LOCUS       sequence-382432-        4739 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  APOBEC1 fused to YTHD422N.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4739)
  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 4739)
  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-APOBEC1-YTHD422N
FEATURES             Location/Qualifiers
     source          1..4739
                     /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             409..1095
                     /codon_start=1
                     /product="cytidine deaminase (C to U editing enzyme) from 
                     rat"
                     /label=APOBEC-1
                     /note="can use ssDNA as a substrate (Komor et al., 2016)"
                     /translation="MSSETGPVAVDPTLRRRIEPHEFEVFFDPRELRKETCLLYEINWG
                     GRHSIWRHTSQNTNKHVEVNFIEKFTTERYFCPNTRCSITWFLSWSPCGECSRAITEFL
                     SRYPHVTLFIYIARLYHHADPRNRQGLRDLISSGVTIQIMTEQESGYCWRNFVNYSPSN
                     EAHWPRYPHLWVRLYVLELYCIILGLPPCLNILRRKQPQLTFFTIALQSCHYQRLPPHI
                     LWATGLK"
     CDS             1732..1758
                     /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             1732..1758
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(1794..1811)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1800..2024
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(2095..2111)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2095..2111)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2108..2130)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2119..2135
                     /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(2143..2149,2150..2167,2168..2173))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2188..2209
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2326..2343)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2497..3085)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2577..2596)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3256..4047,4048..4116))
                     /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     3879..3898
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4117..4221)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4300..4319)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        join(4491..4739,1..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 gagctcagag
      421 actggcccag tggctgtgga ccccacattg agacggcgga tcgagcccca tgagtttgag
      481 gtattcttcg atccgagaga gctccgcaag gagacctgcc tgctttacga aattaattgg
      541 gggggccggc actccatttg gcgacataca tcacagaaca ctaacaagca cgtcgaagtc
      601 aacttcatcg agaagttcac gacagaaaga tatttctgtc cgaacacaag gtgcagcatt
      661 acctggtttc tcagctggag cccatgcggc gaatgtagta gggccatcac tgaattcctg
      721 tcaaggtatc cccacgtcac tctgtttatt tacatcgcaa ggctgtacca ccacgctgac
      781 ccccgcaatc gacaaggcct gcgggatttg atctcttcag gtgtgactat ccaaattatg
      841 actgagcagg agtcaggata ctgctggaga aactttgtga attatagccc gagtaatgaa
      901 gcccactggc ctaggtatcc ccatctgtgg gtacgactgt acgttcttga actgtactgc
      961 atcatactgg gcctgcctcc ttgtctcaac attctgagaa ggaagcagcc acagctgaca
     1021 ttctttacca tcgctcttca gtcttgtcat taccagcgac tgcccccaca cattctctgg
     1081 gccaccgggt tgaaaagcgg cagcgagact cccgggacct cagagtccgc cacaccagaa
     1141 ccccacccag tgttggagaa gcttcggtcc attaataact ataaccccaa agattttgac
     1201 tggaatctga aacatggccg ggttttcatc attaagagct actctgagga caatattcac
     1261 cgttccatta agtataatat ttggtgcagc acagagcatg gtaacaagag actggatgct
     1321 gcttatcgtt ccatgaacgg gaaaggcccc gtttacttac ttttcagtgt caacggcagt
     1381 ggacacttct gtggcgtggc agaaatgaaa tctgctgtgg actacaacac atgtgcaggt
     1441 gtgtggtccc aggacaaatg gaagggtcgt tttgatgtca ggtggatttt tgtgaaggac
     1501 gttcccaata gccaactgcg acacattcgc ctagagaaca acgagaataa accagtgacc
     1561 aactctaggg acactcagga agtgcctctg gaaaaggcta agcaggtgtt gaaaattata
     1621 gccagctaca agcacaccac ttccattttt gatgacttct cacactatga gaaacgccaa
     1681 gaggaagaag aaagtgttaa aaaggaacgt caaggtcgtg ggaaactcga gtacccctac
     1741 gacgtgcccg actacgcctg agtttaaacc cgaaacttaa acccgctgat cagcctcgac
     1801 tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct
     1861 ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct
     1921 gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg
     1981 ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag
     2041 aaccagctgg ggctcgatac cgtcgacctc tagctagagc ttggcgtaat catggtcata
     2101 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     2161 cataaagtgt aaagcctagg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     2221 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     2281 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     2341 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     2401 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     2461 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     2521 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     2581 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     2641 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     2701 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     2761 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     2821 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     2881 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     2941 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     3001 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     3061 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     3121 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     3181 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     3241 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     3301 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     3361 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     3421 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     3481 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     3541 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     3601 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     3661 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     3721 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     3781 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     3841 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     3901 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     3961 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     4021 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     4081 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     4141 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     4201 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc
     4261 gggagatcga tctcccgatc ccctagggtc gactctcagt acaatctgct ctgatgccgc
     4321 atagttaagc cagtatctgc tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag
     4381 caaaatttaa gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga atctgcttag
     4441 ggttaggcgt tttgcgctgc ttcgcgatgt acgggccaga tatacgcgtt gacattgatt
     4501 attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga
     4561 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
     4621 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
     4681 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatc
//
