LOCUS       sequence-315316-        5064 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 5064)
  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 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: 
            "10.1021/acschembio.9b01026"; journalName: "ACS Chem Biol"; date: 
            "2020-02-21- 21"; volume: "15"; issue: "2"; pages: "562-574"
COMMENT     Sequence Label: pDule-Mb haloTyrRS C6
FEATURES             Location/Qualifiers
     source          1..5064
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     74..96
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     88..105
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     89..105
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             2193..2222
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2238..2255
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2311..2328)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2481..2567
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(2774..2791)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2908..3453)
                     /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(2988..3007)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        join(3565..3570,3571..3587,3588..3593)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     CDS             3641..4831
                     /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(3993..4012)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 tcgcataagg gagagcgtct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta
       61 acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa gcttaaaaaa
      121 aatccttagc tttcgctaag atctgcagtg gcggaaaccc cgggaatcta acccggctga
      181 acggatttag agtccattcg atctacatga tcaggttccc gcggccgcga attcagcgtt
      241 acaagtatta cacaaagttt tttatgttga gaatattttt ttgatggggc gccacttatt
      301 tttgatcgtt cgctcaaaga agcggcgcca gggttgtttt tcttttcacc agtgagacgg
      361 gcaacagaac gccatgagcg gcctcatttc ttattctgag ttacaacagt ccgcaccgct
      421 gccggtagct ccttccggtg ggcgcggggc atgactatcg tcgccgcact tatgactgtc
      481 ttctttatca tgcaactcgt aggacaggtg ccggcagcgc ccaacagtcc cccggccacg
      541 gggcctgcca ccatacccac gccgaaacaa gcgccctgca ccattatgtt ccggatctgc
      601 atcgcaggat gctgctggct accctgtgga acacctacgg tacctcgggt tgtcagcctg
      661 tcccgcttat aagatcatac gccgttatac gttgtttacg ctttgaggaa tcccatatga
      721 tggataaaaa accgctggat gtgctgatta gcgcggccgg cctgtggatg agccgtaccg
      781 gcaccctgca taaaatcaaa catcatgaag tgagccgcag caaaatctat attgaaatgg
      841 cgtgcggcga tcatctggtg gtgaacaaca gccgtagctg ccgtaccgcg cgtgcgtttc
      901 gtcatcataa ataccgcaaa acctgcaaac gttgccgtgt gagcgatgaa gatatcaaca
      961 acttactgac ccgtagcacc gaaagcaaaa acagcgtgaa agtgcgtgtg gtgagcgcgc
     1021 cgaaagtgaa aaaagcgatg ccgaaaagcg tgagccgtgc gccgaaaccg ctggaaaata
     1081 gcgtgagcgc gaaagcgagc accaacacca gccgtagcgt tccgagcccg gcgaaaagca
     1141 ccccgaacag cagcgttccg gcgtctgcgc cggcaccgag cctgacccgc agccagctgg
     1201 atcgtgtgga agcgctgctg tctccggaag ataaaattag cctgaacatg gcgaaaccgt
     1261 ttcgtgaact ggaaccggaa ctggtgaccc gtcgtaaaaa cgattttcag cgcctgtata
     1321 ccaacgatcg tgaagattat ctgggcaaac tggaacgtga tatcaccaaa ttttttgtgg
     1381 atcgcggctt tctggaaatt aaaagcccga ttctgattcc ggcggaatat gtggaacgta
     1441 tgggcattaa caacgacacc gaactgagca aacaaatttt ccgcgtggat aaaaacctgt
     1501 gcctgcgtcc gacgctggcc ccgacctcgc tgaactatct tcgtaaactg gatcgtattc
     1561 tgccgggtcc gatcaaaatt tttgaagtgg gcccgtgcta tcgcaaagaa agcgatggca
     1621 aagaacacct ggaagaattc accatggttg gttttactca aatgggcagc ggctgcaccc
     1681 gtgaaaacct ggaagcgctg atcaaagaat tcctggatta tctggaaatc gacttcgaaa
     1741 ttgtgggcga tagctgcatg gtgtatggcg ataccctgga tattatgcat ggcgatctgg
     1801 aactgagcag cgcggtggtg ggtccggtta gcctggatcg tgaatggggc attgataaac
     1861 cgtggattgg cgcgggtttt ggcctggaac gtctgctgaa agtgatgcat ggcttcaaaa
