LOCUS       sequence-263776-        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"
     terminator      654..685
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     CDS             complement(701..1516)
                     /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(701..1516)
                     /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     1424..1443
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1508..1527
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             complement(1627..2577)
                     /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      2625..2847
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
ORIGIN
        1 ttcgtttgtg aatatcaagg ccaatcgtct gacctgcctc aacctcctgt caatgctggc
       61 ggcggctctg gtggtggttc tggtggcggc tctgagggtg gtggctctga gggtggcggt
      121 tctgagggtg gcggctctga gggaggcggt tccggtggtg gctctggttc cggtgatttt
      181 gattatgaaa agatggcaaa cgctaataag ggggctatga ccgaaaatgc cgatgaaaac
      241 gcgctacagt ctgacgctaa aggcaaactt gattctgtcg ctactgatta cggtgctgct
      301 atcgatggtt tcattggtga cgtttccggc cttgctaatg gtaatggtgc tactggtgat
      361 tttgctggct ctaattccca aatggctcaa gtcggtgacg gtgataattc acctttaatg
      421 aataatttcc gtcaatattt accttccctc cctcaatcgg ttgaatgtcg cccttttgtc
      481 tttggcgctg gtaaacctta cgagttcagt atcgactgcg ataagatcaa cctgttccgc
      541 ggtgtctttg cgtttctttt atatgttgcc acctttatgt atgtattttc tacgtttgct
      601 aacatactgc gtaataagga gtcttaaact taattaacgg cactcctcag ccaagtcaaa
      661 agcctccgac cggaggcttt tgactacatg cccatggcgt ttagaaaaac tcatcgagca
      721 tcaaatgaaa ctgcaattta ttcatatcag gattatcaat accatatttt tgaaaaagcc
      781 gtttctgtaa tgaaggagaa aactcaccga ggcagttcca tagaatggca agatcctggt
      841 atcggtctgc gattccgact cgtccaacat caatacaacc tattaatttc ccctcgtcaa
      901 aaataaggtt atcaagtgag aaatcaccat gagtgacgac tgaatccggt gagaatggca
      961 aaagcttatg catttctttc cagacttgtt caacaggcca gccattacgc tcgtcatcaa
     1021 aatcactcgc atcaaccaaa ccgttattca ttcgtgattg cgcctgagcg agacgaaata
     1081 cgcgatcgct gttaaaagga caattacaaa caggaatcga atgcaaccgg cgcaggaaca
     1141 ctgccagcgc atcaacaata ttttcacctg aatcaggata ttcttctaat acctggaatg
     1201 ctgttttccc ggggatcgca gtggtgagta accatgcatc atcaggagta cggataaaat
     1261 gcttgatggt cggaagaggc ataaattccg tcagccagtt tagtctgacc atctcatctg
     1321 taacatcatt ggcaacgcta cctttgccat gtttcagaaa caactctggc gcatcgggct
     1381 tcccatacaa tcgatagatt gtcgcacctg attgcccgac attatcgcga gcccatttat
     1441 acccatataa atcagcatcc atgttggaat ttaatcgcgg cctcgagcaa gacgtttccc
     1501 gttgaatatg gctcataaca ccccttgtat tactgtttat gtaagcagac agttttattg
     1561 ttcatgatga tatattttta tcttgtgcaa tgtaacatca gagattttga aggccaaata
     1621 ggccgttcag atccttccgt atttagccag tatgttctct agtgtggttc gttgtttttg
     1681 cgtgagccat gagaacgaac cattgagatc atgcttactt tgcatgtcac tcaaaaattt
     1741 tgcctcaaaa ctggtgagct gaatttttgc agttaaagca tcgtgtagtg tttttcttag
     1801 tccgttacgt aggtaggaat ctgatgtaat ggttgttggt attttgtcac cattcatttt
