LOCUS       sequence-315318-        5734 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  C6 HaloTyrosine tRNA synthatase and cognate amber suppressing tRNA 
            derived from M. barkeri..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5734)
  AUTHORS   Jang HS, Gu X, Cooley RB, Porter JJ, Henson RL, Willi T, DiDonato 
            JA, Hazen SL, Mehl RA
  TITLE     Efficient Site-Specific Prokaryotic and Eukaryotic Incorporation of 
            Halotyrosine Amino Acids into Proteins.
  JOURNAL   ACS Chem Biol. 2020 Feb 21;15(2):562-574. doi: 
            10.1021/acschembio.9b01026. Epub 2020 Feb 10.
  PUBMED    31994864
REFERENCE   2  (bases 1 to 5734)
  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: 
            "10.1021/acschembio.9b01026"; journalName: "ACS Chem Biol"; date: 
            "2020-02-21- 21"; volume: "15"; issue: "2"; pages: "562-574"
COMMENT     Sequence Label: pDule2-Mb haloTyrRS C6
FEATURES             Location/Qualifiers
     source          1..5734
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     404..426
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     418..435
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     419..435
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             2523..2552
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2568..2585
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2641..2658)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2811..2897
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(3104..3121)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3238..3783)
                     /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(3318..3337)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3934..4025
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4026..4817
                     /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"
     primer_bind     complement(4993..5012)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 atggaacggg ttggcatgga ttgtaggcgc cgccctatac cttgtctgcc tccccgcgtt
       61 gcgtcgcggt gcatggagcc gggccacctc gacctgaatg gaagccggcg gcacctcgct
      121 aacggattca ccactccaag aattggagcc aatcaattct tgcggagaac tgtgaatgcg
      181 caaaccaacc cttggcagaa catatccatc gcgtccgcca tctccagcag ccgcacgcgg
      241 cgcatctcgg gctccttgca tgcaccattc cttgcggcgg cggtgctcaa cggcctcaac
      301 ctactactgg gctgcttcct aatgcaggag tcgcataagg gagagcgtct ggcgaaaggg
      361 ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt
      421 aaaacgacgg ccagtgccaa gcttaaaaaa aatccttagc tttcgctaag atctgcagtg
      481 gcggaaaccc cgggaatcta acccggctga acggatttag agtccattcg atctacatga
      541 tcaggttccc gcggccgcga attcagcgtt acaagtatta cacaaagttt tttatgttga
      601 gaatattttt ttgatggggc gccacttatt tttgatcgtt cgctcaaaga agcggcgcca
      661 gggttgtttt tcttttcacc agtgagacgg gcaacagaac gccatgagcg gcctcatttc
      721 ttattctgag ttacaacagt ccgcaccgct gccggtagct ccttccggtg ggcgcggggc
      781 atgactatcg tcgccgcact tatgactgtc ttctttatca tgcaactcgt aggacaggtg
      841 ccggcagcgc ccaacagtcc cccggccacg gggcctgcca ccatacccac gccgaaacaa
