LOCUS       sequence-238844-        2958 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  clone APOBEC1 phage by SapI Golden Gate.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2958)
  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 2958)
  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: pBT100.106b
FEATURES             Location/Qualifiers
     source          1..2958
                     /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             45..731
                     /codon_start=1
                     /product="cytidine deaminase (C to U editing enzyme) from 
                     rat"
                     /label=APOBEC-1
                     /note="can use ssDNA as a substrate (Komor et al., 2016)"
                     /translation="MSSETGPVAVDPTLRRRIEPHEFEVFFDPRELRKETCLLYEINWG
                     GRHSIWRHTSQNTNKHVEVNFIEKFTTERYFCPNTRCSITWFLSWSPCGECSRAITEFL
                     SRYPHVTLFIYIARLYHHADPRNRQGLRDLISSGVTIQIMTEQESGYCWRNFVNYSPSN
                     EAHWPRYPHLWVRLYVLELYCIILGLPPCLNILRRKQPQLTFFTIALQSCHYQRLPPHI
                     LWATGLK"
     CDS             complement(1262..2077)
                     /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(1262..2077)
                     /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     1985..2004
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2069..2088
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2199..2787
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2688..2707
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2941..2958
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 aatgctcttc tagtgtacac actaccacgt taaggaggta aagtatgtct tctgaaaccg
       61 gtccggttgc ggttgacccg accctgcgtc gtcgtatcga accgcacgaa ttcgaagttt
      121 tcttcgaccc gcgtgaactg cgtaaagaaa cctgcctgct gtacgaaatc aactggggtg
      181 gtcgtcactc tatctggcgt cacacctctc agaacaccaa caaacacgtt gaagttaact
      241 tcatcgaaaa attcaccacc gaacgttact tctgcccgaa cacccgttgc tctatcacct
      301 ggttcctgtc ttggtcaccg tgcggagaat gctctcgtgc gatcaccgaa ttcctgtctc
      361 gttacccgca cgttaccctg ttcatctaca tcgcgcgtct gtaccaccac gcggacccgc
      421 gtaaccgtca gggtctgcgt gacctgatct cttctggtgt taccatccag atcatgaccg
      481 aacaggaatc tggttactgc tggcgtaact tcgttaacta ctctccgtct aacgaagcgc
      541 actggccgcg ttacccgcac ctgtgggttc gtctgtacgt tctggaactg tactgcatca
      601 tcttaggctt accaccttgt ttaaatattt tacgccgcaa acagccgcag ctgaccttct
      661 tcaccatcgc gctgcagtct tgccactacc agcgtctgcc gccgcacatc ctgtgggcga
      721 ccggtctgaa atccggtagc gaaacaccgg ggacttcaga atcggccacc ccggagtgcc
      781 tgtcatacga aaccgagata ctgacagtag aatatggcct tctgccaatc gggaagattg
      841 tggagaaacg gatagaatgc acagtttact ctgtcgataa caatggtaac atttatactc
      901 agccagttgc ccagtggcac gaccggggag agcaggaagt attcgaatac tgtctggagg
      961 atggaagtct cattagggcc actaaggacc acaaatttat gacagtcgat ggccagatgc
     1021 tgcctataga cgaaatcttt gagcgagagt tggacctcat gcgagttgac aaccttccta
     1081 attaaggcgg ctgaagagcc tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     1141 attttaacaa aatattaacg cttacaattt aggtggcact tttcggggaa atgtgcgcgg
     1201 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgaattaatt
     1261 cttagaaaaa ctcatcgagc atcaaatgaa actgcaattt attcatatca ggattatcaa
     1321 taccatattt ttgaaaaagc cgtttctgta atgaaggaga aaactcaccg aggcagttcc
     1381 ataggatggc aagatcctgg tatcggtctg cgattccgac tcgtccaaca tcaatacaac
     1441 ctattaattt cccctcgtca aaaataaggt tatcaagtga gaaatcacca tgagtgacga
     1501 ctgaatccgg tgagaatggc aaaagtttat gcatttcttt ccagacttgt tcaacaggcc
     1561 agccattacg ctcgtcatca aaatcactcg catcaaccaa accgttattc attcgtgatt
     1621 gcgcctgagc cagacgaaat acgcgatcgc tgttaaaagg acaattacaa acaggaatcg
     1681 aatgcaaccg gcgcaggaac actgccagcg catcaacaat attttcacct gaatcaggat
     1741 attcttctaa tacctggaat gctgttttcc cggggatcgc agtggtgagt aaccatgcat
     1801 catcaggagt acggataaaa tgcttgatgg tcggaagagg cataaattcc gtcagccagt
     1861 ttagtctgac catctcatct gtaacatcat tggcaacgct acctttgcca tgtttcagaa
     1921 acaactctgg cgcatcgggc ttcccataca atcgatagat tgtcgcacct gattgcccga
     1981 cattatcgcg agcccattta tacccatata aatcagcatc catgttggaa tttaatcgcg
     2041 gcctagagca agacgtttcc cgttgaatat ggctcataac accccttgta ttactgttta
     2101 tgtaagcaga cagttttatt gttcatgacc aaaatccctt aacgtgagtt ttcgttccac
     2161 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     2221 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     2281 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     2341 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     2401 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     2461 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     2521 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     2581 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     2641 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     2701 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     2761 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     2821 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     2881 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     2941 agcgagtcag tgagcgag
//
