LOCUS       sequence-273020-        2775 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  phage cloning.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2775)
  AUTHORS   Richter MF, Zhao KT, Eton E, Lapinaite A, Newby GA, Thuronyi BW, 
            Wilson C, Koblan LW, Zeng J, Bauer DE, Doudna JA, Liu DR
  TITLE     Phage-assisted evolution of an adenine base editor with improved Cas
            domain compatibility and activity.
  JOURNAL   Nat Biotechnol. 2020 Jul;38(7):883-891. doi: 
            10.1038/s41587-020-0453-z. Epub 2020 Mar 16.
  PUBMED    32433547
REFERENCE   2  (bases 1 to 2775)
  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"; doi: 
            "10.1038/s41587-020-0453-z"; journalName: "Nat Biotechnol"; date: 
            "2020-07"; volume: "38"; issue: "7"; pages: "883-891"
COMMENT     Sequence Label: pBT100.164
FEATURES             Location/Qualifiers
     source          1..2775
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     227..244
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             273..770
                     /codon_start=1
                     /product="DNA adenosine deaminase evolved from the E. coli 
                     tRNA adenosine deaminase TadA (Gaudelli et al., 2017)"
                     /label=TadA*(7.10)
                     /translation="SEVEFSHEYWMRHALTLAKRARDEREVPVGAVLVLNNRVIGEGWN
                     RAIGLHDPTAHAEIMALRQGGLVMQNYRLIDATLYVTFEPCVMCAGAMIHSRIGRVVFG
                     VRNAKTGAAGSLMDVLHYPGMNHRVEITEGILADECAALLCYFFRMPRQVFNAQKKAQS
                     STD"
     CDS             complement(1323..2138)
                     /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(1323..2138)
                     /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     2046..2065
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2130..2149
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      join(2260..2775,1..73)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2749..2768
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
       61 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
      121 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
      181 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
      241 cgagaatgct cttctagtaa ggaggaaaaa tgtcagaagt agagttttca cacgagtact
      301 ggatgagaca cgcattgact ctcgcaaagc gtgctcgaga tgaacgcgag gtgcccgtgg
      361 gagcagtact cgtgctcaac aatcgcgtaa tcggcgaagg ttggaatcgt gcaatcggac
      421 tccacgaccc cactgcacat gcggaaatca tggcccttcg acagggcggg cttgtgatgc
      481 agaattatcg acttatcgat gcgacgctgt acgtcacgtt tgaaccttgc gtaatgtgcg
      541 cgggagctat gattcactcc cgcattggac gagttgtatt cggtgttcgc aacgccaaga
      601 cgggtgccgc aggttcactg atggacgtgc tgcattaccc aggcatgaac caccgggtag
      661 aaatcacaga aggcatattg gcggacgaat gtgcggcgct gttgtgttac ttttttcgca
      721 tgcccaggca ggtctttaac gcccagaaaa aagcacaatc ctctactgac tctggtggtt
      781 cttctggtgg ttcttccggt agcgaaacac cggggacttc agaatcggcc accccggagt
      841 gcctgtcata cgaaaccgag atactgacag tagaatatgg ccttctgcca atcgggaaga
      901 ttgtggagaa acggatagaa tgcacagttt actctgtcga taacaatggt aacatttata
      961 ctcagccagt tgcccagtgg cacgaccggg gagagcagga agtattcgaa tactgtctgg
     1021 aggatggaag tctcattagg gccactaagg accacaaatt tatgacagtc gatggccaga
     1081 tgctgcctat agacgaaatc tttgagcgag agttggacct catgcgagtt gacaaccttc
     1141 ctaattaatg gctgaagagc ctggttaaaa aatgagctga tttaacaaaa atttaacgcg
     1201 aattttaaca aaatattaac gcttacaatt taggtggcac ttttcgggga aatgtgcgcg
     1261 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgaattaat
     1321 tcttagaaaa actcatcgag catcaaatga aactgcaatt tattcatatc aggattatca
     1381 ataccatatt tttgaaaaag ccgtttctgt aatgaaggag aaaactcacc gaggcagttc
     1441 cataggatgg caagatcctg gtatcggtct gcgattccga ctcgtccaac atcaatacaa
     1501 cctattaatt tcccctcgtc aaaaataagg ttatcaagtg agaaatcacc atgagtgacg
     1561 actgaatccg gtgagaatgg caaaagttta tgcatttctt tccagacttg ttcaacaggc
     1621 cagccattac gctcgtcatc aaaatcactc gcatcaacca aaccgttatt cattcgtgat
     1681 tgcgcctgag ccagacgaaa tacgcgatcg ctgttaaaag gacaattaca aacaggaatc
     1741 gaatgcaacc ggcgcaggaa cactgccagc gcatcaacaa tattttcacc tgaatcagga
     1801 tattcttcta atacctggaa tgctgttttc ccggggatcg cagtggtgag taaccatgca
     1861 tcatcaggag tacggataaa atgcttgatg gtcggaagag gcataaattc cgtcagccag
     1921 tttagtctga ccatctcatc tgtaacatca ttggcaacgc tacctttgcc atgtttcaga
     1981 aacaactctg gcgcatcggg cttcccatac aatcgataga ttgtcgcacc tgattgcccg
     2041 acattatcgc gagcccattt atacccatat aaatcagcat ccatgttgga atttaatcgc
     2101 ggcctagagc aagacgtttc ccgttgaata tggctcataa caccccttgt attactgttt
     2161 atgtaagcag acagttttat tgttcatgac caaaatccct taacgtgagt tttcgttcca
     2221 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     2281 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     2341 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     2401 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     2461 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     2521 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     2581 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     2641 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     2701 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     2761 gtatctttat agtcc
//
