LOCUS       sequence-269555-        3171 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  sfGFP expression plasmid for characterizing qtRNA charging.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3171)
  AUTHORS   DeBenedictis EA, Carver GD, Chung CZ, Soll D, Badran AH
  TITLE     Multiplex suppression of four quadruplet codons via tRNA directed 
            evolution.
  JOURNAL   Nat Commun. 2021 Sep 29;12(1):5706. doi: 10.1038/s41467-021-25948-y.
  PUBMED    34588441
REFERENCE   2  (bases 1 to 3171)
  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"; journalName: "Nat 
            Commun."; date: "2021-09-29"; volume: "12(1):5706. doi"; pages: " 
            10.1038/s41467-021-25948-y"
COMMENT     Sequence Label: pED17x11
FEATURES             Location/Qualifiers
     source          1..3171
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(127..154)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(246..332)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     485..502
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     primer_bind     complement(566..583)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(737..1325)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(817..836)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(1448..1467)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1459..2274
                     /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             1459..2274
                     /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     complement(1532..1551)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      complement(2290..2321)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     CDS             complement(2351..2368)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
ORIGIN
        1 atattatacg agccttatgc atgcccgtaa agttatccag caaccactca tagacctagg
       61 gcagcagata gggacgacgt ggtgttagct gtgctctaga atcctcagcg agtttgtaga
      121 aacgcaaaaa ggccatccgt caggatggcc ttctgcttaa tttgatgcct ggcagtttat
      181 ggcgggcgtc ctgcccgcca ccctccgggc cgttgcttcg caacgttcaa atccgctccc
      241 ggcggatttg tcctactcag gagagcgttc accgacaaac aacagataaa acgaaaggcc
      301 cagtctttcg actgagcctt tcgttttatt tgatgcctgg cagttcccta ctctcgcatg
      361 gggagacccc acactaccat cggcgctacg gcgtttcact tctgagttcg gcatggggtc
      421 aggtgggacc accgcgctac tgccgccagg caaattctgt tttatcagac cgcttctgcg
      481 ttctgattta atctgtatca ggctgaaaat cttctctcat ccgccaaaac agccaccgga
      541 tgtcctcttg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
      601 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
      661 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
      721 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
      781 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
      841 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
      901 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
      961 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     1021 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     1081 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     1141 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     1201 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     1261 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     1321 ctcaagaaga tcctttgatc ttttctacac ggcctatttg gccttcaaaa tctctgatgt
     1381 tacattgcac aagataaaaa tatatcatca tgaacaataa aactgtctgc ttacataaac
     1441 agtaatacaa ggggtgttat gagccatatt caacgggaaa cgtcttgctc gaggccgcga
     1501 ttaaattcca acatggatgc tgatttatat gggtataaat gggctcgcga taatgtcggg
     1561 caatcaggtg cgacaatcta tcgattgtat gggaagcccg atgcgccaga gttgtttctg
     1621 aaacatggca aaggtagcgt tgccaatgat gttacagatg agatggtcag actaaactgg
     1681 ctgacggaat ttatgcctct tccgaccatc aagcatttta tccgtactcc tgatgatgca
     1741 tggttactca ccactgcgat ccccgggaaa acagcattcc aggtattaga agaatatcct
     1801 gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt gcattcgatt
     1861 cctgtttgta attgtccttt taacagcgat cgcgtatttc gtctcgctca ggcgcaatca
     1921 cgaatgaata acggtttggt tgatgcgagt gattttgatg acgagcgtaa tggctggcct
     1981 gttgaacaag tctggaaaga aatgcataag cttttgccat tctcaccgga ttcagtcgtc
     2041 actcatggtg atttctcact tgataacctt atttttgacg aggggaaatt aataggttgt
     2101 attgatgttg gacgagtcgg aatcgcagac cgataccagg atcttgccat tctatggaac
     2161 tgcctcggtg agttttctcc ttcattacag aaacggcttt ttcaaaaata tggtattgat
     2221 aatcctgata tgaataaatt gcagtttcat ttgatgctcg atgagttttt ctaaacgcca
     2281 tgggcatgta gtcaaaagcc tccggtcgga ggcttttgac ttggctgagg agtgccgtta
     2341 attaagttca gtgatggtgg tggtgatggc ttcccttgta gagttcgtcc attccgtgag
     2401 taatgcctgc agcggtcacg aattctaaca gaaccatgtg atcgcgtttt tcgttcggat
     2461 ctttgctcag gactgactgg gtgctcaggt agtggttgtc aggtaacagt acaggcccgt
     2521 caccaatcgg cgtgttttgt tgataatgat ccgctaattg aacgctgccg tcttccacgt
     2581 tatggcgaat tttgaaattt gccttgatac cgtttttctg tttatcggcc gtaattctac
     2641 acgttatgag aattaaagtt atattccagc ttatggccta aaatgttgcc gtcttctttg
     2701 aagtcgatac ctttcagttc gatacggttt actaatgtgt cgccttcaaa tttcacttcc
     2761 gcacgcgttt tataggtgcc atcgtccttg aagctgattg tacgctcttg aacatatcct
     2821 tcaggcatgg ctgatttgaa aaaatcgtgt tgtttcatat gatctgggta acgactgaag
     2881 cactgaacgc cataggtcag ggtggtcact aaggtgggcc acggaacggg taatttgcct
     2941 gttgtgcaaa taaacttcag ggtcaattta ccattggtag cgtcaccttc gccttcgccg
     3001 cgcacgctaa atttgtgacc gttcacgtcg ccgtctaatt ccactaagat cggtactaca
     3061 cccgtaaaca gctcttcacc tttgctcatt tttttttcct ccttaaaaaa taacagatat
     3121 aaagttaaac aaaattattt gtagagggaa accgttgtgg tctccctgaa t
//
