LOCUS       sequence-343331-        4449 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Recoded single aaRS/tRNA plasmid with MmPylRS and MmtRNA-Pyl-opt 
            with the ASL from the serT gene (UGA anticodon).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4449)
  AUTHORS   Robertson WE, Funke LFH, de la Torre D, Fredens J, Elliott TS, 
            Spinck M, Christova Y, Cervettini D, Boge FL, Liu KC, Buse S, Maslen
            S, Salmond GPC, Chin JW
  TITLE     Sense codon reassignment enables viral resistance and encoded 
            polymer synthesis.
  JOURNAL   Science. 2021 Jun 4;372(6546):1057-1062. doi: 
            10.1126/science.abg3029.
  PUBMED    34083482
REFERENCE   2  (bases 1 to 4449)
  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: "10.1126/science"; 
            journalName: "Science"; date: "2021-06-4- 4"; volume: "372"; issue: 
            "6546"; pages: "1057-1062"
COMMENT     Sequence Label: pMB1_MmPylRS_MmtRNA-Pyl-opt(UGA)
FEATURES             Location/Qualifiers
     source          1..4449
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(2144..2161)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2144..2160)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2153..2175)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             2505..2549
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(2587..2605)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2601..2648
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(2819..3634)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(2819..3634)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     3542..3561
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(3635..3739)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3790..4379)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3870..3889)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ggatcctcgg gagttgtcag cctgtcccgc ttataagatc atacgccgtt atacgttgtt
       61 tacgctttga ggaatcccat atgatggaca aaaaaccgct gaacaccctg atctctgcta
      121 ccggtctgtg gatgtctcgt accggtacca tccacaaaat caaacaccac gaagtttctc
      181 gttctaaaat ctacatcgaa atggcttgcg gtgaccacct ggttgttaac aactctcgtt
      241 cttctcgtac cgctcgtgct ctgcgtcacc acaaataccg taaaacctgc aaacgttgcc
      301 gtgtttctga cgaagacctg aacaaattcc tgaccaaagc taacgaagac cagacctctg
      361 ttaaagttaa agttgtttct gctccgaccc gtaccaaaaa agctatgccg aaatctgttg
      421 ctcgtgctcc gaaaccgctg gaaaacaccg aagctgctca ggctcagccg tctggttcta
      481 aattctctcc ggctatcccg gtttctaccc aggaatctgt ttctgttccg gcttctgttt
      541 ctacctctat ctcttctatc tctaccggtg ctaccgcttc tgctctggtt aaaggtaaca
      601 ccaacccgat cacctctatg tctgctccgg ttcaggcttc tgctccggct ctgaccaaat
      661 ctcagaccga ccgtctggaa gttctgctga acccgaaaga cgaaatctct ctgaactctg
      721 gtaaaccgtt ccgtgaactg gaatctgaac tgctgtctcg tcgtaaaaaa gacttacaac
      781 agatctacgc tgaagaacgt gaaaactacc tgggtaaact ggaacgtgaa atcacgcgtt
      841 tcttcgttga ccgtggtttc ctggaaatca aatctccgat cctgatcccg ctggaataca
      901 tcgaacgtat gggtatcgac aacgacaccg aactgtctaa acagatcttc cgtgttgaca
      961 aaaacttctg cctgcgtccg atgctggctc cgaacctgta caactacctg cgtaaactgg
     1021 accgtgctct gccggacccg atcaaaatct tcgaaatcgg tccgtgctac cgtaaagaat
     1081 ctgacggtaa agaacacctg gaagaattca ccatgctgaa cttctgccag atgggttctg
     1141 gttgcacccg tgaaaacctg gaatctatca tcaccgactt cctgaaccac ctgggtatcg
     1201 acttcaaaat cgttggtgac tcttgcatgg tttacggtga caccctggac gttatgcacg
     1261 gtgacctgga actgtcttct gctgttgttg gtccgatccc gctggaccgt gaatggggta
     1321 tcgacaaacc gtggatcggt gctggtttcg gtctggaacg tctgctgaaa gttaaacacg
     1381 acttcaaaaa catcaaacgt gctgctcgtt ctgaatctta ctacaacggt atctctacca
