LOCUS       sequence-375926-        5066 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Test the U-to-C editing activity of KP4 on AtrpoA editing site, in 
            human cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5066)
  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 5066)
  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-KP4_rpoA-U
FEATURES             Location/Qualifiers
     source          1..5066
                     /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"
     polyA_signal    3017..3138
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3028..3047)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3082..3101
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3275..3379
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3380..3448,3449..4240)
                     /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(3598..3617)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4411..4999
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4900..4919
                     /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 acgttgtctc
     2041 ttggaatgca atgattgccg gttatgccca atgcgggaat attgaattgg ctcggcaatg
     2101 tttcgaagat atgcaacagg aaggcatcaa accaagcgag gtgactttca cctcaatact
     2161 tgctgcctgc tctcatgccg gtctggtcga cgaaggctgt aaatacttca agtccatgaa
     2221 agaggatcac gggataatac caaggacaca ccactacggc tgtatggttg accttctggg
     2281 ccgcgccggt taccttaatg aggccgaaaa tctcctgcag acgatccagc ctaacatcat
     2341 tggatggggg agccttttga ctgcttgtcg cacctatggg aatgtagagc tgggccgacg
     2401 ctgtttcgat cagatcgttc agttggaccc ccatagcgca tccggctatg tcctgatgtc
     2461 cggcatttat agcgacgctc agatgtggga ggatgccgca aaaatagagg agctcaggca
     2521 gcatgccggg gcttggaaga agcctgcaaa agcatggatc gaagtgaaca acaaagtgca
     2581 tgagtttatc gtgggcgata aaagccaccc ccagtccgag gaaatctatg ccatgctcaa
     2641 gaagctcagt aaccggatga aggaagaggg atacgtaccc aagatcgacc tggtgctgca
     2701 gcagatgtct gacgaagata aagaggatgc actctgcggg cattgcgaga aactggccat
     2761 cgcgtttgga ctgatttcta cccctccggg aacaaccatt cgggcgacaa agaacctgag
     2821 ggtgtgcatg gactgtcaca acgctactaa gatcatctca aagatcgaga agagagaaat
     2881 tattattaga gatgcttaca gactgcacca ctttaaggac ggatcatgca gttgcggaga
     2941 ctactactaa ctcgagaaat agaaggaaca tgtattacac gtgcaaaatt tgagagctag
     3001 cagcgagcag acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag
     3061 tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata
     3121 agctgcaata aacaagttaa caacaacaat tgcactggtt cattttatgt ttcaggttca
     3181 gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg gtaaaatcga
     3241 taaggatcca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     3301 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     3361 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     3421 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     3481 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     3541 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     3601 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     3661 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     3721 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     3781 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     3841 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     3901 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
     3961 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     4021 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     4081 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     4141 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     4201 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     4261 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     4321 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     4381 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     4441 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     4501 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     4561 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     4621 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     4681 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     4741 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     4801 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     4861 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     4921 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     4981 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     5041 ctggcctttt gctcacatgg ctcgac
//
