LOCUS       sequence-259782-        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"
     primer_bind     287..306
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     371..390
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      513..1101
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1002..1021
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1255..1272
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(1336..1353)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1506..1592
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1684..1711
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     CDS             2641..2658
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      2688..2719
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     CDS             complement(join(2735..3171,1..379))
                     /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(join(2735..3171,1..379))
                     /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"
ORIGIN
        1 acactgccag cgcatcaaca atattttcac ctgaatcagg atattcttct aatacctgga
       61 atgctgtttt cccggggatc gcagtggtga gtaaccatgc atcatcagga gtacggataa
      121 aatgcttgat ggtcggaaga ggcataaatt ccgtcagcca gtttagtctg accatctcat
      181 ctgtaacatc attggcaacg ctacctttgc catgtttcag aaacaactct ggcgcatcgg
      241 gcttcccata caatcgatag attgtcgcac ctgattgccc gacattatcg cgagcccatt
      301 tatacccata taaatcagca tccatgttgg aatttaatcg cggcctcgag caagacgttt
      361 cccgttgaat atggctcata acaccccttg tattactgtt tatgtaagca gacagtttta
      421 ttgttcatga tgatatattt ttatcttgtg caatgtaaca tcagagattt tgaaggccaa
      481 ataggccgtg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
      541 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
      601 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
      661 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
      721 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
      781 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
      841 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
      901 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
      961 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     1021 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     1081 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     1141 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     1201 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     1261 tcagtgagcg aggaagcgga agagcgccaa gaggacatcc ggtggctgtt ttggcggatg
     1321 agagaagatt ttcagcctga tacagattaa atcagaacgc agaagcggtc tgataaaaca
     1381 gaatttgcct ggcggcagta gcgcggtggt cccacctgac cccatgccga actcagaagt
     1441 gaaacgccgt agcgccgatg gtagtgtggg gtctccccat gcgagagtag ggaactgcca
     1501 ggcatcaaat aaaacgaaag gctcagtcga aagactgggc ctttcgtttt atctgttgtt
     1561 tgtcggtgaa cgctctcctg agtaggacaa atccgccggg agcggatttg aacgttgcga
     1621 agcaacggcc cggagggtgg cgggcaggac gcccgccata aactgccagg catcaaatta
     1681 agcagaaggc catcctgacg gatggccttt ttgcgtttct acaaactcgc tgaggattct
     1741 agagcacagc taacaccacg tcgtccctat ctgctgccct aggtctatga gtggttgctg
     1801 gataacttta cgggcatgca taaggctcgt ataatatatt cagggagacc acaacggttt
     1861 ccctctacaa ataattttgt ttaactttat atctgttatt ttttaaggag gaaaaaaaaa
     1921 tgagcaaagg tgaagagctg tttacgggtg tagtaccgat cttagtggaa ttagacggcg
     1981 acgtgaacgg tcacaaattt agcgtgcgcg gcgaaggcga aggtgacgct accaatggta
     2041 aattgaccct gaagtttatt tgcacaacag gcaaattacc cgttccgtgg cccaccttag
     2101 tgaccaccct gacctatggc gttcagtgct tcagtcgtta cccagatcat atgaaacaac
     2161 acgatttttt caaatcagcc atgcctgaag gatatgttca agagcgtaca atcagcttca
     2221 aggacgatgg cacctataaa acgcgtgcgg aagtgaaatt tgaaggcgac acattagtaa
     2281 accgtatcga actgaaaggt atcgacttca aagaagacgg caacatttta ggccataagc
     2341 tggaatataa ctttaattct cataacgtgt agaattacgg ccgataaaca gaaaaacggt
     2401 atcaaggcaa atttcaaaat tcgccataac gtggaagacg gcagcgttca attagcggat
     2461 cattatcaac aaaacacgcc gattggtgac gggcctgtac tgttacctga caaccactac
     2521 ctgagcaccc agtcagtcct gagcaaagat ccgaacgaaa aacgcgatca catggttctg
     2581 ttagaattcg tgaccgctgc aggcattact cacggaatgg acgaactcta caagggaagc
     2641 catcaccacc accatcactg aacttaatta acggcactcc tcagccaagt caaaagcctc
     2701 cgaccggagg cttttgacta catgcccatg gcgtttagaa aaactcatcg agcatcaaat
     2761 gaaactgcaa tttattcata tcaggattat caataccata tttttgaaaa agccgtttct
     2821 gtaatgaagg agaaaactca ccgaggcagt tccataggat ggcaagatcc tggtatcggt
     2881 ctgcgattcc gactcgtcca acatcaatac aacctattaa tttcccctcg tcaaaaataa
     2941 ggttatcaag tgagaaatca ccatgagtga cgactgaatc cggtgagaat ggcaaaagct
     3001 tatgcatttc tttccagact tgttcaacag gccagccatt acgctcgtca tcaaaatcac
     3061 tcgcatcaac caaaccgtta ttcattcgtg attgcgcctg agcgagacga aatacgcgat
     3121 cgctgttaaa aggacaatta caaacaggaa tcgaatgcaa ccggcgcagg a
//