      901 gcgccctgca ccattatgtt ccggatctgc atcgcaggat gctgctggct accctgtgga
      961 acacctacgg tacctcgggt tgtcagcctg tcccgcttat aagatcatac gccgttatac
     1021 gttgtttacg ctttgaggaa tcccatatga tggataaaaa accgctggat gtgctgatta
     1081 gcgcggccgg cctgtggatg agccgtaccg gcaccctgca taaaatcaaa catcatgaag
     1141 tgagccgcag caaaatctat attgaaatgg cgtgcggcga tcatctggtg gtgaacaaca
     1201 gccgtagctg ccgtaccgcg cgtgcgtttc gtcatcataa ataccgcaaa acctgcaaac
     1261 gttgccgtgt gagcgatgaa gatatcaaca acttactgac ccgtagcacc gaaagcaaaa
     1321 acagcgtgaa agtgcgtgtg gtgagcgcgc cgaaagtgaa aaaagcgatg ccgaaaagcg
     1381 tgagccgtgc gccgaaaccg ctggaaaata gcgtgagcgc gaaagcgagc accaacacca
     1441 gccgtagcgt tccgagcccg gcgaaaagca ccccgaacag cagcgttccg gcgtctgcgc
     1501 cggcaccgag cctgacccgc agccagctgg atcgtgtgga agcgctgctg tctccggaag
     1561 ataaaattag cctgaacatg gcgaaaccgt ttcgtgaact ggaaccggaa ctggtgaccc
     1621 gtcgtaaaaa cgattttcag cgcctgtata ccaacgatcg tgaagattat ctgggcaaac
     1681 tggaacgtga tatcaccaaa ttttttgtgg atcgcggctt tctggaaatt aaaagcccga
     1741 ttctgattcc ggcggaatat gtggaacgta tgggcattaa caacgacacc gaactgagca
     1801 aacaaatttt ccgcgtggat aaaaacctgt gcctgcgtcc gacgctggcc ccgacctcgc
     1861 tgaactatct tcgtaaactg gatcgtattc tgccgggtcc gatcaaaatt tttgaagtgg
     1921 gcccgtgcta tcgcaaagaa agcgatggca aagaacacct ggaagaattc accatggttg
     1981 gttttactca aatgggcagc ggctgcaccc gtgaaaacct ggaagcgctg atcaaagaat
     2041 tcctggatta tctggaaatc gacttcgaaa ttgtgggcga tagctgcatg gtgtatggcg
     2101 ataccctgga tattatgcat ggcgatctgg aactgagcag cgcggtggtg ggtccggtta
     2161 gcctggatcg tgaatggggc attgataaac cgtggattgg cgcgggtttt ggcctggaac
     2221 gtctgctgaa agtgatgcat ggcttcaaaa acattaaacg tgcgagccgt agcgaaagct
     2281 actataacgg cattagcacg aacctgtaac tgcagtttca aacgctaaat tgcctgatgc
     2341 gctacgctta tcaggctaca tgatctctgc aatatattga gtttgcgtgc ttttgtaggc
     2401 cggataaggc gttcacgccg catccggcag gaaacagcaa acaatccaaa acgccgcgtt
     2461 cagcggcgtt ttttctgcgt ttcttcgcga attaattccg ctctcgagcg aagcttgggc
     2521 ccgaacaaaa actcatctca gaagaggatc tgaatagcgc cgtcgaccat catcatcatc
     2581 atcattgagt ttaaacggtc tccagcttgg ctgttttggc ggatgagaga agattttcag
     2641 cctgatacag attaaatcag aacgcagaag cggtctgata aaacagaatt tgcctggcgg
     2701 cagtagcgcg gtggtcccac ctgaccccat gccgaactca gaagtgaaac gccgtagcgc
     2761 cgatggtagt gtggggtctc cccatgcgag agtagggaac tgccaggcat caaataaaac
     2821 gaaaggctca gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc
     2881 tcctgagtag gacaaatccg ccgggagctg tccctcctgt tcagctactg acggggtggt
     2941 gcgtaacggc aaaagcaccg ccggacatca gcgctagcgg agtgtatact ggcttactat
     3001 gttggcactg atgagggtgt cagtgaagtg cttcatgtgg caggagaaaa aaggctgcac
     3061 cggtgcgtca gcagaatatg tgatacagga tatattccgc ttcctcgctc actgactcgc
     3121 tacgctcggt cgttcgactg cggcgagcgg aaatggctta cgaacggggc ggagatttcc
     3181 tggaagatgc caggaagata cttaacaggg aagtgagagg gccgcggcaa agccgttttt
     3241 ccataggctc cgcccccctg acaagcatca cgaaatctga cgctcaaatc agtggtggcg
