LOCUS       sequence-243370-         898 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Bacterial expression of A3A with N-terminal MBP an C-terminal His 
            tag, both cleavable with TEV protease.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 898)
  AUTHORS   Schutsky EK, DeNizio JE, Hu P, Liu MY, Nabel CS, Fabyanic EB, Hwang 
            Y, Bushman FD, Wu H, Kohli RM
  TITLE     Nondestructive, base-resolution sequencing of 
            5-hydroxymethylcytosine using a DNA deaminase.
  JOURNAL   Nat Biotechnol. 2018 Oct 8. pii: nbt.4204. doi: 10.1038/nbt.4204.
  PUBMED    30295673
REFERENCE   2  (bases 1 to 898)
  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. 2018 Oct 8. pii: nbt.4204. doi: 10.1038/nbt.4204."
COMMENT     Sequence Label: pET41-MBP-t-A3A-t-His
FEATURES             Location/Qualifiers
     source          1..898
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             33..44
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             57..77
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             84..680
                     /codon_start=1
                     /gene="human APOBEC3A"
                     /gene_synonym="A3A"
                     /product="human cytidine deaminase (C to U editing enzyme) 
                     that efficiently edits methylated cytosines (Wang et al., 
                     2018)"
                     /label=APOBEC3A
                     /note="also known as hA3A"
                     /translation="MEASPASGPRHLMDPHIFTSNFNNGIGRHKTYLCYEVERLDNGTS
                     VKMDQHRGFLHNQAKNLLCGFYGRHAELRFLDLVPSLQLDPAQIYRVTWFISWSPCFSW
                     GCAGEVRAFLQENTHVRLRIFAARIYDYDPLYKEALQMLRDAGAQVSIMTYDEFKHCWD
                     TFVDHQGCPFQPWDGLDEHSQALSGRLRAILQNQGN"
     CDS             687..707
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             720..743
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     primer_bind     complement(796..814)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      810..857
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 acaacaacaa taacaataac aacaacctcg ggatcgaggg aaggatttca gaattcgaaa
       61 acctgtattt tcaggggtcg acaatggaag ccagcccagc atccgggccc agacacttga
      121 tggatccaca catattcact tccaacttta acaatggcat tggaaggcat aagacctacc
      181 tgtgctacga agtggagcgc ctggacaatg gcacctcggt caagatggac cagcacaggg
      241 gctttctaca caaccaggct aagaatcttc tctgtggctt ttacggccgc catgcggagc
      301 tgcgcttctt ggacctggtt ccttctttgc agttggaccc ggcccagatc tacagggtca
      361 cttggttcat ctcctggagc ccctgcttct cctggggctg tgccggggaa gtgcgtgcgt
      421 tccttcagga gaacacacac gtgagactgc gtatcttcgc tgcccgcatc tatgattacg
      481 accccctata taaggaggca ctgcaaatgc tgcgggatgc tggggcccaa gtctccatca
      541 tgacctacga tgaatttaag cactgctggg acacctttgt ggaccaccag ggatgtccct
      601 tccagccctg ggatggacta gatgagcaca gccaagccct gagtgggagg ctgcgggcca
      661 ttctccagaa tcagggaaac ggtaccgaaa acctgtattt tcagggctcg acaatggatc
      721 accaccacca ccaccaccac cactaaccta ggctgctaaa caaagcccga aaggaagctg
      781 agttggctgc tgccaccgct gagcaataac tagcataacc ccttggggcc tctaaacggg
      841 tcttgagggg ttttttgctg aaaggaggaa ctatatccgg attggcgaat gggacgcg
//
