LOCUS       sequence-248970-        3285 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  clone A3A CBEs by SapI Golden Gate.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3285)
  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 3285)
  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: pBT271.359
FEATURES             Location/Qualifiers
     source          1..3285
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(642..676)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(747..1406)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1321..1340
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      1588..2176
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2077..2096
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2255..2286)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     CDS             complement(2303..2980)
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     RBS             2987..2998
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     protein_bind    3016..3034
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3041..3059
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     3118..3135
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             join(3150..3285,1..458)
                     /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="EASPASGPRHLMDPHIFTSNFNNGIGRHKTYLCYEVERLDNGTSV
                     KMDQHRGFLHNQAKNLLCGFYGRHAELRFLDLVPSLQLDPAQIYRVTWFISWSPCFSWG
                     CAGEVRAFLQENTHVRLRIFAARIYDYDPLYKEALQMLRDAGAQVSIMTYDEFKHCWDT
                     FVDHQGCPFQPWDGLDEHSQALSGRLRAILQNQGN"
ORIGIN
        1 agatggacca gcacaggggc tttctacaca accaggctaa gaatcttctc tgtggctttt
       61 acggccgcca tgcggagctg cgcttcttgg acctggttcc ttctttgcag ttggacccgg
      121 cccagatcta cagggtcact tggttcatct cctggagccc ctgcttctcc tggggctgtg
      181 ccggggaagt gcgtgcgttc cttcaggaga acacacacgt gagactgcgc atcttcgctg
      241 cccgcatcta tgattacgac cccctatata aggaggcgct gcaaatgctg cgggatgctg
      301 gggcccaagt ctccatcatg acctacgatg agttcaagca ctgctgggac acctttgtgg
      361 accaccaggg atgtcccttc cagccctggg atggactaga tgagcacagc caagccctga
      421 gtgggaggct gcgggccatt ctccagaatc agggaaactc tggctgaaga gcctggttaa
      481 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa
      541 tttaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
      601 acattcaaat atgtatccgc tcatgaatta attcttaacc aataaaaaac gcccggcggc
      661 aaccgagcgt tctgaacaaa tccagatgga gttctgaggt cattactgga tctatcaaca
      721 ggagtccaag cgagctcgat atcaaattac gccccgccct gccactcatc gcagtactgt
      781 tgtaattcat taagcattct gccgacatgg aagccatcac aaacggcatg atgaacctga
      841 atcgccagcg gcatcagcac cttgtcgcct tgcgtataat atttgcccat ggtgaaaacg
      901 ggggcgaaga agttgtccat attggccacg tttaaatcaa aactggtgaa actcacccag
      961 ggattggctg aaacgaaaaa catattctca ataaaccctt tagggaaata ggccaggttt
     1021 tcaccgtaac acgccacatc ttgcgaatat atgtgtagaa actgccggaa atcgtcgtgg
     1081 tattcactcc agagcgatga aaacgtttca gtttgctcat ggaaaacggt gtaacaaggg
     1141 tgaacactat cccatatcac cagctcaccg tctttcattg ccatacgaaa ttccggatga
     1201 gcattcatca ggcgggcaag aatgtgaata aaggccggat aaaacttgtg cttatttttc
     1261 tttacggtct ttaaaaaggc cgtaatatcc agctgaacgg tctggttata ggtacattga
     1321 gcaactgact gaaatgcctc aaaatgttct ttacgatgcc attgggatat atcaacggtg
     1381 gtatatccag tgattttttt ctccatttta gcttccttag ctcctgaaaa tctcgataac
     1441 tcaaaaaata cgcccggtag tgatcttatt tcattatggt gaaagttgga acctcttacg
     1501 tgcccgatca atcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     1561 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     1621 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     1681 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     1741 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     1801 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     1861 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     1921 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     1981 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     2041 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     2101 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     2161 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     2221 gccttttgct cacatgttct ttactgcgtt atgtagtcaa aagcctccgg tcggaggctt
     2281 ttgacttggc tgaggagtgc cattaagcac cggtggagtg acgaccttca gcacgttcgt
     2341 actgttcaac gatggtgtag tcttcgttgt gggaggtgat gtccagtttg atgtcggttt
     2401 tgtaagcacc cggcagctga accggttttt tagccatgta ggtggtttta acttcagcgt
     2461 cgtagtgacc accgtctttc agtttcagac gcattttgat ttcacctttc agagcaccgt
     2521 cttccgggta catacgttcg gtggaagctt cccaacccat ggtttttttc tgcataaccg
     2581 gaccgtcgga cgggaagttg gtaccacgca gtttaacttt gtagatgaac tcaccgtctt
     2641 gcagggagga gtcctgggta acggtaacaa caccaccgtc ttcgaagttc ataacacgtt
     2701 cccatttgaa accttccggg aaggacagtt tcaggtagtc cgggatgtca gccgggtgtt
     2761 taacgtaagc tttggaaccg tactggaact gcggggacag gatgtcccaa gcgaacggca
     2821 gcggaccacc tttggtaact ttcagtttag cggtctgggt accttcgtac ggacgacctt
     2881 caccttcacc ttcgatttcg aactcgtgac cgttaacgga accttccata cgaactttga
     2941 aacgcatgaa ctctttgata acgtcttcgc tactcgccat agatcctttc tcctctttca
     3001 gatccgtgct cagtatctct atcactgata gggatgtcaa tctctatcac tgatagggaa
     3061 gggtgactcg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
     3121 gagtcagtga gcgagaatgc tcttctagtg aagccagccc agcatccggg cccagacact
     3181 tgatggatcc acacatattc acttccaact ttaacaatgg cattggaagg cataagacct
     3241 acctgtgcta cgaagtggag cgcctggaca atggcacctc ggtca
//
