LOCUS       sequence-433759-        7664 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of FLAG-HA-tagged human YTHDC1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7664)
  AUTHORS   Patil DP, Chen CK, Pickering BF, Chow A, Jackson C, Guttman M, 
            Jaffrey SR
  TITLE     m6A RNA methylation promotes XIST-mediated transcriptional 
            repression.
  JOURNAL   Nature. 2016 Sep 7;537(7620):369-373. doi: 10.1038/nature19342.
  PUBMED    27602518
REFERENCE   2  (bases 1 to 7664)
  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"; doi: "10"; journalName: 
            "Nature"; date: "2016-09-7- 7"; volume: "537"; issue: "7620"; pages:
            "369-373"
COMMENT     Sequence Label: pcDNA3-FLAG-HA-hYTHDC1
FEATURES             Location/Qualifiers
     source          1..7664
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             901..924
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             937..963
                     /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             937..963
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        complement(3217..3235)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3218..3235)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3255..3272)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3261..3485
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3531..3959
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3618..3637)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3828..3849
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3968..3988)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3973..4302
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4153..4288
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4215..4234
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4369..5163
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4423..4442)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5033..5052
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5337..5458
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5374..5393)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5428..5447
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5507..5523)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5507..5523)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5520..5542)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5531..5547
                     /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(5555..5561,5562..5579,5580..5585))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5600..5621
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5738..5755)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5909..6497)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5989..6008)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6668..7459,7460..7528))
                     /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     7291..7310
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7529..7633)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttaccatg
      901 gactacaagg acgacgatga caagctcgat ggaggatacc cctacgacgt gcccgactac
      961 gccagcggat ccactagtaa cggccgccag tgtgctggaa ttctgatggc ggctgacagt
     1021 cgggaggaga aagatggaga acttaatgtt ctggatgata ttttaactga agtaccagaa
     1081 caagatgatg aactgtataa tccagagagt gaacaagata aaaatgagaa aaagggatca
     1141 aaaagaaaaa gtgatcgaat ggaatctact gataccaaac gacaaaagcc ttctgtccat
     1201 tcaagacaac tggtttctaa gccactgagc tcatctgtta gcaataacaa aagaatagtt
     1261 agtacaaaag gaaagtcagc cacagagtat aaaaatgagg aatatcaaag atctgaaaga
     1321 aacaagcgtc tagatgctga tcggaaaatt cgtctatcaa gtagtgcctc cagagaacct
     1381 tataagaatc aacctgaaaa aacctgtgtc cggaaaaggg atcctgaaag gagggccaaa
     1441 tctcctacgc cagatggttc tgagagaatt gggcttgaag tggatagacg tgcaagcaga
     1501 tccagccagt cttctaagga agaagtgaac tctgaagaat atggctctga ccatgagact
     1561 ggcagcagtg gttcttctga tgagcaaggg aacaacactg agaatgagga ggaaggagtg
     1621 gaagaagatg tggaggaaga tgaagaagta gaagaagatg cagaagaaga tgaagaggtg
     1681 gatgaagatg gagaggagga ggaggaagag gaggaggagg aagaggagga ggaggaggag
     1741 gaagaagaag aatatgaaca ggatgagaga gaccagaaag aggagggaaa tgattatgac
     1801 actcgaagtg aggccagtga ctctggttct gaatctgttt ccttcacaga tgggtctgtc
     1861 agatctggtt caggcacaga tggatcagat gagaaaaaga aggaaaggaa gagagctaga
     1921 ggcatatctc caattgtttt tgatagaagt ggaagctctg catcagagtc atatgcaggt
     1981 tcagaaaaga agcatgagaa attatcatct tccgttcgtg ctgtccgaaa agatcaaacc
     2041 agtaaactca aatatgtgct tcaagatgca agatttttcc tcataaagag taacaaccat
     2101 gagaatgtgt ctcttgccaa agcgaagggt gtatggtcca cgctccctgt aaatgagaag
     2161 aaattaaatc ttgcatttag atctgcaagg agtgttatct taatattttc tgtcagagag
     2221 agtggaaaat ttcaagggtt tgcaagactt tcttcagaat cacatcacgg aggatctcct
     2281 atacactggg tgcttccagc aggaatgagt gctaaaatgc tgggaggtgt ctttaaaatt
     2341 gactggattt gcaggcgtga attacccttc actaagtcgg ctcatctcac caatccttgg
     2401 aatgaacata aaccagtaaa gatcggacgt gatggacagg aaattgaact tgaatgtgga
     2461 acccagcttt gtcttctgtt tccccccgat gaaagtattg acttgtatca ggtcattcat
     2521 aaaatgcgtc acaagagaag aatgcattct cagccccgat cacgaggacg tccatcccgt
     2581 cgagaaccag tccgggatgt gggaaggcgt cgaccagaag attatgatat tcataacagc
     2641 agaaagaaac caaggattga ctatccccct gagtttcacc agagaccagg gtatttaaag
     2701 gatccacgat accaggaagt ggacagacga ttttcaggag ttcgccgaga tgtgttttta
     2761 aatgggtcct acaatgatta tgtgagggaa tttcataaca tgggaccacc accaccttgg
     2821 caaggaatgc ccccttaccc aggaatggaa caacctccac accatcctta ctatcagcac
     2881 catgctccac ctcctcaagc tcatccccct tactcaggac atcatccagt accacatgaa
     2941 gcaagataca gagataaacg agtacatgat tatgatatga gggtggatga tttccttcgt
     3001 cgcacacaag ctgttgtcag tggccggaga agtagacccc gtgaaagaga ccgggaacga
     3061 gagcgagacc gccctagaga taacagacga gacagagagc gagatagagg acgtgataga
     3121 gaaagagaaa gagagcgatt atgtgatcga gacagagacc gaggggagag aggtcgatat
     3181 agaagataac tcgagcatgc atctagaggg ccctattcta tagtgtcacc taaatgctag
     3241 agctcgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc
     3301 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga
     3361 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca
     3421 ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc
     3481 tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg
     3541 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     3601 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     3661 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     3721 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     3781 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     3841 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     3901 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta
     3961 attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     4021 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     4081 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     4141 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     4201 tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag
     4261 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt
     4321 atatccattt tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa
     4381 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
     4441 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
     4501 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca
     4561 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
     4621 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
     4681 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     4741 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
     4801 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
     4861 ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc
     4921 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     4981 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     5041 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     5101 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     5161 tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag
     5221 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
     5281 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact
     5341 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     5401 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     5461 atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc
     5521 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     5581 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     5641 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     5701 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     5761 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     5821 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     5881 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     5941 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     6001 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     6061 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     6121 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     6181 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     6241 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     6301 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     6361 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     6421 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     6481 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     6541 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     6601 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     6661 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     6721 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     6781 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     6841 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     6901 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     6961 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     7021 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     7081 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     7141 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     7201 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     7261 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     7321 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     7381 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     7441 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     7501 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     7561 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     7621 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtc
//
