LOCUS       sequence-445701-        6133 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  P2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6133)
  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 6133)
  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: pBT128c
FEATURES             Location/Qualifiers
     source          1..6133
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(4516..5331)
                     /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(4516..5331)
                     /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     5239..5258
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5323..5342
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      complement(5472..6017)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(5552..5571)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cacagctaac accacgtcgt ccctatctgc tgccctaggt ctatgagtgg ttgctggata
       61 actttacggg catgcataag gctcgtatga tatattcagg gagaccacaa cggtttccct
      121 ctacaaataa ttttgtttaa cttttactag agtgggaccc tacctgcagg tgcagtaagg
      181 aggaaaaaaa aatgatcaag atagctacaa ggaagtatct tggcaaacaa aacgtttatg
      241 atattggagt cgaaagagat cacaactttg ctctgaagaa cggattcata gcttctaatt
      301 gttttaacaa atactcaata ggcttagcta tcggcacaaa tagcgtcgga tgggcggtga
      361 tcactgatga atataaggtt ccgtctaaaa agttcaaggt tctgggaaat acagaccgcc
      421 acagtatcaa aaaaaatctt ataggggctc ttttatttga cagtggagag acagcggaag
      481 cgactcgtct caaacggaca gctcgtagaa ggtatacacg tcggaagaat cgtatttgtt
      541 atctacagga gattttttca aatgagatgg cgaaagtaga tgatagtttc tttcatcgac
      601 ttgaagagtc ttttttggtg gaagaagaca agaagcatga acgtcatcct atttttggaa
      661 atatagtaga tgaagttgct tatcatgaga aatatccaac tatctatcat ctgcgaaaaa
      721 aattggtaga ttctactgat aaagcggatt tgcgcttaat ctatttggcc ttagcgcata
      781 tgattaagtt tcgtggtcat tttttgattg agggagattt aaatcctgat aatagtgatg
      841 tggacaaact atttatccag ttggtacaaa cctacaatca attatttgaa gaaaacccta
      901 ttaacgcaag tggagtagat gctaaagcga ttctttctgc acgattgagt aaatcaagac
      961 gattagaaaa tctcattgct cagctccccg gtgagaagaa aaatggctta tttgggaatc
     1021 tcattgcttt gtcattgggt ttgaccccta attttaaatc aaattttgat ttggcagaag
     1081 atgctaaatt acagctttca aaagatactt acgatgatga tttagataat ttattggcgc
     1141 aaattggaga tcaatatgct gatttgtttt tggcagctaa gaatttatca gatgctattt
     1201 tactttcaga tatcctaaga gtaaatactg aaataactaa ggctccccta tcagcttcaa
     1261 tgattaaacg ctacgatgaa catcatcaag acttgactct tttaaaagct ttagttcgac
     1321 aacaacttcc agaaaagtat aaagaaatct tttttgatca atcaaaaaac ggatatgcag
     1381 gttatattga tgggggagct agccaagaag aattttataa atttatcaaa ccaattttag
     1441 aaaaaatgga tggtactgag gaattattgg tgaaactaaa tcgtgaagat ttgctgcgca
     1501 agcaacggac ctttgacaac ggctctattc cccatcaaat tcacttgggt gagctgcatg
     1561 ctattttgag aagacaagaa gacttttatc catttttaaa agacaatcgt gagaagattg
