LOCUS       sequence-260783-        4739 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses APOBEC1 fused to the YTH domain.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4739)
  AUTHORS   Meyer KD
  TITLE     DART-seq: an antibody-free method for global m(6)A detection.
  JOURNAL   Nat Methods. 2019 Sep 23. pii: 10.1038/s41592-019-0570-0. doi: 
            10.1038/s41592-019-0570-0.
  PUBMED    31548708
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: "Nat 
            Methods. 2019 Sep 23. pii: 10.1038/s41592-019-0570-0. doi: 
            10.1038/s41592-019-0570-0."
COMMENT     Sequence Label: pCMV-APOBEC1-YTH
FEATURES             Location/Qualifiers
     source          1..4739
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        71..450
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        451..654
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     605..625
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     696..715
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        696..714
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             728..1414
                     /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             2051..2077
                     /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             2051..2077
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(2113..2130)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2119..2343
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(2414..2430)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2414..2430)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2427..2449)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2438..2454
                     /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(2462..2468,2469..2486,2487..2492))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2507..2528
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2645..2662)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2816..3404)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2896..2915)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3575..4366,4367..4435))
                     /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     4198..4217
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4436..4540)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4619..4638)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 tgcatgaaga atctgcttag ggttaggcgt tttgcgctgc ttcgcgatgt acgggccaga
       61 tatacgcgtt gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta
      121 gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc
      181 tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg
      241 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg
      301 gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa
      361 tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac
      421 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg
      481 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg
      541 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca
      601 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctggttta
      661 gtgaaccgtc agatccgcta gagatccgcg gccgctaata cgactcacta tagggagagc
      721 cgccaccatg agctcagaga ctggcccagt ggctgtggac cccacattga gacggcggat
      781 cgagccccat gagtttgagg tattcttcga tccgagagag ctccgcaagg agacctgcct
      841 gctttacgaa attaattggg ggggccggca ctccatttgg cgacatacat cacagaacac
      901 taacaagcac gtcgaagtca acttcatcga gaagttcacg acagaaagat atttctgtcc
      961 gaacacaagg tgcagcatta cctggtttct cagctggagc ccatgcggcg aatgtagtag
     1021 ggccatcact gaattcctgt caaggtatcc ccacgtcact ctgtttattt acatcgcaag
     1081 gctgtaccac cacgctgacc cccgcaatcg acaaggcctg cgggatttga tctcttcagg
     1141 tgtgactatc caaattatga ctgagcagga gtcaggatac tgctggagaa actttgtgaa
     1201 ttatagcccg agtaatgaag cccactggcc taggtatccc catctgtggg tacgactgta
     1261 cgttcttgaa ctgtactgca tcatactggg cctgcctcct tgtctcaaca ttctgagaag
     1321 gaagcagcca cagctgacat tctttaccat cgctcttcag tcttgtcatt accagcgact
     1381 gcccccacac attctctggg ccaccgggtt gaaaagcggc agcgagactc ccgggacctc
     1441 agagtccgcc acaccagaac cccacccagt gttggagaag cttcggtcca ttaataacta
     1501 taaccccaaa gattttgact ggaatctgaa acatggccgg gttttcatca ttaagagcta
     1561 ctctgaggac gatattcacc gttccattaa gtataatatt tggtgcagca cagagcatgg
     1621 taacaagaga ctggatgctg cttatcgttc catgaacggg aaaggccccg tttacttact
     1681 tttcagtgtc aacggcagtg gacacttctg tggcgtggca gaaatgaaat ctgctgtgga
     1741 ctacaacaca tgtgcaggtg tgtggtccca ggacaaatgg aagggtcgtt ttgatgtcag
     1801 gtggattttt gtgaaggacg ttcccaatag ccaactgcga cacattcgcc tagagaacaa
     1861 cgagaataaa ccagtgacca actctaggga cactcaggaa gtgcctctgg aaaaggctaa
     1921 gcaggtgttg aaaattatag ccagctacaa gcacaccact tccatttttg atgacttctc
     1981 acactatgag aaacgccaag aggaagaaga aagtgttaaa aaggaacgtc aaggtcgtgg
     2041 gaaactcgag tacccctacg acgtgcccga ctacgcctga gtttaaaccc gaaacttaaa
     2101 cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc
     2161 ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg
     2221 aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg
     2281 acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta
     2341 tggcttctga ggcggaaaga accagctggg gctcgatacc gtcgacctct agctagagct
     2401 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     2461 acaacatacg agccggaagc ataaagtgta aagcctaggg tgcctaatga gtgagctaac
     2521 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     2581 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     2641 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     2701 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     2761 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     2821 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     2881 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     2941 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     3001 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     3061 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     3121 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     3181 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     3241 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     3301 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     3361 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     3421 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     3481 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     3541 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     3601 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     3661 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     3721 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     3781 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     3841 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     3901 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     3961 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     4021 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     4081 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     4141 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     4201 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     4261 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     4321 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     4381 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     4441 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     4501 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     4561 cacctgacgt cgacggatcg ggagatcgat ctcccgatcc cctagggtcg actctcagta
     4621 caatctgctc tgatgccgca tagttaagcc agtatctgct ccctgcttgt gtgttggagg
     4681 tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg accgacaat
//
