LOCUS       sequence-290261-        6001 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsDCP2-G96PV97P.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6001)
  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 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: 
            "10.1093/nar/gku129"; journalName: "Nucleic Acids Res"; date: 
            "2014-04"; volume: "42"; issue: "8"; pages: "5217-33"
COMMENT     Sequence Label: pT7-EGFP-C1-HsDCP2-G96PV97P_V
FEATURES             Location/Qualifiers
     source          1..6001
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        9..312
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        313..516
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     467..487
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     561..580
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        561..579
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      607..616
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(613..615,616..618,619..1329)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(613..615,616..618,619..1329)
                     /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(658..679)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(919..938)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1266..1287
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2737..2858
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2774..2793)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2828..2847
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2865..3320)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3002..3023)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3214..3233
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3347..3451
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3453..3810
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3453..3644
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3476..3496)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3661..3796
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3845..4639
                     /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             3845..4639
                     /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(3899..3918)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4509..4528
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4827..4846)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4871..4918
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5247..5835
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5736..5755
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
       61 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat
      121 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
      181 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
      241 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc
      301 gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac
      361 tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa
      421 aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt
      481 aggcgtgtac ggtgggaggt ctatataagc agagctctct ggctaactag agaacccact
      541 gcttactggc ttatcgaaat taatacgact cactataggg agacccaagc tctagcgcta
      601 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
      661 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
      721 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
      781 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
      841 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
      901 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
      961 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     1021 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     1081 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     1141 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1201 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
     1261 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     1321 ctgtacaagt ccggactcag atctatggag accaaacggg tggagattcc cggcagcgtc
     1381 ctggacgatc tctgcagccg atttattttg catattccca gcgaggaaag agacaatgca
     1441 atccgagtgt gttttcagat tgaacttgcc cattggtttt acttggattt ctacatgcag
     1501 aacacaccag gattacctca gtgtgggata agagactttg ctaaagctgt cttcagtcat
     1561 tgtccgtttt tgctgcctca aggtgaagat gtggaaaaag ttttggatga atggaaggaa
     1621 tataaaatgc caccaccaac atatggtgca attattcttg atgagacact tgaaaatgta
     1681 ctactggttc aggggtacct agcaaaatca ggctggggat ttccaaaagg aaaagtaaat
     1741 aaagaagaag ctcctcatga ttgtgctgct agagaggtct ttgaagaaac tggttttgat
     1801 atcaaagact atatttgtaa ggatgattac attgaacttc gaatcaatga ccagcttgct
     1861 cgtttgtaca tcattccagg aattccaaaa gacacaaaat ttaacccaaa aactagaaga
     1921 gaaattcgga acattgagtg gttctctatt gagaaattgc cttgtcatag aaatgatatg
     1981 acccccaaat ccaaacttgg tttggcacct aacaaatttt ttatggccat tccctttatc
     2041 agaccattaa gggactggct ttctcgaaga tttggcgatt cctcagacag tgacaatgga
     2101 ttttcctcaa ctggtagcac gccggctaaa cccactgtgg aaaaattgag tcgaaccaaa
     2161 ttccgccaca gtcagcagtt atttcctgac ggttctcctg gtgaccagtg ggtaaagcac
     2221 aggcaaccac tgcagcaaaa gccatataat aatcattctg aaatgtctga ccttttaaaa
     2281 ggaaagaatc aaagtatgag gggaaatggc agaaaacagt atcaagattc acctaatcaa
     2341 aagaaaagaa caaatgggct tcagccagca aagcagcaga attctttgat gaagtgtgaa
     2401 aagaaacttc atccacggaa acttcaggat aattttgaaa cagatgctgt atatgacttg
     2461 cctagctcca gtgaagacca gttgctagaa catgccgagg gacagcccgt ggcatgtaat
     2521 ggacattgca agttcccctt ttcatccaga gcctttttga gtttcaagtt tgaccataat
     2581 gctataatga aaatcttgga cctttgagga tccaccggat ctagataact gatcataatc
     2641 agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg
     2701 aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat
     2761 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     2821 tctagttgtg gtttgtccaa actcatcaat gtatcttaac gcgtaaattg taagcgttaa
     2881 tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc
     2941 cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt
     3001 tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa
     3061 aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg
     3121 gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg
     3181 acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc
     3241 tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa
     3301 tgcgccgcta cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     3361 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     3421 aatgcttcaa taatattgaa aaaggaagag tcctgaggcg gaaagaacca gctgtggaat
     3481 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     3541 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     3601 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     3661 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     3721 tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga
     3781 ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt
     3841 tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct
     3901 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct
     3961 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga
     4021 actgcaagac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc
     4081 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg
     4141 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc
     4201 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca
     4261 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga
     4321 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgagcatgcc
     4381 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga
     4441 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca
     4501 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg
     4561 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct
     4621 tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc
     4681 aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
     4741 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
     4801 ttcgcccacc ctagggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc
     4861 cgcgctatga cggcaataaa aagacagaat aaaacgcacg gtgttgggtc gtttgttcat
     4921 aaacgcgggg ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg
     4981 ggccaatacg cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc
     5041 ccagggctcg cagccaacgt cggggcggca ggccctgcca tagcctcagg ttactcatat
     5101 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     5161 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     5221 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     5281 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     5341 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta
     5401 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     5461 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     5521 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     5581 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     5641 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     5701 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     5761 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     5821 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     5881 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     5941 gccatgcatt agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat
     6001 g
//
