LOCUS       sequence-263774-        4293 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lenient phage propagation reporter containing pIII-29-TAGA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4293)
  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 4293)
  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: pED7x10
FEATURES             Location/Qualifiers
     source          1..4293
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      516..547
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     CDS             complement(563..1378)
                     /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(563..1378)
                     /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     1286..1305
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1370..1389
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             complement(1489..2439)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFKIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLRNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKHDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      2487..2709
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
ORIGIN
        1 gagggaggcg gttccggtgg tggctctggt tccggtgatt ttgattatga aaagatggca
       61 aacgctaata agggggctat gaccgaaaat gccgatgaaa acgcgctaca gtctgacgct
      121 aaaggcaaac ttgattctgt cgctactgat tacggtgctg ctatcgatgg tttcattggt
      181 gacgtttccg gccttgctaa tggtaatggt gctactggtg attttgctgg ctctaattcc
      241 caaatggctc aagtcggtga cggtgataat tcacctttaa tgaataattt ccgtcaatat
      301 ttaccttccc tccctcaatc ggttgaatgt cgcccttttg tctttggcgc tggtaaacct
      361 tacgagttca gtatcgactg cgataagatc aacctgttcc gcggtgtctt tgcgtttctt
      421 ttatatgttg ccacctttat gtatgtattt tctacgtttg ctaacatact gcgtaataag
      481 gagtcttaaa cttaattaac ggcactcctc agccaagtca aaagcctccg accggaggct
      541 tttgactaca tgcccatggc gtttagaaaa actcatcgag catcaaatga aactgcaatt
      601 tattcatatc aggattatca ataccatatt tttgaaaaag ccgtttctgt aatgaaggag
      661 aaaactcacc gaggcagttc catagaatgg caagatcctg gtatcggtct gcgattccga
      721 ctcgtccaac atcaatacaa cctattaatt tcccctcgtc aaaaataagg ttatcaagtg
      781 agaaatcacc atgagtgacg actgaatccg gtgagaatgg caaaagctta tgcatttctt
      841 tccagacttg ttcaacaggc cagccattac gctcgtcatc aaaatcactc gcatcaacca
      901 aaccgttatt cattcgtgat tgcgcctgag cgagacgaaa tacgcgatcg ctgttaaaag
      961 gacaattaca aacaggaatc gaatgcaacc ggcgcaggaa cactgccagc gcatcaacaa
     1021 tattttcacc tgaatcagga tattcttcta atacctggaa tgctgttttc ccggggatcg
     1081 cagtggtgag taaccatgca tcatcaggag tacggataaa atgcttgatg gtcggaagag
     1141 gcataaattc cgtcagccag tttagtctga ccatctcatc tgtaacatca ttggcaacgc
     1201 tacctttgcc atgtttcaga aacaactctg gcgcatcggg cttcccatac aatcgataga
     1261 ttgtcgcacc tgattgcccg acattatcgc gagcccattt atacccatat aaatcagcat
     1321 ccatgttgga atttaatcgc ggcctcgagc aagacgtttc ccgttgaata tggctcataa
     1381 caccccttgt attactgttt atgtaagcag acagttttat tgttcatgat gatatatttt
     1441 tatcttgtgc aatgtaacat cagagatttt gaaggccaaa taggccgttc agatccttcc
     1501 gtatttagcc agtatgttct ctagtgtggt tcgttgtttt tgcgtgagcc atgagaacga
     1561 accattgaga tcatgcttac tttgcatgtc actcaaaaat tttgcctcaa aactggtgag
     1621 ctgaattttt gcagttaaag catcgtgtag tgtttttctt agtccgttac gtaggtagga
