgctcgc cgcagtcgaa cgaccgagcg
     2941 tagcgagtca gtgagcgagg aagcggaata tatcctgtat cacatattct gctgacgcac
     3001 cggtgcagcc ttttttctcc tgccacatga agcacttcac tgacaccctc atcagtgcca
     3061 acatagtaag ccagtataca ctccgctagc gctgatgtcc ggcggtgctt ttgccgttac
     3121 gcaccacccc gtcagtagct gaacaggagg gacagctccc ggcggatttg tcctactcag
     3181 gagagcgttc accgacaaac aacagataaa acgaaaggcc cagtctttcg actgagcctt
     3241 tcgttttatt tgatgcctgg cagttcccta ctctcgcatg gggagacccc acactaccat
     3301 cggcgctacg gcgtttcact tctgagttcg gcatggggtc aggtgggacc accgcgctac
     3361 tgccgccagg caaattctgt tttatcagac cgcttctgcg ttctgattta atctgtatca
     3421 ggctgaaaat cttctctcat ccgccaaaac agccaagctg gagaccgttt aaactcaatg
     3481 atgatgatga tgatggtcga cggcgctatt cagatcctct tctgagatga gtttttgttc
     3541 gggcccaagc ttcgaattcc catatggtac ctcataatct ctttctaatt ggctctaaaa
     3601 tctttataag ttcttcagct acagcatttt ttaaatccat tggatgcaat tccttatttt
     3661 taaataaact ctctaactcc tcatagctat taactgtcaa atctccacca aatttttctg
     3721 gcctttttat ggttaaagga tattcaagga agtatttagc tatctccatt attggatttc
     3781 cttcaacaac tccagctggg cagtatgctt tctttatctt agccctaatc tcttctggag
     3841 agtcatcaac agctataaaa ttcccttttg aagaactcat ctttccttct ccatccaaac
     3901 ccgttaagac agggttgtga atacaaacaa ccttttttgg taaaagctcc cttgctaaca
     3961 tgtgtatttt tctctgctcc atccctccaa ctgcaacatc aacgccacga taatgagcat
     4021 gattaacctg cattattgga tagataactt cagcaacctt tggattttca tcctctcttg
     4081 ctataagttc catactcctt cttgctcttt ttaaggtagt ttttaaagcc aatctataga
     4141 cattcagtgt ataatcctta tcaagctgga attcacttcc ataaacatat tttgccttta
     4201 accccattgc ttcaaaaact tttttgttat aatctcctat ttttctaatc tcatccaact
     4261 ctcctttctg gtttaaatag gccgttaaat cagccaaaag tataattata tcaaatccag
     4321 cattttgtaa atcaatcatc ttttttattt ggagataatg ccctaaatgt attttaccac
     4381 ttggttcaaa acctatatga gcagattttt catctttttt taaaacctct cttaactctt
     4441 cctcgctgat aatttcagat gtgtttctct ttatcatttc aaattcgtcc atgggggatt
     4501 cctcaaagcg taaactcagc gttacaagta ttacacaaag ttttttatgt tgagaatatt
     4561 tttttgatgg ggcgccactt atttttgatc gttcgctcaa agaagcggcg ccagggttgt
     4621 ttttcttttc accagtgaga cgggcaacag aacgccatga gcggcctcat ttcttattct
     4681 gagttacaac agtccgcacc gctgtccgta tatatgagta aacttggtcc cgggttaccg
     4741 gtttggttag cgagaagagc cagtaaaaga cgcagtgacg gcaatgtctg atgcaatatg
     4801 gacaattggt ttcttctctg aatggcggga gtatgaaaag tatggctgaa gcgcaaaatg
     4861 atcccctgct gccgggatac tcgtttaatg cccatctggt ggcgggttta acgccgattg
     4921 aggccaacgg ttatctcgat ttttttatcg accgaccgct gggaatgaaa ggttatattc
     4981 tcaatctcac cattcgcggt cagggggtgg tgaaaaatca gggacgagaa tttgtttgcc
     5041 gaccgggtga tattttgctg ttcccgccag gagagattca tcactacggt cgtcatccgg
     5101 aggctcgcga atggtatcac cagtgggttt actttcgtcc gcgcgcctac tggcatgaat
     5161 ggcttaactg gccgtcaata tttgccaata cggggttctt tcgcccggat gaagcgcacc
     5221 agccgcattt cagcgacctg tttgggcaaa tcattaacgc cgggcaaggg gaagggcgct
     5281 attcggagct gctggcgata aatctgcttg agcaattgtt actgcggcgc atggaagcga
     5341 ttaacgagtc gctccatcca ccgatggata atcgggtacg cgaggcttgt cagtacatca
     5401 gcgatcacct ggcagacagc aattttgata tcgccagcgt cgcacagcat gtttgcttgt
     5461 cgccgtcgcg tctgtcacat cttttccgcc agcagttagg gattagcgtc ttaagctggc
     5521 gcgaggacca acgtatcagc caggcgaagc tgcttttgag caccacccgg atgcctatcg
     5581 ccaccgtcgg tcgcaatgtt ggttttgacg atcaactcta tttctcgcgg gtatttaaaa
     5641 aatgcaccgg ggccagcccg agcgagttcc gtgccggttg tgaagaaaaa gtgaatgatg
     5701 tagccgtcaa gttgtcataa ttggtaacga atcagacaat tgacggcttg acggagtagc
     5761 atagggtttg cagaatccct gcttcgtcca tttgacaggc acattatgca tgccgcttcg
     5821 ccttcgcgcg cgaattgatc tgctgcctcg cgcgtttcgg tgatgacggt gaaaacctct
     5881 gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac
     5941 aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggcgcagcc atgacccagt
     6001 caactgcgat gagtggcagg gcggggcgta atttttttaa ggcagttatt ggtgccctta
     6061 aacgcctggt tgctacgcct gaataagtga taataagcgg atgaatggca gaaattcgaa
     6121 agcaaattcg acccggtcgt cggttcaggg cagggtcgtt aaatagccgc ttatgtctat
     6181 tgctggttta ccggtttatt gactaccgga agcagtgtga ccgtgtgctt ctcaaatgcc
     6241 tgaggccagt ttgctcaggc tctccccgtg gaggtaataa ttgacgatat gatcatttat
     6301 tctgcctccc agagcatg
//
