LOCUS       sequence-386571-        5917 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsDCP2_1-395.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5917)
  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 5917)
  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-HsDCP2_1-395_V
FEATURES             Location/Qualifiers
     source          1..5917
                     /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    2645..2766
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2682..2701)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2736..2755
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2773..3228)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2910..2931)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3122..3141
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3255..3359
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3361..3718
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3361..3552
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3384..3404)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3569..3704
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3753..4547
                     /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             3753..4547
                     /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(3807..3826)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4417..4436
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4735..4754)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4779..4826
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5155..5743
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5644..5663
                     /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 ctttgaagaa 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 attgaggatc
     2521 caccggatct agataactga tcataatcag ccataccaca tttgtagagg ttttacttgc
     2581 tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt
     2641 tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     2701 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     2761 atcttaacgc gtaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt
     2821 aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag
     2881 aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga
     2941 acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg
     3001 aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc
     3061 ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg
     3121 aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc
     3181 gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt
     3241 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     3301 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagtc
     3361 ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
     3421 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
     3481 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     3541 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     3601 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc
     3661 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaga
     3721 tcgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag
     3781 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
     3841 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
     3901 agaccgacct gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc
     3961 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
     4021 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
     4081 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
     4141 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
     4201 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
     4261 tgttcgccag gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg acccatggcg
     4321 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
     4381 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
     4441 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
     4501 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg
     4561 gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc
     4621 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
     4681 ccagcgcggg gatctcatgc tggagttctt cgcccaccct agggggaggc taactgaaac
     4741 acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa
     4801 aacgcacggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc
     4861 tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc
     4921 accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg
     4981 ccctgccata gcctcaggtt actcatatat actttagatt gatttaaaac ttcattttta
     5041 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     5101 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     5161 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     5221 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     5281 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     5341 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     5401 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     5461 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     5521 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     5581 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     5641 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     5701 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     5761 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     5821 ccctgattct gtggataacc gtattaccgc catgcattag ttattaatag taatcaatta
     5881 cggggtcatt agttcatagc ccatatatgg agttccg
//
