LOCUS       Exported                7665 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Expression of FLAG-HA-tagged human YTHDC1.
ACCESSION   .
VERSION     .
KEYWORDS    pcDNA3-FLAG-HA-hYTHDC1
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7665)
  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 7665)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7665
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     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
                     /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..4303
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4154..4289
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4216..4235
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4370..5164
                     /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(4424..4443)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5034..5053
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5338..5459
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5375..5394)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5429..5448
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5508..5524)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5508..5524)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5521..5543)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5532..5548
                     /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(5556..5586)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5601..5622
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5739..5756)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5910..6498)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5990..6009)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6669..7529)
                     /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     7292..7311
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7530..7634)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct 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 atgggtggac 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 atcggggcat 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 ggggatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta
     3961 attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc aggcaggcag
     4021 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
     4081 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
     4141 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     4201 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca
     4261 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg
     4321 tatatccatt ttcggatctg atcaagagac aggatgagga tcgtttcgca tgattgaaca
     4381 agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg
     4441 ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg
     4501 cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc
     4561 agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt
     4621 cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc
     4681 atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca
     4741 tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc
     4801 acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg
     4861 gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct
     4921 cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc
     4981 tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc
     5041 tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta
     5101 cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt
     5161 ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga
     5221 gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac
     5281 gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac
     5341 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     5401 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
     5461 catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt
     5521 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     5581 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     5641 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     5701 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     5761 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     5821 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     5881 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     5941 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     6001 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     6061 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc acgctgtagg
     6121 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     6181 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     6241 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     6301 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt
     6361 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     6421 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     6481 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     6541 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     6601 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     6661 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     6721 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     6781 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     6841 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     6901 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     6961 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     7021 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     7081 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     7141 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     7201 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     7261 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     7321 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     7381 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     7441 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     7501 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     7561 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     7621 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtc
//
