LOCUS       sequence-238876-        2760 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  clone FERNY phage by SapI Golden Gate.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2760)
  AUTHORS   Thuronyi BW, Koblan LW, Levy JM, Yeh WH, Zheng C, Newby GA, Wilson 
            C, Bhaumik M, Shubina-Oleinik O, Holt JR, Liu DR
  TITLE     Continuous evolution of base editors with expanded target 
            compatibility and improved activity.
  JOURNAL   Nat Biotechnol. 2019 Jul 22. pii: 10.1038/s41587-019-0193-0. doi: 
            10.1038/s41587-019-0193-0.
  PUBMED    31332326
REFERENCE   2  (bases 1 to 2760)
  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 
            Biotechnol. 2019 Jul 22. pii: 10.1038/s41587-019-0193-0. doi: 
            10.1038/s41587-019-0193-0."
COMMENT     Sequence Label: pBT100.136c
FEATURES             Location/Qualifiers
     source          1..2760
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             31..39
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             complement(1064..1879)
                     /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(1064..1879)
                     /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     1787..1806
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1871..1890
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2001..2589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2490..2509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2743..2760
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 aatgctcttc tagtgtacac actaccacgt taaggaggta aagtatgttt gaacgaaatt
       61 atgacccacg cgagctgcgc aaggaaacat atcttcttta cgagattaag tggggcaaga
      121 gtggtaaact ctggaggcat tggtgtcaaa acaatcgcac tcaacatgca gaggtgtatt
      181 tcctggaaaa catttttaac gctcgccgct ttaatccatc aacgcattgc tccatcacgt
      241 ggtatttgag ttggagtccc tgcgctgaat gtagccaaaa gatcgttgac tttcttaaag
      301 agcaccccaa cgtcaacctt gagatatacg tcgctcgact gtattaccat gaagatgaaa
      361 ggaatagaca ggggctccga gatctcgtga actctggagt cactatacgc attatggatc
      421 tgcccgatta caactattgc tggaaaactt tcgtaagtga ccaaggagga gacgaggatt
      481 actggccagg gcattttgcg ccttggataa aacagtatag cttgaagctg aaatccggta
      541 gtgaaacacc ggggacttca gaatcggcca ccccggagtg cctgtcatac gaaaccgaga
      601 tactgacagt agaatatggc cttctgccaa tcgggaagat tgtggagaaa cggatagaat
      661 gcacagttta ctctgtcgat aacaatggta acatttatac tcagccagtt gcccagtggc
      721 acgaccgggg agagcaggaa gtattcgaat actgtctgga ggatggaagt ctcattaggg
      781 ccactaagga ccacaaattt atgacagtcg atggccagat gctgcctata gacgaaatct
      841 ttgagcgaga gttggacctc atgcgagttg acaaccttcc taattaaggc ggctgaagag
      901 cctggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa
      961 cgcttacaat ttaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     1021 tttctaaata cattcaaata tgtatccgct catgaattaa ttcttagaaa aactcatcga
     1081 gcatcaaatg aaactgcaat ttattcatat caggattatc aataccatat ttttgaaaaa
     1141 gccgtttctg taatgaagga gaaaactcac cgaggcagtt ccataggatg gcaagatcct
     1201 ggtatcggtc tgcgattccg actcgtccaa catcaataca acctattaat ttcccctcgt
     1261 caaaaataag gttatcaagt gagaaatcac catgagtgac gactgaatcc ggtgagaatg
     1321 gcaaaagttt atgcatttct ttccagactt gttcaacagg ccagccatta cgctcgtcat
     1381 caaaatcact cgcatcaacc aaaccgttat tcattcgtga ttgcgcctga gccagacgaa
     1441 atacgcgatc gctgttaaaa ggacaattac aaacaggaat cgaatgcaac cggcgcagga
     1501 acactgccag cgcatcaaca atattttcac ctgaatcagg atattcttct aatacctgga
     1561 atgctgtttt cccggggatc gcagtggtga gtaaccatgc atcatcagga gtacggataa
     1621 aatgcttgat ggtcggaaga ggcataaatt ccgtcagcca gtttagtctg accatctcat
     1681 ctgtaacatc attggcaacg ctacctttgc catgtttcag aaacaactct ggcgcatcgg
     1741 gcttcccata caatcgatag attgtcgcac ctgattgccc gacattatcg cgagcccatt
     1801 tatacccata taaatcagca tccatgttgg aatttaatcg cggcctagag caagacgttt
     1861 cccgttgaat atggctcata acaccccttg tattactgtt tatgtaagca gacagtttta
     1921 ttgttcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     1981 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     2041 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     2101 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     2161 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     2221 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     2281 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     2341 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     2401 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     2461 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     2521 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     2581 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     2641 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     2701 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
//
