LOCUS       sequence-259790-        4295 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Stringent phage propagation reporter containing pIII-16-29-34-TAGA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4295)
  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 4295)
  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: pED7x26
FEATURES             Location/Qualifiers
     source          1..4295
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     152..171
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     236..255
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             complement(355..1305)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLRNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKHDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      1353..1575
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      3677..3708
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     CDS             complement(join(3724..4295,1..244))
                     /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(3724..4295,1..244))
                     /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 gaagaggcat aaattccgtc agccagttta gtctgaccat ctcatctgta acatcattgg
       61 caacgctacc tttgccatgt ttcagaaaca actctggcgc atcgggcttc ccatacaatc
      121 gatagattgt cgcacctgat tgcccgacat tatcgcgagc ccatttatac ccatataaat
      181 cagcatccat gttggaattt aatcgcggcc tcgagcaaga cgtttcccgt tgaatatggc
      241 tcataacacc ccttgtatta ctgtttatgt aagcagacag ttttattgtt catgatgata
      301 tatttttatc ttgtgcaatg taacatcaga gattttgaag gccaaatagg ccgttcagat
      361 ccttccgtat ttagccagta tgttctctag tgtggttcgt tgtttttgcg tgagccatga
      421 gaacgaacca ttgagatcat gcttactttg catgtcactc aaaaattttg cctcaaaact
      481 ggtgagctga atttttgcag ttaaagcatc gtgtagtgtt tttcttagtc cgttacgtag
      541 gtaggaatct gatgtaatgg ttgttggtat tttgtcacca ttcattttta tctggttgtt
      601 ctcaagttcg gttacgagat ccatttgtct atctagttca acttggaaaa tcaacgtatc
      661 agtcgggcgg cctcgcttat caaccaccaa tttcatattg ctgtaagtgt ttaaatcttt
      721 acttattggt ttcaaaaccc attggttaag ccttttaaac tcatggtagt tattttcaag
      781 cattaacatg aacttaaatt catcaaggct aatctctata tttgccttgt gagttttctt
      841 ttgtgttagt tcttttaata accactcata aatcctcata gagtatttgt tttcaaaaga
      901 cttaacatgt tccagattat attttatgaa tttttttaac tggaaaagat aaggcaatat
      961 ctcttcacta aaaactaatt ctaatttttc gcttgagaac ttggcatagt ttgtccactg
     1021 gaaaatctca aagcctttaa ccaaaggatt cctgatttcc acagttctcg tcatcagctc
     1081 tctggttgct ttagctaata caccataagc attttcccta ctgatgttca tcatctgagc
     1141 gtattggtta taagtgaacg ataccgtccg ttctttcctt gtagggtttt caatcgtggg
     1201 gttgagtagt gccacacagc ataaaattag cttggtttca tgctccgtta agtcatagcg
     1261 actaatcgct agttcatttg ctttgaaaac aactaattca gacatacatc tcaattggtc
     1321 taggtgattt taatcactat accaattgag atgggctagt caatgataat tactagtcct
     1381 tttcctttga gttgtgggta tctgtaaatt ctgctagacc tttgctggaa aacttgtaaa
     1441 ttctgctaga ccctctgtaa attccgctag acctttgtgt gttttttttg tttatattca
     1501 agtggttata atttatagaa taaagaaaga ataaaaaaag ataaaaagaa tagatcccag
     1561 ccctgtgtat aactcactac tttagtcagt tccgcagtat tacaaaagga tgtcgcaaac
     1621 gctgtttgct cctctacaaa acagacctta aaaccctaaa ggcttaagta gcaccctcgc
