LOCUS       sequence-247870-        3285 bp    DNA     circular SYN 15-APR-2025
DEFINITION  clone A3A-W98Y 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 Apr 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: pBT271.361
FEATURES             Location/Qualifiers
     source          1..3285
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             461..472
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     protein_bind    490..508
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    515..533
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     592..609
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             624..1217
                     /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
                     KMDQHRGFLHNQAKNLLCGFYGRHAELRFLDLVPSLQLDPAQIYRVTWFISYSPCFSWG
                     CAGEVRAFLQENTHVRLRIFAARIYDYDPLYKEALQMLRDAGAQVSIMTYDEFKHCWDT
                     FVDHQGCPFQPWDGLDEHSQALSGRLRAILQNQGN"
     terminator      complement(1401..1435)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(1506..2165)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2080..2099
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      2347..2935
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2836..2855
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(3014..3045)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     CDS             complement(join(3062..3285,1..454))
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
ORIGIN
        1 ggtacatacg ttcggtggaa gcttcccaac ccatggtttt tttctgcata accggaccgt
       61 cggacgggaa gttggtacca cgcagtttaa ctttgtagat gaactcaccg tcttgcaggg
      121 aggagtcctg ggtaacggta acaacaccac cgtcttcgaa gttcataaca cgttcccatt
      181 tgaaaccttc cgggaaggac agtttcaggt agtccgggat gtcagccggg tgtttaacgt
      241 aagctttgga accgtactgg aactgcgggg acaggatgtc ccaagcgaac ggcagcggac
      301 cacctttggt aactttcagt ttagcggtct gggtaccttc gtacggacga ccttcacctt
      361 caccttcgat ttcgaactcg tgaccgttaa cggaaccttc catacgaact ttgaaacgca
      421 tgaactcttt gataacgtct tcgctactcg ccatagatcc tttctcctct ttcagatccg
      481 tgctcagtat ctctatcact gatagggatg tcaatctcta tcactgatag ggaagggtga
      541 ctcgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
      601 gtgagcgaga atgctcttct agtgaagcca gcccagcatc cgggcccaga cacttgatgg
      661 atccacacat attcacttcc aactttaaca atggcattgg aaggcataag acctacctgt
      721 gctacgaagt ggagcgcctg gacaatggca cctcggtcaa gatggaccag cacaggggct
      781 ttctacacaa ccaggctaag aatcttctct gtggctttta cggccgccat gcggagctgc
      841 gcttcttgga cctggttcct tctttgcagt tggacccggc ccagatctac agggtcactt
      901 ggttcatctc ctatagcccc tgcttctcct ggggctgtgc cggggaagtg cgtgcgttcc
      961 ttcaggagaa cacacacgtg agactgcgca tcttcgctgc ccgcatctat gattacgacc
     1021 ccctatataa ggaggcgctg caaatgctgc gggatgctgg ggcccaagtc tccatcatga
     1081 cctacgatga gttcaagcac tgctgggaca cctttgtgga ccaccaggga tgtcccttcc
     1141 agccctggga tggactagat gagcacagcc aagccctgag tgggaggctg cgggccattc
     1201 tccagaatca gggaaactct ggctgaagag cctggttaaa aaatgagctg atttaacaaa
     1261 aatttaacgc gaattttaac aaaatattaa cgcttacaat ttaggtggca cttttcgggg
     1321 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     1381 catgaattaa ttcttaacca ataaaaaacg cccggcggca accgagcgtt ctgaacaaat
     1441 ccagatggag ttctgaggtc attactggat ctatcaacag gagtccaagc gagctcgata
     1501 tcaaattacg ccccgccctg ccactcatcg cagtactgtt gtaattcatt aagcattctg
     1561 ccgacatgga agccatcaca aacggcatga tgaacctgaa tcgccagcgg catcagcacc
     1621 ttgtcgcctt gcgtataata tttgcccatg gtgaaaacgg gggcgaagaa gttgtccata
     1681 ttggccacgt ttaaatcaaa actggtgaaa ctcacccagg gattggctga aacgaaaaac
     1741 atattctcaa taaacccttt agggaaatag gccaggtttt caccgtaaca cgccacatct
     1801 tgcgaatata tgtgtagaaa ctgccggaaa tcgtcgtggt attcactcca gagcgatgaa
     1861 aacgtttcag tttgctcatg gaaaacggtg taacaagggt gaacactatc ccatatcacc
     1921 agctcaccgt ctttcattgc catacgaaat tccggatgag cattcatcag gcgggcaaga
     1981 atgtgaataa aggccggata aaacttgtgc ttatttttct ttacggtctt taaaaaggcc
     2041 gtaatatcca gctgaacggt ctggttatag gtacattgag caactgactg aaatgcctca
     2101 aaatgttctt tacgatgcca ttgggatata tcaacggtgg tatatccagt gatttttttc
     2161 tccattttag cttccttagc tcctgaaaat ctcgataact caaaaaatac gcccggtagt
     2221 gatcttattt cattatggtg aaagttggaa cctcttacgt gcccgatcaa tcatgaccaa
     2281 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     2341 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     2401 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     2461 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     2521 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     2581 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     2641 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     2701 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     2761 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     2821 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     2881 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     2941 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     3001 tactgcgtta tgtagtcaaa agcctccggt cggaggcttt tgacttggct gaggagtgcc
     3061 attaagcacc ggtggagtga cgaccttcag cacgttcgta ctgttcaacg atggtgtagt
     3121 cttcgttgtg ggaggtgatg tccagtttga tgtcggtttt gtaagcaccc ggcagctgaa
     3181 ccggtttttt agccatgtag gtggttttaa cttcagcgtc gtagtgacca ccgtctttca
     3241 gtttcagacg cattttgatt tcacctttca gagcaccgtc ttccg
//