     1441 acctgtaact gcagtttcaa acgctaaatt gcctgatgcg ctacgcttat caggcctaca
     1501 tgatctctgc aatatattga gtttgcgtgc ttttgtaggc cggataaggc gttcacgccg
     1561 catccggcaa gaaacagcaa acaatccaaa acgccgcgtt cggcggtcga cacagatgta
     1621 ggtgttccac agggtagcca gcagcatcct gcgatgcaga tccggaacat aatggtgcag
     1681 ggcgcttgtt tcggcgtggg tatggtggca ggccccgtgg ccgggggact gttgggcgct
     1741 gccggcacct gtcctacgag ttgcatgata aagaagacag tcataagtgc ggcgacgata
     1801 gtcatgcccc gcgcccaccg gaaggagcta ccggcagcgg tgcggactgt tgtaactcag
     1861 aataagaaat gaggccgctc atggcgttct gttgcccgtc tcactggtga aaagaaaaac
     1921 aaccctggcg ccgcttcttt gagcgaacga tcaaaaataa gtggcgcccc atcaaaaaaa
     1981 tattctcaac ataaaaaact ttgtgtaata cttgtaacgc tagatctgga aacgtgatca
     2041 tgtagatcga acggtcttga aaaccgttca gtggggttag attccccacg tttccgccaa
     2101 ctagtatcct tagcgaaagc taaggatttt ttttaagctt ggcactggcc gtcgttttac
     2161 aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc
     2221 ctttcgccag acgctctccc ttatgcgact cctgcattag gaagcagccc agtagtaggt
     2281 tgaggccgtt gagcaccgcc gccgcaagga atggtgcatg caaggagccc gagatgcgcc
     2341 gcgtgcggct gctggagatg gcggacgcga tggatatgtt ctggcggccg cataatgctt
     2401 aagtcgaaca gaaagtaatc gtattgtaca cggccgcata atcgaaatta aattggatat
     2461 cggcaggcca cgcgatcgct gacgtcggta ccctcgagtc tggtaaagaa accgctgctg
     2521 cgaaatttga acgccagcac atggactcgt ctactagcgc agcttaatta acctaggctg
     2581 ctgccaccgc tgagcaataa ctagcataac cccttggggc ctctaaacgg gtcttgaggg
     2641 gttttttgct gaaacctcag gcatttgaga agcacacggt cacactgctt ccggtagtca
     2701 ataaaccggt aaaccagcaa tagacataag cggctattta acgaccctgc cctgaaccga
     2761 cgaccgggtc atcgtggccg gatcttgcgg cccctcggct tgaacgaatt gttagacatt
     2821 agaaaaactc atcgagcatc aaatgaaact gcaatttatt catatcagga ttatcaatac
     2881 catatttttg aaaaagccgt ttctgtaaac taggagaaaa ctcaccgagg cagttccata
     2941 ggatggcaag atcctggtat cggtctgcga ttccgactcg tccaacatca atacaaccta
     3001 ttaatttccc ctcgtcaaaa ataaggttat caagactgaa atcaccatga gtgacgacac
     3061 tatccggact gaatggcaaa agtttatgca tttctttcca gacttgttca acaggccagc
     3121 cattacgctc gtcatcaaaa tcactcgcat caaccaaacc gttattcatt cgactttgcg
     3181 cctgagcgag acgaaatacg cggtcgctgt taaaaggaca attacaaaca ggaatgctat
     3241 gcaaccggcg caggaacact gccagcgcat caacaatatt ttcaccacta tcaggatatt
     3301 cttctaatac ctggaatgct gttttcccgg ggatcgcagt ggtgagtaac catgcatcat
     3361 caggagtacg gataaaatgc ttgatggtcg gaagaggcat aaattccgtc agccagttta
     3421 gtctgaccat ctcatctgta acatcattgg caacgctacc tttgccatgt ttcagaaaca
     3481 actctggcgc atcgggcttc ccatacaatc gatagattgt cgcaccactt tgcccgacat
     3541 tatcgcgagc ccatttatac ccatataaat cagcatccat gttggaattt aatcgcggcc
     3601 tagagcaaga cgtttcccgt tgaatatggc tcatactctt cctttttcaa tattattgaa
     3661 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     3721 aacaaatagg ggttccgcga aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     3781 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     3841 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     3901 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     3961 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     4021 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     4081 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     4141 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     4201 gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa
     4261 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     4321 tagcggtggt tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     4381 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     4441 ggattttgg
//
