LOCUS       sequence-343328-        5193 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Recoded double aaRS/tRNA plasmid with 
            1R26PylRS/AlvtRNA-Delta-NPyl(8)(CGA).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5193)
  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 5193)
  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_1R26PylRS(CbzK)_AfTyrRS(p-I-Phe)_AlvtRNA-Delta-NPyl(8)(CGA)_Aft
            RNA-Tyr(A01)(CUA)
FEATURES             Location/Qualifiers
     source          1..5193
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             197..241
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(279..297)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      293..340
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(511..1326)
                     /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(511..1326)
                     /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     1234..1253
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(1327..1431)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1482..2071)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1562..1581)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     RBS             3085..3093
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     RBS             4117..4125
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     primer_bind     complement(5029..5046)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5029..5045)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5038..5060)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 gaatggtgca tgcaaggagc ccgagatgcg ccgcgtgcgg ctgctggaga tggcggacgc
       61 gatggatatg ttctggcggc cgcataatgc ttaagtcgaa cagaaagtaa tcgtattgta
      121 cacggccgca taatcgaaat taaattggat atcggcaggc cacgcgatcg ctgacgtcgg
      181 taccctcgag tctggtaaag aaaccgctgc tgcgaaattt gaacgccagc acatggactc
      241 gtctactagc gcagcttaat taacctaggc tgctgccacc gctgagcaat aactagcata
      301 accccttggg gcctctaaac gggtcttgag gggttttttg ctgaaacctc aggcatttga
      361 gaagcacacg gtcacactgc ttccggtagt caataaaccg gtaaaccagc aatagacata
      421 agcggctatt taacgaccct gccctgaacc gacgaccggg tcatcgtggc cggatcttgc
      481 ggcccctcgg cttgaacgaa ttgttagaca ttagaaaaac tcatcgagca tcaaatgaaa
      541 ctgcaattta ttcatatcag gattatcaat accatatttt tgaaaaagcc gtttctgtaa
      601 actaggagaa aactcaccga ggcagttcca taggatggca agatcctggt atcggtctgc
      661 gattccgact cgtccaacat caatacaacc tattaatttc ccctcgtcaa aaataaggtt
      721 atcaagactg aaatcaccat gagtgacgac actatccgga ctgaatggca aaagtttatg
      781 catttctttc cagacttgtt caacaggcca gccattacgc tcgtcatcaa aatcactcgc
      841 atcaaccaaa ccgttattca ttcgactttg cgcctgagcg agacgaaata cgcggtcgct
      901 gttaaaagga caattacaaa caggaatgct atgcaaccgg cgcaggaaca ctgccagcgc
      961 atcaacaata ttttcaccac tatcaggata ttcttctaat acctggaatg ctgttttccc
     1021 ggggatcgca gtggtgagta accatgcatc atcaggagta cggataaaat gcttgatggt
     1081 cggaagaggc ataaattccg tcagccagtt tagtctgacc atctcatctg taacatcatt
     1141 ggcaacgcta cctttgccat gtttcagaaa caactctggc gcatcgggct tcccatacaa
     1201 tcgatagatt gtcgcaccac tttgcccgac attatcgcga gcccatttat acccatataa
     1261 atcagcatcc atgttggaat ttaatcgcgg cctagagcaa gacgtttccc gttgaatatg
     1321 gctcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     1381 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gaaaggccag
     1441 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     1501 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     1561 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     1621 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     1681 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     1741 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     1801 cctgtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     1861 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     1921 aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     1981 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gttttttttg tttgcaagca
