LOCUS       sequence-405437-        6001 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsDCP2-E148Q.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6001)
  AUTHORS   Loh B, Jonas S, Izaurralde E
  TITLE     The SMG5-SMG7 heterodimer directly recruits the CCR4-NOT deadenylase
            complex to mRNAs containing nonsense codons via interaction with 
            POP2.
  JOURNAL   Genes Dev. 2013 Oct 1;27(19):2125-38. doi: 10.1101/gad.226951.113.
  PUBMED    24115769
REFERENCE   2  (bases 1 to 6001)
  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: 
            "10.1101/gad.226951"; journalName: "Genes Dev"; date: "2013-10-1- 
            1"; volume: "27"; issue: "19"; pages: "2125-38"
COMMENT     Sequence Label: pT7-EGFP-C1-HsDCP2-E148Q_U
FEATURES             Location/Qualifiers
     source          1..6001
                     /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    2729..2850
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2766..2785)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2820..2839
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2857..3312)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2994..3015)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3206..3225
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3339..3443
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3445..3802
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3445..3636
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3468..3488)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3653..3788
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3837..4631
                     /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             3837..4631
                     /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(3891..3910)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4501..4520
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4819..4838)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4863..4910
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5239..5827
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5728..5747
                     /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 agatctatgg agaccaaacg ggtggagatt cccggcagcg tcctggacga
     1381 tctctgcagc cgatttattt tgcatattcc cagcgaggaa agagacaatg caatccgagt
     1441 gtgttttcag attgaacttg cccattggtt ttacttggat ttctacatgc agaacacacc
     1501 aggattacct cagtgtggga taagagactt tgctaaagct gtcttcagtc attgtccgtt
     1561 tttgctgcct caaggtgaag atgtggaaaa agttttggat gaatggaagg aatataaaat
     1621 gggagtacca acatatggtg caattattct tgatgagaca cttgaaaatg tactactggt
     1681 tcaggggtac ctagcaaaat caggctgggg atttccaaaa ggaaaagtaa ataaagaaga
     1741 agctcctcat gattgtgctg ctagagaggt ctttgaacag actggttttg atatcaaaga
     1801 ctatatttgt aaggatgatt acattgaact tcgaatcaat gaccagcttg ctcgtttgta
     1861 catcattcca ggaattccaa aagacacaaa atttaaccca aaaactagaa gagaaattcg
     1921 gaacattgag tggttctcta ttgagaaatt gccttgtcat agaaatgata tgacccccaa
     1981 atccaaactt ggtttggcac ctaacaaatt ttttatggcc attcccttta tcagaccatt
     2041 aagggactgg ctttctcgaa gatttggcga ttcctcagac agtgacaatg gattttcctc
     2101 aactggtagc acgccggcta aacccactgt ggaaaaattg agtcgaacca aattccgcca
     2161 cagtcagcag ttatttcctg acggttctcc tggtgaccag tgggtaaagc acaggcaacc
     2221 actgcagcaa aagccatata ataatcattc tgaaatgtct gaccttttaa aaggaaagaa
     2281 tcaaagtatg aggggaaatg gcagaaaaca gtatcaagat tcacctaatc aaaagaaaag
     2341 aacaaatggg cttcagccag caaagcagca gaattctttg atgaagtgtg aaaagaaact
     2401 tcatccacgg aaacttcagg ataattttga aacagatgct gtatatgact tgcctagctc
     2461 cagtgaagac cagttgctag aacatgccga gggacagccc gtggcatgta atggacattg
     2521 caagttcccc ttttcatcca gagccttttt gagtttcaag tttgaccata atgctataat
     2581 gaaaatcttg gacctttgag gatccaccgg atctagataa ctgatcataa tcagccatac
     2641 cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa
     2701 acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa
     2761 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     2821 tggtttgtcc aaactcatca atgtatctta acgcgtaaat tgtaagcgtt aatattttgt
     2881 taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg
     2941 gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt
     3001 ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct
     3061 atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt
     3121 gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa
     3181 agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc
     3241 tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc
     3301 tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     3361 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     3421 aataatattg aaaaaggaag agtcctgagg cggaaagaac cagctgtgga atgtgtgtca
     3481 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     3541 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     3601 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     3661 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     3721 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt
     3781 tggaggccta ggcttttgca aagatcgatc aagagacagg atgaggatcg tttcgcatga
     3841 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     3901 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     3961 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcaag
     4021 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     4081 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     4141 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     4201 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     4261 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     4321 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg
     4381 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     4441 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     4501 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     4561 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     4621 agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc
     4681 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     4741 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     4801 ccctaggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat
     4861 gacggcaata aaaagacaga ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg
     4921 ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata
     4981 cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct
     5041 cgcagccaac gtcggggcgg caggccctgc catagcctca ggttactcat atatacttta
     5101 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     5161 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     5221 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     5281 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     5341 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     5401 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     5461 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     5521 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     5581 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     5641 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     5701 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     5761 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     5821 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     5881 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgccatgca
     5941 ttagttatta atagtaatca attacggggt cattagttca tagcccatat atggagttcc
     6001 g
//
