LOCUS       sequence-396937-        5967 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 5967)
  TITLE     Izaurralde lab plasmids
REFERENCE   2  (bases 1 to 5967)
  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-V5-SBP-C1-HsYTHDF3_AB
FEATURES             Location/Qualifiers
     source          1..5967
                     /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             668..709
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             731..844
                     /codon_start=1
                     /product="streptavidin-binding peptide"
                     /label=SBP
                     /note="selected from a peptide library; binds streptavidin 
                     with nanomolar affinity (Keefe et al., 2001)"
                     /translation="MDEKTTGWGGGHVVEGLAGELEQLRARLEHHPQGQREP"
     polyA_signal    2755..2876
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2792..2811)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2846..2865
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2883..3338)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3020..3041)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3232..3251
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3365..3469
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3471..3828
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3471..3662
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3494..3514)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3679..3814
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3863..4657
                     /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             3863..4657
                     /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(3917..3936)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4527..4546
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4845..4864)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4889..4936
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5265..5853
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5754..5773
                     /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 caccatgggt aagcctatcc ctaaccctct cctcggtctc gattctacgc ccggaagcgg
      721 tagcggtacg atggacgaga agaccaccgg ctgggggggc ggccacgtgg tggagggcct
      781 ggccggcgag ctggagcagc tgcgggccag gctggagcac caccctcagg gccagcggga
      841 gcccagatct cgagcaatgt cagccactag cgtggatcag agacctaaag ggcaaggaaa
      901 taaagtttca gtacaaaacg gttcgattca tcaaaaagat gctgtaaatg atgatgattt
      961 tgagccatac ttaagtagcc agacaaatca gagtaacagc tatccaccaa tgtcagatcc
     1021 atacatgcct agttactatg ctccatccat tggatttcca tattctcttg gggaagcagc
     1081 gtggtccaca gctggagacc agcctatgcc atatctgaca acctatggac aaatgagtaa
     1141 tggagaacat cactatatac cagatggtgt atttagtcaa cctggggcat taggaaatac
     1201 ccctccattt cttggtcaac atggatttaa cttttttcct ggtaatgctg atttctctac
     1261 atgggggaca agtggatctc agggacaatc aacacaaagt tctgcttata gtagcagtta
     1321 tggctatcca cctagttctc ttgggagagc tattactgat ggacaggctg gatttggcaa
     1381 tgatactttg agtaaggtgc ctggcattag cagtattgag caaggcatga ctggactgaa
     1441 aattggtggt gacctgacag ctgcagtgac aaaaactgta ggtacagctt tgagcagcag
     1501 tggtatgact agcattgcaa ccaatagtgt gcccccagtt agcagtgcag cacctaaacc
     1561 aacctcctgg gctgccattg ccagaaagcc tgccaaacct caaccgaaac ttaaacccaa
     1621 gggcaatgtg ggaattgggg gttctgctgt accaccacct cctataaaac acaacatgaa
     1681 tattggaact tgggatgaaa aagggtcagt ggtaaaggct ccaccaaccc aaccagttct
     1741 gcctcctcaa actataatcc agcagcctca gccattaatt caaccaccac cattggtgca
     1801 aagccaactg cctcaacagc agcctcaacc accacaacca cagcagcaac aaggacctca
     1861 gccacaggcc cagcctcacc aagtgcagcc tcaacagcag cagctgcaga atcgctgggt
     1921 agctcctcgt aacaggggag caggcttcaa ccagaacaat ggagcgggca gtgaaaactt
     1981 tggtttaggt gttgtacctg tcagtgcttc accttctagt gtagaagtgc atcccgtgct
     2041 ggaaaagcta aaggccataa acaactataa tcccaaagac tttgattgga atctgaagaa
     2101 tggacgtgtg tttataatta aaagctactc tgaggatgac atacatcgtt ccattaaata
     2161 ctctatctgg tgtagtactg agcatggtaa taagcgtttg gatgcagctt accgttccct
     2221 gaatgggaaa ggcccactct atttactctt cagtgtgaat ggcagtggac atttttgtgg
     2281 agtggctgaa atgaagtctg ttgtggacta taatgcgtat gctggtgtct ggtctcagga
     2341 taagtggaag ggcaaatttg aagttaaatg gatctttgtc aaagatgttc ccaataacca
     2401 attacggcat attcgcttag aaaataatga caacaaaccg gttaccaatt caagggacac
     2461 tcaagaggta cccctagaaa aagctaagca agtgcttaaa ataattgcta ctttcaagca
     2521 taccacctca atctttgatg actttgcaca ttatgaaaag cgtcaagaag aggaggaagc
     2581 catgcgtagg gagagaaata gaaacaaaca ataaccgcgg gcccgggatc caccggatct
     2641 agataactga tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac
     2701 ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg
     2761 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     2821 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaacgc
     2881 gtaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
     2941 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
     3001 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
     3061 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
     3121 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
     3181 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
     3241 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
     3301 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa
     3361 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     3421 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagtc ctgaggcgga
     3481 aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     3541 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     3601 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     3661 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     3721 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
     3781 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaga tcgatcaaga
     3841 gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc
     3901 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga
     3961 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
     4021 gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac
     4081 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
     4141 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt
     4201 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
     4261 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt
     4321 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag
     4381 gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt
     4441 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
     4501 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg
     4561 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg
     4621 catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg
     4681 accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat
     4741 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg
     4801 gatctcatgc tggagttctt cgcccaccct agggggaggc taactgaaac acggaaggag
     4861 acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggt
     4921 gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc
     4981 ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc
     5041 ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata
     5101 gcctcaggtt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     5161 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     5221 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     5281 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     5341 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     5401 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     5461 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     5521 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     5581 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     5641 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     5701 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     5761 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     5821 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     5881 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     5941 gtggataacc gtattaccgc catgcat
//
