LOCUS       sequence-290247-        6518 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsDCP1a-N74G.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6518)
  AUTHORS   Chang CT, Bercovich N, Loh B, Jonas S, Izaurralde E
  TITLE     The activation of the decapping enzyme DCP2 by DCP1 occurs on the 
            EDC4 scaffold and involves a conserved loop in DCP1.
  JOURNAL   Nucleic Acids Res. 2014 Apr;42(8):5217-33. doi: 10.1093/nar/gku129. 
            Epub 2014 Feb 8.
  PUBMED    24510189
REFERENCE   2  (bases 1 to 6518)
  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: 
            "10.1093/nar/gku129"; journalName: "Nucleic Acids Res"; date: 
            "2014-04"; volume: "42"; issue: "8"; pages: "5217-33"
COMMENT     Sequence Label: pT7-EGFP-C1-HsDCP1a-N74G_V
FEATURES             Location/Qualifiers
     source          1..6518
                     /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             join(665..667,668..670,671..1381)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(665..667,668..670,671..1381)
                     /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(710..731)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(971..990)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1318..1339
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3306..3427
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3343..3362)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3397..3416
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3434..3889)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3571..3592)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3783..3802
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3916..4020
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4022..4379
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4022..4213
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4045..4065)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4230..4365
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4414..5208
                     /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             4414..5208
                     /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(4468..4487)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5078..5097
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5396..5415)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5440..5487
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5816..6404
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6305..6324
                     /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 caccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct
      721 ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac
      781 ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc
      841 caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat
      901 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat
      961 cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac
     1021 cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg
     1081 gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa
     1141 gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct
     1201 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa
     1261 ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat
     1321 ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa
     1381 gtccggactc agatcaatgg aggcgctgag tcgagctggg caggagatga gcctagcggc
     1441 cctgaagcaa cacgacccct atatcaccag catcgcagac ctcacgggcc aggtcgctct
     1501 gtacaccttc tgccccaagg ccaaccagtg ggagaagact gatatagaag ggaccttatt
     1561 cgtatatcga aggtcagctt ccccttacca tggttttacc attgtgaatc gactaggtat
     1621 gcacaatcta gtcgaaccag tgaataaaga tttggaattt cagctccatg aaccatttct
     1681 tctgtataga aatgcaagct tgtcgatata tagtatctgg ttttatgaca agaatgactg
     1741 tcaccgcata gcaaaactca tggctgatgt ggtagaagag gagacacggc gatcccagca
     1801 agctgctcgg gacaaacaga gtcccagcca ggccaatggc tgcagcgacc acaggcccat
     1861 cgacatcctg gagatgctga gcagagccaa ggatgagtat gagaggaatc agatgggtga
     1921 ctcaaatatc tccagccctg ggttacagcc aagcactcag ctctccaatc tgggaagcac
     1981 cgagactcta gaagaaatgc cctccgggtc acaggataag tctgctccat ctggacacaa
     2041 gcatctgacg gtagaagagt tatttggaac ctctttgcca aaggaacaac cagcagttgt
     2101 gggtctggat tcagaagaaa tggagaggtt gccaggagat gcctcccaga aagagcccaa
     2161 ttcattccta ccatttccct ttgagcagtt aggaggagcc cctcaatcag aaaccctggg
     2221 tgtcccttct gctgcccacc attcagtcca gcctgaaatc accaccccgg tgctaatcac
     2281 tccagcctcc atcacacagt ccaatgaaaa gcatgctcca acctacacaa tcccgttgag
     2341 ccctgttctc agtcccactc tgccagctga agctcctact gcacaggttc cccccagctt
     2401 acctcgaaac agcaccatga tgcaggcagt gaagaccacg cctagacaga ggtctccact
     2461 cctgaaccag ccagtccctg agctaagcca tgccagtctg attgccaacc agagcccctt
     2521 cagggcccca ttgaacgtga cgaacacagc tggcacatcc ctcccaagcg ttgatcttct
     2581 ccagaaactc aggttgaccc cacagcatga ccaaatacag acacaaccac ttgggaaagg
     2641 tgcaatggta gccagctttt ctccggcagc tggtcagcta gccacacctg agagcttcat
     2701 agagcctccc tctaagacag cagcagcaag agtggcggcc tcagcctccc tgagcaacat
     2761 ggtgcttgct ccccttcagt ctatgcagca gaaccaggat cctgaagtat ttgtgcagcc
     2821 taaggtgtta tccagtgcca tcccggttgc aggcgcccca ctggttactg caacgaccac
     2881 tgcagtgtct tcagtcctgc tggccccaag tgttttccag cagacagtta caagatcttc
     2941 ggaccttgag aggaaagcca gctccccttc tcctctaact attggaacgc cagaaagtca
     3001 gagaaagcct tccattattc tcagcaagtc tcagctccag gatacattaa tacatctaat
     3061 aaagaatgat tccagcttcc tcagtacact tcatgaagtc tacttgcagg tgctgaccaa
     3121 gaacaaagac aaccacaacc tatgagaatt ctgcagtcga cggtaccgcg ggcccgggat
     3181 ccaccggatc tagataactg atcataatca gccataccac atttgtagag gttttacttg
     3241 ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg
     3301 ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     3361 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     3421 tatcttaacg cgtaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt
     3481 taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa
     3541 gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag
     3601 aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt
     3661 gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac
     3721 cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag
     3781 gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg
     3841 cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc aggtggcact
     3901 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     3961 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     4021 cctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
     4081 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     4141 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     4201 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     4261 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg
     4321 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaag
     4381 atcgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca
     4441 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
     4501 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
     4561 aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg
     4621 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
     4681 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
     4741 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
     4801 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
     4861 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
     4921 ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt gacccatggc
     4981 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
     5041 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
     5101 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
     5161 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg
     5221 ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg
     5281 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc
     5341 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc tagggggagg ctaactgaaa
     5401 cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa agacagaata
     5461 aaacgcacgg tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact
     5521 ctgtcgatac cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc
     5581 caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag
     5641 gccctgccat agcctcaggt tactcatata tactttagat tgatttaaaa cttcattttt
     5701 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     5761 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     5821 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     5881 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     5941 gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga
     6001 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     6061 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     6121 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     6181 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     6241 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     6301 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     6361 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     6421 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     6481 cccctgattc tgtggataac cgtattaccg ccatgcat
//