     1621 aaaaaatctt gacttttcga attccttatt atgttggtcc attggcgcgt ggcaatagtc
     1681 gttttgcatg gatgactcgg aagtctgaag aaacaattac cccatggaat tttgaagaag
     1741 ttgtcgataa aggtgcttca gctcaatcat ttattgaacg catgacaaac tttgataaaa
     1801 atcttccaaa tgaaaaagta ctaccaaaac atagtttgct ttatgagtat tttacggttt
     1861 ataacgaatt gacaaaggtc aaatatgtta ctgaaggaat gcgaaaacca gcatttcttt
     1921 caggtgaaca gaagaaagcc attgttgatt tactcttcaa aacaaatcga aaagtaaccg
     1981 ttaagcaatt aaaagaagat tatttcaaaa aaatagaatg ttttgatagt gttgaaattt
     2041 caggagttga agatagattt aatgcttcat taggtaccta ccatgatttg ctaaaaatta
     2101 ttaaagataa agattttttg gataatgaag aaaatgaaga tatcttagag gatattgttt
     2161 taacattgac cttatttgaa gatagggaga tgattgagga aagacttaaa acatatgctc
     2221 acctctttga tgataaggtg atgaaacagc ttaaacgtcg ccgttatact ggttggggac
     2281 gtttgtctcg aaaattgatt aatggtatta gggataagca atctggcaaa acaatattag
     2341 attttttgaa atcagatggt tttgccaatc gcaattttat gcagctgatc catgatgata
     2401 gtttgacatt taaagaagac attcaaaaag cacaagtgtc tggacaaggc gatagtttac
     2461 atgaacatat tgcaaattta gctggtagcc ctgctattaa aaaaggtatt ttacagactg
     2521 taaaagttgt tgatgaattg gtcaaagtaa tggggcggca taagccagaa aatatcgtta
     2581 ttgaaatggc acgtgaaaat cagacaactc aaaagggcca gaaaaattcg cgagagcgta
     2641 tgaaacgaat cgaagaaggt atcaaagaat taggaagtca gattcttaaa gagcatcctg
     2701 ttgaaaatac tcaattgcaa aatgaaaagc tctatctcta ttatctccaa aatggaagag
     2761 acatgtatgt ggaccaagaa ttagatatta atcgtttaag tgattatgat gtcgatgcca
     2821 ttgttccaca aagtttcctt aaagacgatt caatagacaa taaggtctta acgcgttctg
     2881 ataaaaatcg tggtaaatcg gataacgttc caagtgaaga agtagtcaaa aagatgaaaa
     2941 actattggag acaacttcta aacgccaagt taatcactca acgtaagttt gataatttaa
     3001 cgaaagctga acgtggaggt ttgagtgaac ttgataaagc tggttttatc aaacgccaat
     3061 tggttgaaac tcgccaaatc actaagcatg tggcacaaat tttggatagt cgcatgaata
     3121 ctaaatacga tgaaaatgat aaacttattc gagaggttaa agtgattacc ttaaaatcta
     3181 aattagtttc tgacttccga aaagatttcc aattctataa agtacgtgag attaacaatt
     3241 accatcatgc ccatgatgcg tatctaaatg ccgtcgttgg aactgctttg attaagaaat
     3301 atccaaaact tgaatcggag tttgtctatg gtgattataa agtttatgat gttcgtaaaa
     3361 tgattgctaa gtctgagcaa gaaataggca aagcaaccgc aaaatatttc ttttactcta
     3421 atatcatgaa cttcttcaaa acagaaatta cacttgcaaa tggagagatt cgcaaacgcc
     3481 ctctaatcga aactaatggg gaaactggag aaattgtctg ggataaaggg cgagattttg
     3541 ccacagtgcg caaagtattg tccatgcccc aagtcaatat tgtcaagaaa acagaagtac
     3601 agacaggcgg attctccaag gagtcaattt taccaaaaag aaattcggac aagcttattg
     3661 ctcgtaaaaa agactgggat ccaaaaaaat atggtggttt tgatagtcca acggtagctt
     3721 attcagtcct agtggttgct aaggtggaaa aagggaaatc gaagaagtta aaatccgtta
     3781 aagagttact agggatcaca attatggaaa gaagttcctt tgaaaaaaat ccgattgact
     3841 ttttagaagc taaaggatat aaggaagtta aaaaagactt aatcattaaa ctacctaaat