     1681 atctgatgta atggttgttg gtattttgtc accattcatt tttatctggt tgttctcaag
     1741 ttcggttacg agatccattt gtctatctag ttcaacttgg aaaatcaacg tatcagtcgg
     1801 gcggcctcgc ttatcaacca ccaatttcat attgctgtaa gtgtttaaat ctttacttat
     1861 tggtttcaaa acccattggt taagcctttt aaactcatgg tagttatttt caagcattaa
     1921 catgaactta aattcatcaa ggctaatctc tatatttgcc ttgtgagttt tcttttgtgt
     1981 tagttctttt aataaccact cataaatcct catagagtat ttgttttcaa aagacttaac
     2041 atgttccaga ttatatttta tgaatttttt taactggaaa agataaggca atatctcttc
     2101 actaaaaact aattctaatt tttcgcttga gaacttggca tagtttgtcc actggaaaat
     2161 cttaaagcct ttaaccaaag gattcctgat ttccacagtt ctcgtcatca gctctctggt
     2221 tgctttagct aatacaccat aagcattttc cctactgatg ttcatcatct gagcgtattg
     2281 gttataagtg aacgataccg tccgttcttt ccttgtaggg ttttcaatcg tggggttgag
     2341 tagtgccaca cagcataaaa ttagcttggt ttcatgctcc gttaagtcat agcgactaat
     2401 cgctagttca tttgctttga aaacaactaa ttcagacata catctcaatt ggtctaggtg
     2461 attttaatca ctataccaat tgagatgggc tagtcaatga taattactag tccttttcct
     2521 ttgagttgtg ggtatctgta aattctgcta gacctttgct ggaaaacttg taaattctgc
     2581 tagaccctct gtaaattccg ctagaccttt gtgtgttttt tttgtttata ttcaagtggt
     2641 tataatttat agaataaaga aagaataaaa aaagataaaa agaatagatc ccagccctgt
     2701 gtataactca ctactttagt cagttccgca gtattacaaa aggatgtcgc aaacgctgtt
     2761 tgctcctcta caaaacagac cttaaaaccc taaaggctta agtagcaccc tcgcaagctc
     2821 gggcaaatcg ctgaatattc cttttgtctc cgaccatcag gcacctgagt cgctgtcttt
     2881 ttcgtgacat tcagttcgct gcgctcacgg ctctggcagt gaatgggggt aaatggcact
     2941 acaggcgcct tttatggatt catgcaagga aactacccat aatacaagaa aagcccgtca
     3001 cgggcttctc agggcgtttt atggcgggtc tgctatgtgg tgctatctga ctttttgctg
     3061 ttcagcagtt cctgccctct gattttccag tctgaccact tcggattatc ccgtgacagg
     3121 tcattcagac tggctaatgc acccagtaag gcagcggtat catcaacagg cttacccgtc
     3181 ttactgtcaa gaggacatcc ggtttgttca gaacgctcgg tcttgcacac cgggcgtttt
     3241 ttctttgtga gtccatagta cacttgaata aataaaaaca gccgttgcca gaaagaggca
     3301 cggctgtttt tattttagac ttagggaccc tcacagctaa caccacgtcg tccctatctg
     3361 ctgccctagg tctatgagtg gttgctggat aactttacgg gcatgcataa ggctcgtata
     3421 atatattcag ggagaccaca acggtttccc tctacaaata attttgttta actttatatc
     3481 tgttattttt taaggaggaa aaaaaaatga aaaaattatt attcgcaatt cctttagttg
     3541 ttcctttcta ttctcactcc gctgaaactg ttgaaagttg tttagcaaaa tagacataca
     3601 gaaaattcat ttactaacgt ctggaaagac gacaaaactt tagatcgtta cgctaactat
     3661 gagggctgtc tgtggaatgc tacaggcgtt gtagtttgta ctggtgacga aactcagtgt
     3721 tacggtacat gggttcctat tgggcttgct atccctgaaa atgagggtgg tggctctgag
     3781 ggtggtggtt ctgagggtgg cggttctgag ggtggcggta ctaaacctcc tgagtacggt
     3841 gatacaccta ttccgggcta tacttatatc aaccctctcg acggcactta tccgcctggt
     3901 actgagcaaa accccgctaa tcctaatcct tctcttgagg agtctcagcc tcttaatact
     3961 ttcatgtttc agaataatag gttccgaaat aggcaggggg cattaactgt ttatacgggc
     4021 actgttactc aaggcactga ccccgttaaa acttattacc agtacactcc tgtatcatca
     4081 aaagccatgt atgacgctta ctggaacggt aaattcagag actgcgcttt ccattctggc
     4141 tttaatgagg atccattcgt ttgtgaatat caaggccaat cgtctgacct gcctcaacct
     4201 cctgtcaatg ctggcggcgg ctctggtggt ggttctggtg gcggctctga gggtggtggc
     4261 tctgagggtg gcggttctga gggtggcggc tct
//
