LOCUS       sequence-375917-        5060 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Test the C-to-U editing activity of KP6:C93A/C96A on AtrpoA editing 
            site, in human cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5060)
  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 5060)
  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-KP6-C93A-C96A_rpoA-C
FEATURES             Location/Qualifiers
     source          1..5060
                     /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    3011..3132
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3022..3041)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3076..3095
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3269..3373
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3374..3442,3443..4234)
                     /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(3592..3611)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4405..4993
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4894..4913
                     /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 acgttgtggc
     2041 ttggaactca atgataaaag gttatggaat gaaccatgaa ggcaaaatgg ccgtccaatg
     2101 tttcgaagat atgcaaaaac aaggtgtaaa gccttccgcc gtcactttta cctgtttgct
     2161 gaccgcctgt tctcacgcca gcctcgtcag cgaagggcag gagtatttta aaatgatgcg
     2221 agaagagtac gggatcgctc cgagggtgga acattacggc tgcatggtcg atctgcttgc
     2281 tcggagtggc catctgtatg aagctgaaaa gttcctggag atgctctgtc ctcccaacga
     2341 agggacatgg ggggccctcc tcagcgcttg taagacttat ggcgaggtgg agcttggtct
     2401 gaggtgcttt cagcagctcg tgcagcttaa cccagaaagt gccgcctggt acgtgcttat
     2461 ggctgacata tacgctggcg ccgggcggtg ggacgacgca taccggatcg aagagctgag
     2521 gaaacatgcc ggagccaaga agaaacctgc ctccgcattg attgaagtga acaaaaaagt
     2581 gcacgaattt gtggtcggca ataatcagag cgaggagatc agtgcgatgc ttaagtctct
     2641 caactctaga atgaaggagg agggccatgt acccaatctg gatctggttc tcaaacccgt
     2701 gtcagatgag gagaaagagg cagcgctgtg tgagcacgca gagaagttgg caattgcatt
     2761 cggcttgctg aatacgccac agggacagac cctgcgcgtt acaaagaacc tgagaatggc
     2821 caatgacgcc cacaatgcgt caaagatcat ctccaagatt gagcgcagag agattatcct
     2881 gcgcgatgat tgctgcattc accacttcaa ggacggactg tgctcctgcg gagacatgtt
     2941 ctaactcgag aaatagaagg aacatgtatt acacgtgcaa aatctgagag ctagcagcga
     3001 gcagacatga taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa
     3061 aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc
     3121 aataaacaag ttaacaacaa caattgcact ggttcatttt atgtttcagg ttcaggggga
     3181 ggtgtgggag gttttttaaa gcaagtaaaa cctctacaaa tgtggtaaaa tcgataagga
     3241 tccaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     3301 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     3361 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     3421 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     3481 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     3541 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     3601 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     3661 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     3721 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     3781 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     3841 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     3901 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     3961 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     4021 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     4081 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     4141 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     4201 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     4261 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     4321 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     4381 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     4441 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     4501 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     4561 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     4621 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     4681 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     4741 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     4801 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     4861 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     4921 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     4981 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     5041 ttttgctcac atggctcgac
//