     1681 aagctcgggc aaatcgctga atattccttt tgtctccgac catcaggcac ctgagtcgct
     1741 gtctttttcg tgacattcag ttcgctgcgc tcacggctct ggcagtgaat gggggtaaat
     1801 ggcactacag gcgcctttta tggattcatg caaggaaact acccataata caagaaaagc
     1861 ccgtcacggg cttctcaggg cgttttatgg cgggtctgct atgtggtgct atctgacttt
     1921 ttgctgttca gcagttcctg ccctctgatt ttccagtctg accacttcgg attatcccgt
     1981 gacaggtcat tcagactggc taatgcaccc agtaaggcag cggtatcatc aacaggctta
     2041 cccgtcttac tgtcaagagg acatccggtt tgttcagaac gctcggtctt gcacaccggg
     2101 cgttttttct ttgtgagtcc atagtacact tgaataaata aaaacagccg ttgccagaaa
     2161 gaggcacggc tgtttttatt ttagacttag ggaccctcac agctaacacc acgtcgtccc
     2221 tatctgctgc cctaggtcta tgagtggttg ctggataact ttacgggcat gcataaggct
     2281 cgtataatat attcagggag accacaacgg tttccctcta caaataattt tgtttaactt
     2341 tatatctgtt attttttaag gaggaaaaaa aaatgaaaaa attattattc gcaattcctt
     2401 tagttgttcc tttctattag acactccgct gaaactgttg aaagttgttt agcaaaatag
     2461 acatacagaa aattagattt actaacgtct ggaaagacga caaaacttta gatcgttacg
     2521 ctaactatga gggctgtctg tggaatgcta caggcgttgt agtttgtact ggtgacgaaa
     2581 ctcagtgtta cggtacatgg gttcctattg ggcttgctat ccctgaaaat gagggtggtg
     2641 gctctgaggg tggtggttct gagggtggcg gttctgaggg tggcggtact aaacctcctg
     2701 agtacggtga tacacctatt ccgggctata cttatatcaa ccctctcgac ggcacttatc
     2761 cgcctggtac tgagcaaaac cccgctaatc ctaatccttc tcttgaggag tctcagcctc
     2821 ttaatacttt catgtttcag aataataggt tccgaaatag gcagggggca ttaactgttt
     2881 atacgggcac tgttactcaa ggcactgacc ccgttaaaac ttattaccag tacactcctg
     2941 tatcatcaaa agccatgtat gacgcttact ggaacggtaa attcagagac tgcgctttcc
     3001 attctggctt taatgaggat ccattcgttt gtgaatatca aggccaatcg tctgacctgc
     3061 ctcaacctcc tgtcaatgct ggcggcggct ctggtggtgg ttctggtggc ggctctgagg
     3121 gtggtggctc tgagggtggc ggttctgagg gtggcggctc tgagggaggc ggttccggtg
     3181 gtggctctgg ttccggtgat tttgattatg aaaagatggc aaacgctaat aagggggcta
     3241 tgaccgaaaa tgccgatgaa aacgcgctac agtctgacgc taaaggcaaa cttgattctg
     3301 tcgctactga ttacggtgct gctatcgatg gtttcattgg tgacgtttcc ggccttgcta
     3361 atggtaatgg tgctactggt gattttgctg gctctaattc ccaaatggct caagtcggtg
     3421 acggtgataa ttcaccttta atgaataatt tccgtcaata tttaccttcc ctccctcaat
     3481 cggttgaatg tcgccctttt gtctttggcg ctggtaaacc ttacgagttc agtatcgact
     3541 gcgataagat caacctgttc cgcggtgtct ttgcgtttct tttatatgtt gccaccttta
     3601 tgtatgtatt ttctacgttt gctaacatac tgcgtaataa ggagtcttaa acttaattaa
     3661 cggcactcct cagccaagtc aaaagcctcc gaccggaggc ttttgactac atgcccatgg
     3721 cgtttagaaa aactcatcga gcatcaaatg aaactgcaat ttattcatat caggattatc
     3781 aataccatat ttttgaaaaa gccgtttctg taatgaagga gaaaactcac cgaggcagtt
     3841 ccataggatg gcaagatcct ggtatcggtc tgcgattccg actcgtccaa catcaataca
     3901 acctattaat ttcccctcgt caaaaataag gttatcaagt gagaaatcac catgagtgac
     3961 gactgaatcc ggtgagaatg gcaaaagctt atgcatttct ttccagactt gttcaacagg
     4021 ccagccatta cgctcgtcat caaaatcact cgcatcaacc aaaccgttat tcattcgtga
     4081 ttgcgcctga gcgagacgaa atacgcgatc gctgttaaaa ggacaattac aaacaggaat
     4141 cgaatgcaac cggcgcagga acactgccag cgcatcaaca atattttcac ctgaatcagg
     4201 atattcttct aatacctgga atgctgtttt cccggggatc gcagtggtga gtaaccatgc
     4261 atcatcagga gtacggataa aatgcttgat ggtcg
//
