LOCUS       sequence-382431-        5164 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ADAR1 catalytic domain E488Q fused to YTHmut.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5164)
  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 5164)
  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-YTHmut
FEATURES             Location/Qualifiers
     source          1..5164
                     /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             2164..2190
                     /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             2164..2190
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(2219..2236)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2225..2449
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(2520..2536)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2520..2536)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2533..2555)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2544..2560
                     /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(2568..2574,2575..2592,2593..2598))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2613..2634
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2751..2768)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2922..3510)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3002..3021)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3681..4472,4473..4541))
                     /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     4304..4323
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4542..4646)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4725..4744)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        join(4916..5164,1..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 accagaaggc cgggttttca tcattaagag ctactctgag
     1681 gacgatattc accgttccat taagtataat atttggtgca gcacagagca tggtaacaag
     1741 agactggatg ctgcttatcg ttccatgaac gggaaaggcc ccgtttactt acttttcagt
     1801 gtcaacggca gtggacactt ctgtggcgtg gcagaaatga aatctgctgt ggactacaac
     1861 acatgtgcag gtgtgtggtc ccaggacaaa tggaagggtc gttttgatgt caggtggatt
     1921 tttgtgaagg acgttcccaa tagccaactg cgacacattc gcctagagaa caacgagaat
     1981 aaaccagtga ccaactctag ggacactcag gaagtgcctc tggaaaaggc taagcaggtg
     2041 ttgaaaatta tagccagcta caagcacacc acttccattt ttgatgactt ctcacactat
     2101 gagaaacgcc aagaggaaga agaaagtgtt aaaaaggaac gtcaaggtcg tgggaaactc
     2161 gagtacccct acgacgtgcc cgactacgcc tgagtttaaa cttaaacccg ctgatcagcc
     2221 tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg
     2281 accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat
     2341 tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag
     2401 gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg
     2461 gaaagaacca gctggggctc gataccgtcg acctctagct agagcttggc gtaatcatgg
     2521 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     2581 ggaagcataa agtgtaaagc ctagggtgcc taatgagtga gctaactcac attaattgcg
     2641 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     2701 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     2761 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     2821 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     2881 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     2941 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     3001 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     3061 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     3121 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     3181 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     3241 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     3301 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     3361 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     3421 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     3481 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     3541 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     3601 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     3661 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     3721 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     3781 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     3841 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     3901 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     3961 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     4021 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     4081 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     4141 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     4201 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     4261 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     4321 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     4381 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     4441 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     4501 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     4561 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     4621 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac
     4681 ggatcgggag atcgatctcc cgatccccta gggtcgactc tcagtacaat ctgctctgat
     4741 gccgcatagt taagccagta tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc
     4801 gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg acaattgcat gaagaatctg
     4861 cttagggtta ggcgttttgc gctgcttcgc gatgtacggg ccagatatac gcgttgacat
     4921 tgattattga ctagttatta atagtaatca attacggggt cattagttca tagcccatat
     4981 atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac
     5041 ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc
     5101 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg
     5161 tatc
//