     1921 acattaaacg tgcgagccgt agcgaaagct actataacgg cattagcacg aacctgtaac
     1981 tgcagtttca aacgctaaat tgcctgatgc gctacgctta tcaggctaca tgatctctgc
     2041 aatatattga gtttgcgtgc ttttgtaggc cggataaggc gttcacgccg catccggcag
     2101 gaaacagcaa acaatccaaa acgccgcgtt cagcggcgtt ttttctgcgt ttcttcgcga
     2161 attaattccg ctctcgagcg aagcttgggc ccgaacaaaa actcatctca gaagaggatc
     2221 tgaatagcgc cgtcgaccat catcatcatc atcattgagt ttaaacggtc tccagcttgg
     2281 ctgttttggc ggatgagaga agattttcag cctgatacag attaaatcag aacgcagaag
     2341 cggtctgata aaacagaatt tgcctggcgg cagtagcgcg gtggtcccac ctgaccccat
     2401 gccgaactca gaagtgaaac gccgtagcgc cgatggtagt gtggggtctc cccatgcgag
     2461 agtagggaac tgccaggcat caaataaaac gaaaggctca gtcgaaagac tgggcctttc
     2521 gttttatctg ttgtttgtcg gtgaacgctc tcctgagtag gacaaatccg ccgggagctg
     2581 tccctcctgt tcagctactg acggggtggt gcgtaacggc aaaagcaccg ccggacatca
     2641 gcgctagcgg agtgtatact ggcttactat gttggcactg atgagggtgt cagtgaagtg
     2701 cttcatgtgg caggagaaaa aaggctgcac cggtgcgtca gcagaatatg tgatacagga
     2761 tatattccgc ttcctcgctc actgactcgc tacgctcggt cgttcgactg cggcgagcgg
     2821 aaatggctta cgaacggggc ggagatttcc tggaagatgc caggaagata cttaacaggg
     2881 aagtgagagg gccgcggcaa agccgttttt ccataggctc cgcccccctg acaagcatca
     2941 cgaaatctga cgctcaaatc agtggtggcg aaacccgaca ggactataaa gataccaggc
     3001 gtttccccct ggcggctccc tcgtgcgctc tcctgttcct gcctttcggt ttaccggtgt
     3061 cattccgctg ttatggccgc gtttgtctca ttccacgcct gacactcagt tccgggtagg
     3121 cagttcgctc caagctggac tgtatgcacg aaccccccgt tcagtccgac cgctgcgcct
     3181 tatccggtaa ctatcgtctt gagtccaacc cggaaagaca tgcaaaagca ccactggcag
     3241 cagccactgg taattgattt agaggagtta gtcttgaagt catgcgccgg ttaaggctaa
     3301 actgaaagga caagttttgg tgactgcgct cctccaagcc agttacctcg gttcaaagag
     3361 ttggtagctc agagaacctt cgaaaaaccg ccctgcaagg cggttttttc gttttcagag
     3421 caagagttta cgcgcagacc aaaacgatct caagaagatc atcttattaa tcagataaaa
     3481 tatttctaga tttcagtgca atttatctct tcaaatgtag cacctgaagt cagccccata
     3541 cgatataagt tgtaattctc atgtttgaca gcttatcatc gataagcttt aatgcggtag
     3601 tttatcacag ttaaattgct aacgcagtca ggcaccgtgt atgaaatcta acaatgcgct
     3661 catcgtcatc ctcggcaccg tcaccctgga tgctgtaggc ataggcttgg ttatgccggt
     3721 actgccgggc ctcttgcggg atatcgtcca ttccgacagc atcgccagtc actatggcgt
     3781 gctgctagcg ctatatgcgt tgatgcaatt tctatgcgca cccgttctcg gagcactgtc
     3841 cgaccgcttt ggccgccgcc cagtcctgct cgcttcgcta cttggagcca ctatcgacta
     3901 cgcgatcatg gcgaccacac ccgtcctgtg gatcctctac gccggacgca tcgtggccgg
     3961 catcaccggc gccacaggtg cggttgctgg cgcctatatc gccgacatca ccgatgggga
     4021 agatcgggct cgccacttcg ggctcatgag cgcttgtttc ggcgtgggta tggtggcagg
     4081 ccccgtggcc gggggactgt tgggcgccat ctccttgcat gcaccattcc ttgcggcggc
     4141 ggtgctcaac ggcctcaacc tactactggg ctgcttccta atgcaggagt cgcataaggg
     4201 agagcgtcga ccgatgccct tgagagcctt caacccagtc agctccttcc ggtgggcgcg
     4261 gggcatgact atcgtcgccg cacttatgac tgtcttcttt atcatgcaac tcgtaggaca
     4321 ggtgccggca gcgctctggg tcattttcgg cgaggaccgc tttcgctgga gcgcgacgat
     4381 gatcggcctg tcgcttgcgg tattcggaat cttgcacgcc ctcgctcaag ccttcgtcac
     4441 tggtcccgcc accaaacgtt tcggcgagaa gcaggccatt atcgccggca tggcggccga
     4501 cgcgctgggc tacgtcttgc tggcgttcgc gacgcgaggc tggatggcct tccccattat
     4561 gattcttctc gcttccggcg gcatcgggat gcccgcgttg caggccatgc tgtccaggca
     4621 ggtagatgac gaccatcagg gacagcttca aggatcgctc gcggctctta ccagcctaac
     4681 ttcgatcatt ggaccgctga tcgtcacggc gatttatgcc gcctcggcga gcacatggaa
     4741 cgggttggca tggattgtag gcgccgccct ataccttgtc tgcctccccg cgttgcgtcg
     4801 cggtgcatgg agccgggcca cctcgacctg aatggaagcc ggcggcacct cgctaacgga
     4861 ttcaccactc caagaattgg agccaatcaa ttcttgcgga gaactgtgaa tgcgcaaacc
     4921 aacccttggc agaacatatc catcgcgtcc gccatctcca gcagccgcac gcggcgcatc
     4981 tcgggctcct tgcatgcacc attccttgcg gcggcggtgc tcaacggcct caacctacta
     5041 ctgggctgct tcctaatgca ggag
//