     1861 tatctggttg ttctcaagtt cggttacgag atccatttgt ctatctagtt caacttggaa
     1921 aatcaacgta tcagtcgggc ggcctcgctt atcaaccacc aatttcatat tgctgtaagt
     1981 gtttaaatct ttacttattg gtttcaaaac ccattggtta agccttttaa actcatggta
     2041 gttattttca agcattaaca tgaacttaaa ttcatcaagg ctaatctcta tatttgcctt
     2101 gtgagttttc ttttgtgtta gttcttttaa taaccactca taaatcctca tagagtattt
     2161 gttttcaaaa gacttaacat gttccagatt atattttatg aattttttta actggaaaag
     2221 ataaggcaat atctcttcac taaaaactaa ttctaatttt tcgcttgaga acttggcata
     2281 gtttgtccac tggaaaatct caaagccttt aaccaaagga ttcctgattt ccacagttct
     2341 cgtcatcagc tctctggttg ctttagctaa tacaccataa gcattttccc tactgatgtt
     2401 catcatctga gcgtattggt tataagtgaa cgataccgtc cgttctttcc ttgtagggtt
     2461 ttcaatcgtg gggttgagta gtgccacaca gcataaaatt agcttggttt catgctccgt
     2521 taagtcatag cgactaatcg ctagttcatt tgctttgaaa acaactaatt cagacataca
     2581 tctcaattgg tctaggtgat tttaatcact ataccaattg agatgggcta gtcaatgata
     2641 attactagtc cttttccttt gagttgtggg tatctgtaaa ttctgctaga cctttgctgg
     2701 aaaacttgta aattctgcta gaccctctgt aaattccgct agacctttgt gtgttttttt
     2761 tgtttatatt caagtggtta taatttatag aataaagaaa gaataaaaaa agataaaaag
     2821 aatagatccc agccctgtgt ataactcact actttagtca gttccgcagt attacaaaag
     2881 gatgtcgcaa acgctgtttg ctcctctaca aaacagacct taaaacccta aaggcttaag
     2941 tagcaccctc gcaagctcgg gcaaatcgct gaatattcct tttgtctccg accatcaggc
     3001 acctgagtcg ctgtcttttt cgtgacattc agttcgctgc gctcacggct ctggcagtga
     3061 atgggggtaa atggcactac aggcgccttt tatggattca tgcaaggaaa ctacccataa
     3121 tacaagaaaa gcccgtcacg ggcttctcag ggcgttttat ggcgggtctg ctatgtggtg
     3181 ctatctgact ttttgctgtt cagcagttcc tgccctctga ttttccagtc tgaccacttc
     3241 ggattatccc gtgacaggtc attcagactg gctaatgcac ccagtaaggc agcggtatca
     3301 tcaacaggct tacccgtctt actgtcaaga ggacatccgg tttgttcaga acgctcggtc
     3361 ttgcacaccg ggcgtttttt ctttgtgagt ccatagtaca cttgaataaa taaaaacagc
     3421 cgttgccaga aagaggcacg gctgttttta ttttagactt agggaccctc acagctaaca
     3481 ccacgtcgtc cctatctgct gccctaggtc tatgagtggt tgctggataa ctttacgggc
     3541 atgcataagg ctcgtataat atattcaggg agaccacaac ggtttccctc tacaaataat
     3601 tttgtttaac tttatatctg ttatttttta aggaggaaaa aaaaatgaaa aaattattat
     3661 tcgcaattcc tttagttgtt cctttctatt agacactccg ctgaaactgt tgaaagttgt
     3721 ttagcaaaat agacatacag aaaattagat ttactaacgt ctggaaagac gacaaaactt
     3781 tagatcgtta cgctaactat gagggctgtc tgtggaatgc tacaggcgtt gtagtttgta
     3841 ctggtgacga aactcagtgt tacggtacat gggttcctat tgggcttgct atccctgaaa
     3901 atgagggtgg tggctctgag ggtggtggtt ctgagggtgg cggttctgag ggtggcggta
     3961 ctaaacctcc tgagtacggt gatacaccta ttccgggcta tacttatatc aaccctctcg
     4021 acggcactta tccgcctggt actgagcaaa accccgctaa tcctaatcct tctcttgagg
     4081 agtctcagcc tcttaatact ttcatgtttc agaataatag gttccgaaat aggcaggggg
     4141 cattaactgt ttatacgggc actgttactc aaggcactga ccccgttaaa acttattacc
     4201 agtacactcc tgtatcatca aaagccatgt atgacgctta ctggaacggt aaattcagag
     4261 actgcgcttt ccattctggc tttaatgagg atcca
//
