LOCUS       sequence-375913-        5057 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Test the U-to-C editing activity of KP2:A66S on AtrpoA editing site,
            in human cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5057)
  AUTHORS   Ichinose M, Kawabata M, Akaiwa Y, Shimajiri Y, Nakamura I, Tamai T, 
            Nakamura T, Yagi Y, Gutmann B
  TITLE     U-to-C RNA editing by synthetic PPR-DYW proteins in bacteria and 
            human culture cells.
  JOURNAL   Commun Biol. 2022 Sep 15;5(1):968. doi: 10.1038/s42003-022-03927-3.
  PUBMED    36109586
REFERENCE   2  (bases 1 to 5057)
  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: "Commun 
            Biol."; date: "2022-09-15"; volume: "5(1):968. doi"; pages: " 
            10.1038/s42003-022-03927-3"
COMMENT     Sequence Label: PM18033-PLS-KP2-A66S_rpoA-U
FEATURES             Location/Qualifiers
     source          1..5057
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    152..163
                     /label=ZF binding site
                     /bound_moiety="CCR5TC zinc-finger domain"
                     /note="target sequence for the CCR5TC zinc-finger domain 
                     from the CCR5-224 (+) zinc-finger nuclease (Perez et al., 
                     2008; Gross et al., 2013)"
     enhancer        171..550
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        551..754
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     705..725
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     751..775
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          890..1022
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1005..1024
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             1061..1078
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1088..1108
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             2911..2922
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     polyA_signal    3008..3129
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3019..3038)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3073..3092
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3266..3370
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3371..3439,3440..4231)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3589..3608)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4402..4990
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4891..4910
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agatcttcaa tattggccat tagccatatt attcattggt tatatagcat aaatcaatat
       61 tggctattgg ccattgcata cgttgtatct atatcataat atgtacattt atattggctc
      121 atgtccaata tgaccgccat gttgtcttag agtcatcctc atcgtcggac ggcattgatt
      181 attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga
      241 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
      301 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
      361 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
      421 tatgccaagt ccgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
      481 ccagtacatg accttacggg actttcctac ttggcagtac atctacgtat tagtcatcgc
      541 tattaccatg gtgatgcggt tttggcagta caccaatggg cgtggatagc ggtttgactc
      601 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa
      661 tcaacgggac tttccaaaat gtcgtaataa ccccgccccg ttgacgcaaa tgggcggtag
      721 gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc agatcactag
      781 aagctttatt gcggtagttt atcacagtta aattgctaac gcagtcagtg cttctgacac
      841 aacagtctcg aacttaagct gcagaagttg gtcgtgaggc actgggcagg taagtatcaa
      901 ggttacaaga caggtttaag gagaccaata gaaactgggc ttgtcgagac agagatgact
      961 cttgcgtttc tgataggcac ctattggtct tactgacatc cactttgcct ttctctccac
     1021 aggtgtccac tcccagttca attacacacg caccatggcg caccatcatc atcatcattc
     1081 ttctggtgag aatctttatt ttcagggcgc catgggtaat tctgttgtca catggaattc
     1141 catgattagg ggctacgcta gaagcggcca gcccgaggaa gctctgagcc tcttctccca
     1201 gatgagaagg agcggcatca agcccgatag ctataccttc cccttcgtgc tgaaggcttg
     1261 cgcttctctg agcagcctcg aggaaggaaa acaaatccac gcccacgtca ttaaatccgg
     1321 cttcgagtcc gatgtctacg tgcagagctc cctcattgat atgtacgcca agtgcggatc
     1381 tctggaggac gccagaaaag tgtttgacga gatgcccgag aggaatgtcg tgagctggaa
     1441 cgctatgatc agcggctatg cccagaatgg acaatccgaa gaagctctgg agctgtttag
     1501 agagatgcag aaagagggca tcaagccttc cgagtttacc ttttgctccg tgctgaaggc
     1561 ttgcgcctct ctgggatctc tggagatggg caagcaaatt cacggctatg tgatcaagtc
     1621 cggctttgag tccatcgtgt tcgtgggaaa cgccctcatt gacatgtatg ccaagtgcgg
     1681 ctctctggag gatgctagaa aggtctttga tgagatgccc gaaaggaccg ttgtgagctg
     1741 gaccgccatg atctccggat acgcccagaa cggacagagc gaggaggccc tcgagctctt
     1801 cagagagatg cagagggagg gcgtgaaacc cgacgaagtg acactgcctt ccgtgctctc
     1861 cgcttgcgcc aatctgggcg ctctggagca aggcaaacag atccatgcct acgtcatcaa
     1921 gaacggcttt gaatccgatg tgtttgtggg ctccgctctg atcgacatgt acgctaaatg
     1981 cggctccatc gaggaggcta ggaaagtgtt cgatgaaatg cccgagaaag acgtggtgac
     2041 ttggaacgct atgatcaacg gttatggcca atctaacgaa gggcataaag ccatcgaatg
     2101 cttccaggag atgttgcagg acggggtgca gcctagcgag acaacattca catgtctcct
     2161 cgtcgcctgt tctcacgaag gcctcgtgaa cgaaggacgc caatacttta aggccatggt
     2221 ggaggaacac ggaatcatgc ctcgcgtgga acattacggg tgtatggttg acctgctcgg
     2281 tagggccggc tatctggatg aggccgaaaa gatgctgctg tcctttgaga gcaatgatgt
     2341 gggattcggt tccctcctta acgcttgccg ctcccacggc gacatagaac ggggcacacg
     2401 ctgttttgaa agcctggtcc gcctcgaccc tcagtctgct agcgcctacg tacttatggc
     2461 caatatatac gcagatgccg gtcggtggac tgatgtggac aggatcgaga gactgcggaa
     2521 gtcagcgggc gcatccaaaa aacccgcggt ggcaggcatc gaagtcgaca ggcacgtgca
     2581 cacctttgtc gtcggggaag aaaggggcga cattagctgt aaagtaaggt ctatgaatac
     2641 gcggctgaaa gaggagggag gccatgtgcc gcataccgag ctggtgctta aaccactgtc
     2701 cgagaaggag aaggaggacg agctgtgcgg gcattcagag aagttggcac tggcttatgg
     2761 acttctgaat actcccgatg ggaccccctt gttggtcacc aagaatctgc ggatgtgcag
     2821 agactgccac agttcaaccg agattatgtc aagaattgag aagcgagaga ttgttgttag
     2881 agatgcacac cgagttcaca gattcgttga tgggagttgc agttgcggag gacgaccata
     2941 actcgagaaa tagaaggaac atgtattaca cgtgcaaaat ttgagagcta gcagcgagca
     3001 gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa
     3061 tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat
     3121 aaacaagtta acaacaacaa ttgcactggt tcattttatg tttcaggttc agggggaggt
     3181 gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt ggtaaaatcg ataaggatcc
     3241 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     3301 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     3361 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     3421 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     3481 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     3541 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     3601 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     3661 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     3721 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     3781 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     3841 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     3901 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     3961 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     4021 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     4081 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     4141 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     4201 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     4261 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     4321 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     4381 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     4441 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     4501 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     4561 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     4621 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     4681 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     4741 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     4801 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     4861 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     4921 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     4981 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     5041 tgctcacatg gctcgac
//