     2041 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     2101 tgacgctcag tggaacgaaa actcacgtta agggattttg gggatcctcg ggagttgtca
     2161 gcctgtcccg cttataagat catacgccgt tatacgttgt ttacgctttg aggaatccca
     2221 tatggcagaa cattttacag atgcacagat tcagcgctta cgcgagtatg gtaatgggac
     2281 atataaggat atggagttcg cagacgtgag tgcgcgtgaa aaggcgttca cgaagcttat
     2341 gtctgatgcc agtcgcgata atgaaagtgc ccttaaggga atgattgcgc accctgcacg
     2401 ccagggttta agtcgcttaa tgaatgacat tgcagacgct ttggtggctg atggatttat
     2461 tgaagttcgc acacccatta ttatcagtaa agatgcttta gccaaaatga caatcactcc
     2521 cgataaacca ttattcaaac aagtattttg gattgatgat aaacgcgctc tgcgtccaat
     2581 gttagccccg tctctgggca cggtcctgcg tagcctgcgt gatcatactg acggccctgt
     2641 caaaattttc gaaatgggct cttgttttcg caaagaaagc cacagtggga tgcacctgga
     2701 agagtttacc atgttaaacc tggtagatat gggccccgcc ggcgatgcca ctgagtccct
     2761 gaagaaatat attgggatcg taatgaaggc cgccgggctg cctgactacc aattagtcca
     2821 cgaggaatct gatgtgtata aagaaacgat tgatgttgaa atcaacggac aagaggtttg
     2881 tagcgctgct gtaggtcctc actatttgga tgccgcccat gacgtgcacg agccgtgggc
     2941 aggggcaggg ttcggcctgg agcgcctgct gacaattcgc cagggatata gcacagtgat
     3001 gaaaggggga gcttccacaa cctatttgaa cggggctaag atggactaac ctattgaagg
     3061 gttgctaaga ataacaagat caaataagga ggttttttat ggacattacg gaaaaactgc
     3121 gcttgattac gcgcaacgcg gaggaagtcg taacagaaga agagttgcgt cagttgattg
     3181 aaacaaaaga aaaaccccgt gcaattgtag gctatgaacc aagtggcgag atccatttag
     3241 gtcatatgat gaccgtccag aaactgatgg atttacaaga ggcgggattt gagattattg
     3301 tcatgcttgc ggacatcctg gcatacttga atgaaaaagg tacattcgag gagattgccg
     3361 aagtggcaga ctacaataaa aaagtcttca ttgcattagg acttgacgaa agccgcgcca
     3421 aattcgtctt gggtagcgaa tatgagctga gccgtgacta cgtgttagac gtgttaaaga
     3481 tggcccgtat cactacgctt aatcgtgcgc gtcgcagcat ggatgaggtc agccgccgca
     3541 aggaagatcc aatggtttcc caaatgattt acccgcttat gcaggcactg acgggggctc
     3601 atttaggagt ggaccttgcc gtaggtggaa tcgaccaacg taagatccat atgctggcgc
     3661 gcgaaaacct gcctcgcttg ggatatagta gcccggtgtg tttacatacg ccgattctgg
     3721 tggggttaga cggacaaaag atgagtagca gtaagggaaa ttacattagt gttcgtgatc
     3781 ctccagaaga ggtggagcgt aaaatccgta aagcttattg cccggcagga gtggtcgagg
     3841 agaatccgat tctggatatt gccaagtatc acattttgcc ccgttttggt aagatcgtag
     3901 tagagcgcga tgctaaggtt ggaggggacg ttgaatacgc gagtttcgag gaattggcag
     3961 aagactttaa gtctggtcag ctgcatccag ggaggttaaa gatcgcggtc gctaaatatt
     4021 tgaatatgct gttggaggat gcccgtaaac gtctgggcgt gtctgtttaa gtcttccact
     4081 gctagcatta acattagcga ccggcaagca ggatagtaag gaggtttttt atgacggtga
     4141 aatacacaga cgcgcagatt cagcgtttac gcgagtatct cctatctgaa tggggccaaa
     4201 attaattaag gaggacatcc tgccgtttca aacgctaaat tgcctgatgc gctacgctta
     4261 tcaggcctac atgatctctg caatatattg agtttgcgtg cttttgtagg ccggataagg
     4321 cgttcacgcc gcatccggca agaaacagca aacaatccaa aacgccgcgt tcggcggtcg
     4381 acacagatgt aggtgttcca cagggtagcc agcagcatcc tgcgatgcag atccggaaca
     4441 taatggtgca gggcgcttgt ttcggcgtgg gtatggtggc aggccccgtg gccgggggac
     4501 tgttgggcgc tgccggcacc tgtcctacga gttgcatgat aaagaagaca gtcataagtg
     4561 cggcgacgat agtcatgccc cgcgcccacc ggaaggagct accggcagcg gtgcggactg
     4621 ttgtaactca gaataagaaa tgaggccgct catggcgttc tgttgcccgt ctcactggtg
     4681 aaaagaaaaa caaccctggc gccgcttctt tgagcgaacg atcaaaaata agtggcgccc
     4741 catcaaaaaa atattctcaa cataaaaaac tttgtgtaat acttgtaacg ctagatctgg
     4801 gggacggtcc ggcgaccagc gggtctcgaa aacctagcct tgcggggttc gacaccccgg
     4861 tctctcgcca aattttgcac ccagcaaact tggtacgtaa acgcatcgtc ccgccctagc
     4921 tcagaggtag agcgtgcttc tctaaaaagc atggtccccg gttcaaatcc tgggggcggg
     4981 accaactagt atccttagcg aaagctaagg atttttttta agcttggcac tggccgtcgt
     5041 tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca
     5101 tccccctttc gccagacgct ctcccttatg cgactcctgc attaggaagc agcccagtag
     5161 taggttgagg ccgttgagca ccgccgccgc aag
//
