LOCUS       sequence-382434-        5246 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ADAR1 catalytic domain E488Q fused to YTHD422N.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5246)
  AUTHORS   Zhu H, Yin X, Holley CL, Meyer KD
  TITLE     Improved Methods for Deamination-Based m(6)A Detection.
  JOURNAL   Front Cell Dev Biol. 2022 Apr 27;10:888279. doi: 
            10.3389/fcell.2022.888279. eCollection 2022.
  PUBMED    35573664
REFERENCE   2  (bases 1 to 5246)
  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: "Front Cell 
            Dev Biol."; date: "2022-04-27"; volume: "10:888279. doi"; pages: " 
            10.3389/fcell.2022.888279. eCollection 2022"
COMMENT     Sequence Label: pCMV-ADARcd-YTHD422N
FEATURES             Location/Qualifiers
     source          1..5246
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        132..335
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     286..306
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     377..396
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        377..395
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2239..2265
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YPYDVPDYA"
     CDS             2239..2265
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(2301..2318)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2307..2531
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(2602..2618)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2602..2618)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2615..2637)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2626..2642
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(2650..2656,2657..2674,2675..2680))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2695..2716
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2833..2850)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3004..3592)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3084..3103)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3763..4554,4555..4623))
                     /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     4386..4405
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4624..4728)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4807..4826)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        join(4998..5246,1..131)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
       61 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
      121 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
      181 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
      241 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
      301 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct
      361 agagatccgc ggccgctaat acgactcact atagggagag ccgccaccat gccatctcgc
      421 cagcctattc ccagtgaggg tcttcagctg catttaccgc aggttttagc tgacgctgtc
      481 tcacgcctgg tcctgggtaa gtttggtgac ctgaccgaca acttctcctc ccctcacgct
      541 cgcagaaaag tgctggctgg agtcgtcatg acaacaggca cagatgttaa agatgccaag
      601 gtgataagtg tttctacagg aacaaaatgt attaatggtg aatacatgag tgatcgtggc
      661 cttgcattaa atgactgcca tgcagaaata atatctcgga gatccttgct cagatttctt
      721 tatacacaac ttgagcttta cttaaataac aaagatgatc aaaaaagatc catctttcag
      781 aaatcagagc gaggggggtt taggctgaag gagaatgtcc agtttcatct gtacatcagc
      841 acctctccct gtggagatgc cagaatcttc tcaccacatg agccaatcct ggaagaacca
      901 gcagatagac acccaaatcg taaagcaaga ggacagctac ggaccaaaat agagtctggt
      961 caggggacga ttccagtgcg ctccaatgcg agcatccaaa cgtgggacgg ggtgctgcaa
     1021 ggggagcggc tgctcaccat gtcctgcagt gacaagattg cacgctggaa cgtggtgggc
     1081 atccagggat ccctgctcag cattttcgtg gagcccattt acttctcgag catcatcctg
     1141 ggcagccttt accacgggga ccacctttcc agggccatgt accagcggat ctccaacata
     1201 gaggacctgc cacctctcta caccctcaac aagcctttgc tcagtggcat cagcaatgca
     1261 gaagcacggc agccagggaa ggcccccaac ttcagtgtca actggacggt aggcgactcc
     1321 gctattgagg tcatcaacgc cacgactggg aaggatgagc tgggccgcgc gtcccgcctg
     1381 tgtaagcacg cgttgtactg tcgctggatg cgtgtgcacg gcaaggttcc ctcccactta
     1441 ctacgctcca agattaccaa acccaacgtg taccatgagt ccaagctggc ggcaaaggag
     1501 taccaggccg ccaaggcgcg tctgttcaca gccttcatca aggcggggct gggggcctgg
     1561 gtggagaagc ccaccgagca ggaccagttc tcactcacgc ccagcggcag cgagactccc
     1621 gggacctcag agtccgccac accagaaccc cacccagtgt tggagaagct tcggtccatt
     1681 aataactata accccaaaga ttttgactgg aatctgaaac atggccgggt tttcatcatt
     1741 aagagctact ctgaggacaa tattcaccgt tccattaagt ataatatttg gtgcagcaca
     1801 gagcatggta acaagagact ggatgctgct tatcgttcca tgaacgggaa aggccccgtt
     1861 tacttacttt tcagtgtcaa cggcagtgga cacttctgtg gcgtggcaga aatgaaatct
     1921 gctgtggact acaacacatg tgcaggtgtg tggtcccagg acaaatggaa gggtcgtttt
     1981 gatgtcaggt ggatttttgt gaaggacgtt cccaatagcc aactgcgaca cattcgccta
     2041 gagaacaacg agaataaacc agtgaccaac tctagggaca ctcaggaagt gcctctggaa
     2101 aaggctaagc aggtgttgaa aattatagcc agctacaagc acaccacttc catttttgat
     2161 gacttctcac actatgagaa acgccaagag gaagaagaaa gtgttaaaaa ggaacgtcaa
     2221 ggtcgtggga aactcgagta cccctacgac gtgcccgact acgcctgagt ttaaacccga
     2281 aacttaaacc cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg
     2341 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata
     2401 aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt
     2461 ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt
     2521 gggctctatg gcttctgagg cggaaagaac cagctggggc tcgataccgt cgacctctag
     2581 ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     2641 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctagggtg cctaatgagt
     2701 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     2761 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     2821 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     2881 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     2941 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     3001 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     3061 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     3121 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     3181 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     3241 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     3301 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     3361 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     3421 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     3481 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     3541 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     3601 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     3661 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     3721 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     3781 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     3841 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     3901 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     3961 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     4021 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     4081 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     4141 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     4201 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     4261 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     4321 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     4381 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     4441 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     4501 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     4561 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     4621 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     4681 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     4741 aaaagtgcca cctgacgtcg acggatcggg agatcgatct cccgatcccc tagggtcgac
     4801 tctcagtaca atctgctctg atgccgcata gttaagccag tatctgctcc ctgcttgtgt
     4861 gttggaggtc gctgagtagt gcgcgagcaa aatttaagct acaacaaggc aaggcttgac
     4921 cgacaattgc atgaagaatc tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg
     4981 ggccagatat acgcgttgac attgattatt gactagttat taatagtaat caattacggg
     5041 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
     5101 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
     5161 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
     5221 ccacttggca gtacatcaag tgtatc
//
