LOCUS       sequence-375929-        5063 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Test the U-to-C editing activity of KP3: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 5063)
  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 5063)
  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: "Commun 
            Biol."; date: "2022-09-15"; volume: "5(1):968. doi"; pages: " 
            10.1038/s42003-022-03927-3"
COMMENT     Sequence Label: PM18033-PLS-KP3-C93A-C96A_rpoA-U
FEATURES             Location/Qualifiers
     source          1..5063
                     /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    3014..3135
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     3079..3098
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3272..3376
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3377..3445,3446..4237)
                     /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(3595..3614)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4408..4996
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4897..4916
                     /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 acgtcgttac
     2041 ttggaacgcc ctgatcagcg gttacggcac ctgtgatgag ggccacaaag ccctggcgtg
     2101 cttcgaggaa atgctcgtgg cggggatcca acctagcgac gctactttca cctgccttct
     2161 ggttgcctgt agtcatgagg gccttgtcca cgagggccgc aagcacttcg atagcatggc
     2221 cgaacaacat ggcataaagc ctcgcatcga acattatggt tgcatgatcg atctcctggg
     2281 aagagctggg caccttgatg aagctgagaa aatgctgctc agctgtgaag ggcagtccaa
     2341 tgtggtgggt ctgggcagcc tcctgaacgc ctgtaaatct cacggcgata tagagagagc
     2401 catgtggtgt tttgaatcta tcgtgcggct tgatcctcag tctgcctccg cctacgtact
     2461 gatggccaat gcctatgcgg acgccggccg ctggactgac gttgatagaa tcgaaaccca
     2521 gaggatggtg gctggcgcaa ataaaaagcc ggctgtagct accattgaag tgaacaaaca
     2581 ggtgcataga tttttggtcg gagaaaggag ggaggacatt gctctgaagg tggaatctac
     2641 gaatgcacga ctgaaacagg aggggggtca cgtcccccat acaaaacttg tgttgaagcc
     2701 cctctgtgag aaggagaagg aggacgcgct ctgtggacat gcagagaagc tggcactggc
     2761 attcggactg ctgaacacac ccagtgggac accactcgtg gttaccaaga acttgagaat
     2821 ggcctccgac gcccactcct caacagagat tatgtcaagg ttggagcagc gcgacattgt
     2881 cgtccgggac gggtaccggg tacaccgatt tgcagctgga tgctgctcat gcaagggacg
     2941 gccataactc gagaaataga aggaacatgt attacacgtg caaaatttga gagctagcag
     3001 cgagcagaca tgataagata cattgatgtg tttggacaaa ccacaactag aatgcagtga
     3061 aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc
     3121 tgcaataaac aagttaacaa caacaattgc actggttcat tttatgtttc aggttcaggg
     3181 ggaggtgtgg gaggtttttt aaagcaagta aaacctctac aaatgtggta aaatcgataa
     3241 ggatccaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     3301 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     3361 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     3421 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     3481 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     3541 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     3601 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     3661 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     3721 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     3781 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     3841 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     3901 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     3961 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     4021 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     4081 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     4141 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     4201 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     4261 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     4321 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     4381 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     4441 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     4501 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     4561 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     4621 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     4681 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     4741 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     4801 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     4861 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     4921 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     4981 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     5041 gccttttgct cacatggctc gac
//