     3301 aaacccgaca ggactataaa gataccaggc gtttccccct ggcggctccc tcgtgcgctc
     3361 tcctgttcct gcctttcggt ttaccggtgt cattccgctg ttatggccgc gtttgtctca
     3421 ttccacgcct gacactcagt tccgggtagg cagttcgctc caagctggac tgtatgcacg
     3481 aaccccccgt tcagtccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     3541 cggaaagaca tgcaaaagca ccactggcag cagccactgg taattgattt agaggagtta
     3601 gtcttgaagt catgcgccgg ttaaggctaa actgaaagga caagttttgg tgactgcgct
     3661 cctccaagcc agttacctcg gttcaaagag ttggtagctc agagaacctt cgaaaaaccg
     3721 ccctgcaagg cggttttttc gttttcagag caagagttta cgcgcagacc aaaacgatct
     3781 caagaagatc atcttattaa tcagataaaa tatttctaga tttcagtgca atttatctct
     3841 tcaaatgtag cacctgaagt cagccccata cgatataagt tgtaattctc atgtttgaca
     3901 gcttatcatc gatgcgaccg agtgagctag ctatttgttt atttttctaa atacattcaa
     3961 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     4021 agagtatgag ggaagcggtg atcgccgaag tatcgactca actatcagag gtagttggcg
     4081 tcatcgagcg ccatctcgaa ccgacgttgc tggccgtaca tttgtacggc tccgcagtgg
     4141 atggcggcct gaagccacac agtgatattg atttgctggt tacggtgacc gtaaggcttg
     4201 atgaaacaac gcggcgagct ttgatcaacg accttttgga aacttcggct tcccctggag
     4261 agagcgagat tctccgcgct gtagaagtca ccattgttgt gcacgacgac atcattccgt
     4321 ggcgttatcc agctaagcgc gaactgcaat ttggagaatg gcagcgcaat gacattcttg
     4381 caggtatctt cgagccagcc acgatcgaca ttgatctggc tatcttgctg acaaaagcaa
     4441 gagaacatag cgttgccttg gtaggtccag cggcggagga actctttgat ccggttcctg
     4501 aacaggatct atttgaggcg ctaaatgaaa ccttaacgct atggaactcg ccgcccgact
     4561 gggctggcga tgagcgaaat gtagtgctta cgttgtcccg catttggtac agcgcagtaa
     4621 ccggcaaaat cgcgccgaag gatgtcgctg ccgactgggc aatggagcgc ctgccggccc
     4681 agtatcagcc cgtcatactt gaagctagac aggcttatct tggacaagaa gaagatcgct
     4741 tggcctcgcg cgcagatcag ttggaagaat ttgtccacta cgtgaaaggc gagatcacca
     4801 aggtagtcgg caaataatgt ctaacaattc gttcaagccg aggggccgca agatccggcc
     4861 acgatgaccc ggtcgtcggt tcagggcagg gtcgttaaat agccgcttat gtctattgct
     4921 ggtttaccgg gatcctctac gccggacgca tcgtggccgg catcaccggc gccacaggtg
     4981 cggttgctgg cgcctatatc gccgacatca ccgatgggga agatcgggct cgccacttcg
     5041 ggctcatgag cgcttgtttc ggcgtgggta tggtggcagg ccccgtggcc gggggactgt
     5101 tgggcgccat ctccttgcat gcaccattcc ttgcggcggc ggtgctcaac ggcctcaacc
     5161 tactactggg ctgcttccta atgcaggagt cgcataaggg agagcgtcga ccgatgccct
     5221 tgagagcctt caacccagtc agctccttcc ggtgggcgcg gggcatgact atcgtcgccg
     5281 cacttatgac tgtcttcttt atcatgcaac tcgtaggaca ggtgccggca gcgctctggg
     5341 tcattttcgg cgaggaccgc tttcgctgga gcgcgacgat gatcggcctg tcgcttgcgg
     5401 tattcggaat cttgcacgcc ctcgctcaag ccttcgtcac tggtcccgcc accaaacgtt
     5461 tcggcgagaa gcaggccatt atcgccggca tggcggccga cgcgctgggc tacgtcttgc
     5521 tggcgttcgc gacgcgaggc tggatggcct tccccattat gattcttctc gcttccggcg
     5581 gcatcgggat gcccgcgttg caggccatgc tgtccaggca ggtagatgac gaccatcagg
     5641 gacagcttca aggatcgctc gcggctctta ccagcctaac ttcgatcatt ggaccgctga
     5701 tcgtcacggc gatttatgcc gcctcggcga gcac
//
