LOCUS       sequence-389938-        6513 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 6513)
  TITLE     Izaurralde lab plasmids
REFERENCE   2  (bases 1 to 6513)
  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-EGFP-C1-HsYTHDF3_AB
FEATURES             Location/Qualifiers
     source          1..6513
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     553..572
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        553..571
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      599..608
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(605..607,608..610,611..1321)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(605..607,608..610,611..1321)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(650..671)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(911..930)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1258..1279
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3241..3362
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3278..3297)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3332..3351
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3369..3824)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3506..3527)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3718..3737
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3851..3955
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3957..4314
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3957..4148
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3980..4000)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4165..4300
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4349..5143
                     /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             4349..5143
                     /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(4403..4422)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5013..5032
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5331..5350)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5375..5422
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5751..6339
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6240..6259
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca ctgcttactg
      541 gcttatcgaa attaatacga ctcactatag ggagacccaa gctctagcgc taccggtcgc
      601 caccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct
      661 ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac
      721 ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc
      781 caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat
      841 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat
      901 cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac
      961 cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg
     1021 gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa
     1081 gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct
     1141 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa
     1201 ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat
     1261 ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa
     1321 gtccggactc agatctcgag caatgtcagc cactagcgtg gatcagagac ctaaagggca
     1381 aggaaataaa gtttcagtac aaaacggttc gattcatcaa aaagatgctg taaatgatga
     1441 tgattttgag ccatacttaa gtagccagac aaatcagagt aacagctatc caccaatgtc
     1501 agatccatac atgcctagtt actatgctcc atccattgga tttccatatt ctcttgggga
     1561 agcagcgtgg tccacagctg gagaccagcc tatgccatat ctgacaacct atggacaaat
     1621 gagtaatgga gaacatcact atataccaga tggtgtattt agtcaacctg gggcattagg
     1681 aaatacccct ccatttcttg gtcaacatgg atttaacttt tttcctggta atgctgattt
     1741 ctctacatgg gggacaagtg gatctcaggg acaatcaaca caaagttctg cttatagtag
     1801 cagttatggc tatccaccta gttctcttgg gagagctatt actgatggac aggctggatt
     1861 tggcaatgat actttgagta aggtgcctgg cattagcagt attgagcaag gcatgactgg
     1921 actgaaaatt ggtggtgacc tgacagctgc agtgacaaaa actgtaggta cagctttgag
     1981 cagcagtggt atgactagca ttgcaaccaa tagtgtgccc ccagttagca gtgcagcacc
     2041 taaaccaacc tcctgggctg ccattgccag aaagcctgcc aaacctcaac cgaaacttaa
     2101 acccaagggc aatgtgggaa ttgggggttc tgctgtacca ccacctccta taaaacacaa
     2161 catgaatatt ggaacttggg atgaaaaagg gtcagtggta aaggctccac caacccaacc
     2221 agttctgcct cctcaaacta taatccagca gcctcagcca ttaattcaac caccaccatt
     2281 ggtgcaaagc caactgcctc aacagcagcc tcaaccacca caaccacagc agcaacaagg
     2341 acctcagcca caggcccagc ctcaccaagt gcagcctcaa cagcagcagc tgcagaatcg
     2401 ctgggtagct cctcgtaaca ggggagcagg cttcaaccag aacaatggag cgggcagtga
     2461 aaactttggt ttaggtgttg tacctgtcag tgcttcacct tctagtgtag aagtgcatcc
     2521 cgtgctggaa aagctaaagg ccataaacaa ctataatccc aaagactttg attggaatct
     2581 gaagaatgga cgtgtgttta taattaaaag ctactctgag gatgacatac atcgttccat
     2641 taaatactct atctggtgta gtactgagca tggtaataag cgtttggatg cagcttaccg
     2701 ttccctgaat gggaaaggcc cactctattt actcttcagt gtgaatggca gtggacattt
     2761 ttgtggagtg gctgaaatga agtctgttgt ggactataat gcgtatgctg gtgtctggtc
     2821 tcaggataag tggaagggca aatttgaagt taaatggatc tttgtcaaag atgttcccaa
     2881 taaccaatta cggcatattc gcttagaaaa taatgacaac aaaccggtta ccaattcaag
     2941 ggacactcaa gaggtacccc tagaaaaagc taagcaagtg cttaaaataa ttgctacttt
     3001 caagcatacc acctcaatct ttgatgactt tgcacattat gaaaagcgtc aagaagagga
     3061 ggaagccatg cgtagggaga gaaatagaaa caaacaataa ccgcgggccc gggatccacc
     3121 ggatctagat aactgatcat aatcagccat accacatttg tagaggtttt acttgcttta
     3181 aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt
     3241 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     3301 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     3361 taacgcgtaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat
     3421 cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata
     3481 gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt
     3541 ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc
     3601 atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa
     3661 agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg
     3721 gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt
     3781 aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg
     3841 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     3901 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtcctga
     3961 ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     4021 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     4081 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     4141 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     4201 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct
     4261 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaagatcga
     4321 tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc
     4381 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg
     4441 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac
     4501 cgacctgtcc ggtgccctga atgaactgca agacgaggca gcgcggctat cgtggctggc
     4561 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg
     4621 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga
     4681 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg
     4741 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg
     4801 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt
     4861 cgccaggctc aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc
     4921 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg
     4981 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga
     5041 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc
     5101 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc
     5161 gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc
     5221 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag
     5281 cgcggggatc tcatgctgga gttcttcgcc caccctaggg ggaggctaac tgaaacacgg
     5341 aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg
     5401 cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc
     5461 gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc
     5521 caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct
     5581 gccatagcct caggttactc atatatactt tagattgatt taaaacttca tttttaattt
     5641 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     5701 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     5761 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     5821 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     5881 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     5941 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     6001 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     6061 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     6121 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     6181 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     6241 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     6301 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     6361 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     6421 gattctgtgg ataaccgtat taccgccatg cattagttat taatagtaat caattacggg
     6481 gtcattagtt catagcccat atatggagtt ccg
//
