LOCUS       sequence-474961-        6249 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsYTHDF3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6249)
  TITLE     Izaurralde lab plasmids
REFERENCE   2  (bases 1 to 6249)
  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"
COMMENT     Sequence Label: pT7-MS2-HA-C1-HsYTHDF3_AB
FEATURES             Location/Qualifiers
     source          1..6249
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     613..632
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        613..631
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      659..668
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1097..1123
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    3037..3158
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3074..3093)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3128..3147
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3165..3620)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3302..3323)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3514..3533
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3647..3751
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3753..4110
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3753..3944
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3776..3796)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3961..4096
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4145..4939
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             4145..4939
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4199..4218)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4809..4828
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5127..5146)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5171..5218
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5547..6135
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6036..6055
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca ctgcttactg
      601 gcttatcgaa attaatacga ctcactatag ggagacccaa gctctagcgc taccggtcgc
      661 caccatggct tctaacttta ctcagttcgt tctcgtcgac aatggcggaa ctggcgacgt
      721 gactgtcgcc ccaagcaact tcgctaacgg ggtcgctgaa tggatcagct ctaactcgcg
      781 atcacaggct tacaaagtaa cctgtagcgt tcgtcagagc tctgcgcaga atcgcaaata
      841 caccatcaaa gtcgaggtgc ctaaagtggc aacccagact gttggtggtg aagagcttcc
      901 tgtagccgga tggagatctt acttaaatat ggaactaacc attccaattt tcgccacgaa
      961 ttccgactgc gagcttattg ttaaggcaat gcaaggtctc ctaaaagatg gaaacccgat
     1021 tccctcggcc atcgcggcca actccggcat ctacggaggt ggaggtggat ctggtggatc
     1081 tcctccacat atggcttacc catacgatgt tccagattac gctcctccat ctcgagcaat
     1141 gtcagccact agcgtggatc agagacctaa agggcaagga aataaagttt cagtacaaaa
     1201 cggttcgatt catcaaaaag atgctgtaaa tgatgatgat tttgagccat acttaagtag
     1261 ccagacaaat cagagtaaca gctatccacc aatgtcagat ccatacatgc ctagttacta
     1321 tgctccatcc attggatttc catattctct tggggaagca gcgtggtcca cagctggaga
     1381 ccagcctatg ccatatctga caacctatgg acaaatgagt aatggagaac atcactatat
     1441 accagatggt gtatttagtc aacctggggc attaggaaat acccctccat ttcttggtca
     1501 acatggattt aacttttttc ctggtaatgc tgatttctct acatggggga caagtggatc
     1561 tcagggacaa tcaacacaaa gttctgctta tagtagcagt tatggctatc cacctagttc
     1621 tcttgggaga gctattactg atggacaggc tggatttggc aatgatactt tgagtaaggt
     1681 gcctggcatt agcagtattg agcaaggcat gactggactg aaaattggtg gtgacctgac
     1741 agctgcagtg acaaaaactg taggtacagc tttgagcagc agtggtatga ctagcattgc
     1801 aaccaatagt gtgcccccag ttagcagtgc agcacctaaa ccaacctcct gggctgccat
     1861 tgccagaaag cctgccaaac ctcaaccgaa acttaaaccc aagggcaatg tgggaattgg
     1921 gggttctgct gtaccaccac ctcctataaa acacaacatg aatattggaa cttgggatga
     1981 aaaagggtca gtggtaaagg ctccaccaac ccaaccagtt ctgcctcctc aaactataat
     2041 ccagcagcct cagccattaa ttcaaccacc accattggtg caaagccaac tgcctcaaca
     2101 gcagcctcaa ccaccacaac cacagcagca acaaggacct cagccacagg cccagcctca
     2161 ccaagtgcag cctcaacagc agcagctgca gaatcgctgg gtagctcctc gtaacagggg
     2221 agcaggcttc aaccagaaca atggagcggg cagtgaaaac tttggtttag gtgttgtacc
     2281 tgtcagtgct tcaccttcta gtgtagaagt gcatcccgtg ctggaaaagc taaaggccat
     2341 aaacaactat aatcccaaag actttgattg gaatctgaag aatggacgtg tgtttataat
     2401 taaaagctac tctgaggatg acatacatcg ttccattaaa tactctatct ggtgtagtac
     2461 tgagcatggt aataagcgtt tggatgcagc ttaccgttcc ctgaatggga aaggcccact
     2521 ctatttactc ttcagtgtga atggcagtgg acatttttgt ggagtggctg aaatgaagtc
     2581 tgttgtggac tataatgcgt atgctggtgt ctggtctcag gataagtgga agggcaaatt
     2641 tgaagttaaa tggatctttg tcaaagatgt tcccaataac caattacggc atattcgctt
     2701 agaaaataat gacaacaaac cggttaccaa ttcaagggac actcaagagg tacccctaga
     2761 aaaagctaag caagtgctta aaataattgc tactttcaag cataccacct caatctttga
     2821 tgactttgca cattatgaaa agcgtcaaga agaggaggaa gccatgcgta gggagagaaa
     2881 tagaaacaaa caataaccgc gggcccggga tccaccggat ctagataact gatcataatc
     2941 agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg
     3001 aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat
     3061 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     3121 tctagttgtg gtttgtccaa actcatcaat gtatcttaac gcgtaaattg taagcgttaa
     3181 tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc
     3241 cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt
     3301 tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa
     3361 aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg
     3421 gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg
     3481 acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc
     3541 tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa
     3601 tgcgccgcta cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     3661 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     3721 aatgcttcaa taatattgaa aaaggaagag tcctgaggcg gaaagaacca gctgtggaat
     3781 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     3841 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     3901 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     3961 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     4021 tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga
     4081 ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt
     4141 tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct
     4201 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct
     4261 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga
     4321 actgcaagac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc
     4381 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg
     4441 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc
     4501 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca
     4561 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga
     4621 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgagcatgcc
     4681 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga
     4741 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca
     4801 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg
     4861 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct
     4921 tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc
     4981 aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
     5041 atcgttttcc gggacaccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
     5101 ttcgcccacc ctagggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc
     5161 cgcgctatga cggcaataaa aagacagaat aaaacgcacg gtgttgggtc gtttgttcat
     5221 aaacgcgggg ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg
     5281 ggccaatacg cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc
     5341 ccagggctcg cagccaacgt cggggcggca ggccctgcca tagcctcagg ttactcatat
     5401 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     5461 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     5521 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     5581 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     5641 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     5701 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     5761 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     5821 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     5881 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     5941 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     6001 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     6061 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     6121 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     6181 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     6241 gccatgcat
//