     3901 atagtctttt tgagttagaa aacggtcgta aacggatgct ggctagtgcc ggagaattac
     3961 aaaaaggaaa tgagctggct ctgccaagca aatatgtgaa ttttttatat ttagctagtc
     4021 attatgaaaa gttgaagggt agtccagaag ataacgaaca aaaacaattg tttgtggagc
     4081 agcataagca ttatttagat gagattattg agcaaatcag tgaattttct aagcgtgtta
     4141 ttttagcaga tgccaattta gataaagttc ttagtgcata taacaaacat agagacaaac
     4201 caatacgtga acaagcagaa aatattattc atttatttac gttgacgaat cttggagctc
     4261 ccgctgcttt taaatatttt gatacaacaa ttgatcgtaa acgatatacg tctacaaaag
     4321 aagttttaga tgccactctt atccatcaat ccatcactgg tctttatgaa acacgcattg
     4381 atttgagtca gctaggaggt gactaaccag gatagcaacg gtgaaaataa gattaaaatg
     4441 ttataaggca ctcctcagca aatataatga ccctcttgat aacccaagag ggcatttttt
     4501 aatgcccatg gcgtcttaga aaaactcatc gagcatcaaa tgaaactgca atttattcat
     4561 atcaggatta tcaataccat atttttgaaa aagccgtttc tgtaatgaag gagaaaactc
     4621 accgaggcag ttccatagga tggcaagatc ctggtatcgg tctgcgattc cgactcgtcc
     4681 aacatcaata caacctatta atttcccctc gtcaaaaata aggttatcaa gtgagaaatc
     4741 accatgagtg acgactgaat ccggtgagaa tggcaaaagt ttatgcattt ctttccagac
     4801 ttgttcaaca ggccagccat tacgctcgtc atcaaaatca ctcgcatcaa ccaaaccgtt
     4861 attcattcgt gattgcgcct gagcgagacg aaatacgcga tcgctgttaa aaggacaatt
     4921 acaaacagga atcgaatgca accggcgcag gaacactgcc agcgcatcaa caatattttc
     4981 acctgaatca ggatattctt ctaatacctg gaatgctgtt ttcccgggga tcgcagtggt
     5041 gagtaaccat gcatcatcag gagtacggat aaaatgcttg atggtcggaa gaggcataaa
     5101 ttccgtcagc cagtttagtc tgaccatctc atctgtaaca tcattggcaa cgctaccttt
     5161 gccatgtttc agaaacaact ctggcgcatc gggcttccca tacaatcgat agattgtcgc
     5221 acctgattgc ccgacattat cgcgagccca tttataccca tataaatcag catccatgtt
     5281 ggaatttaat cgcggcctag agcaagacgt ttcccgttga atatggctca taacacccct
     5341 tgtattactg tttatgtaag cagacagttt tattgttcat gaagagaaac ctacggatcg
     5401 gggcggagat ttcctggaag atgccaggaa gatacttaac agggaagtga gagggccgcg
     5461 gcaaagccgt ttttccatag gctccgcccc cctgacaagc atcacgaaat ctgacgctca
     5521 aatcagtggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggcggc
     5581 tccctcgtgc gctctcctgt tcctgccttt cggtttaccg gtgtcattcc gctgttatgg
     5641 ccgcgtttgt ctcattccac gcctgacact cagttccggg taggcagttc gctccaagct
     5701 ggactgtatg cacgaacccc ccgttcagtc cgaccgctgc gccttatccg gtaactatcg
     5761 tcttgagtcc aacccggaaa gacatgcaaa agcaccactg gcagcagcca ctggtaattg
     5821 atttagagga gttagtcttg aagtcatgcg ccggttaagg ctaaactgaa aggacaagtt
     5881 ttggtgactg cgctcctcca agccagttac ctcggttcaa agagttggta gctcagagaa
     5941 ccttcgaaaa accgccctgc aaggcggttt tttcgttttc agagcaagag attacgcgca
     6001 gaccaaaacg atctcaagaa gatcatctta ttaaggagcc ccatacgata taagaggaca
     6061 tccggtcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt tgctgaggag
     6121 acttagggac tct
//
