LOCUS       sequence-288952-        6527 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsDCP1a-withDmTD.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6527)
  AUTHORS   Tritschler F, Braun JE, Motz C, Igreja C, Haas G, Truffault V, 
            Izaurralde E, Weichenrieder O
  TITLE     DCP1 forms asymmetric trimers to assemble into active mRNA decapping
            complexes in metazoa.
  JOURNAL   Proc Natl Acad Sci U S A. 2009 Dec 22. 106(51):21591-6.
  PUBMED    19966221
REFERENCE   2  (bases 1 to 6527)
  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"; journalName: "Proc Natl 
            Acad Sci U S A. 2009 Dec 22. 106(51):21591-6."
COMMENT     Sequence Label: pT7-EGFP-C1-HsDCP1a-withDmTD_H
FEATURES             Location/Qualifiers
     source          1..6527
                     /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    3263..3384
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3300..3319)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3354..3373
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3391..3846)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3528..3549)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3740..3759
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3873..3977
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3979..4336
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3979..4170
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4002..4022)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4187..4322
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4371..5165
                     /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             4371..5165
                     /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(4425..4444)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5035..5054
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5353..5372)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5397..5444
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5773..6361
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6262..6281
                     /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 atcaatggag gcgctgagtc gagctgggca ggagatgagc
     1381 ctagcggccc tgaagcaaca cgacccctat atcaccagca tcgcagacct cacgggccag
     1441 gtcgctctgt acaccttctg ccccaaggcc aaccagtggg agaagactga tatagaaggg
     1501 accttattcg tatatcgaag gtcagcttcc ccttaccatg gttttaccat tgtgaatcga
     1561 ctaaatatgc acaatctagt cgaaccagtg aataaagatt tggaatttca gctccatgaa
     1621 ccatttcttc tgtatagaaa tgcaagcttg tcgatatata gtatctggtt ttatgacaag
     1681 aatgactgtc accgcatagc aaaactcatg gctgatgtgg tagaagagga gacacggcga
     1741 tcccagcaag ctgctcggga caaacagagt cccagccagg ccaatggctg cagcgaccac
     1801 aggcccatcg acatcctgga gatgctgagc agagccaagg atgagtatga gaggaatcag
     1861 atgggtgact caaatatctc cagccctggg ttacagccaa gcactcagct ctccaatctg
     1921 ggaagcaccg agactctaga agaaatgccc tccgggtcac aggataagtc tgctccatct
     1981 ggacacaagc atctgacggt agaagagtta tttggaacct ctttgccaaa ggaacaacca
     2041 gcagttgtgg gtctggattc agaagaaatg gagaggttgc caggagatgc ctcccagaaa
     2101 gagcccaatt cattcctacc atttcccttt gagcagttag gaggagcccc tcaatcagaa
     2161 accctgggtg tcccttctgc tgcccaccat tcagtccagc ctgaaatcac caccccggtg
     2221 ctaatcactc cagcctccat cacacagtcc aatgaaaagc atgctccaac ctacacaatc
     2281 ccgttgagcc ctgttctcag tcccactctg ccagctgaag ctcctactgc acaggttccc
     2341 cccagcttac ctcgaaacag caccatgatg caggcagtga agaccacgcc tagacagagg
     2401 tctccactcc tgaaccagcc agtccctgag ctaagccatg ccagtctgat tgccaaccag
     2461 agccccttca gggccccatt gaacgtgacg aacacagctg gcacatccct cccaagcgtt
     2521 gatcttctcc agaaactcag gttgacccca cagcatgacc aaatacagac acaaccactt
     2581 gggaaaggtg caatggtagc cagcttttct ccggcagctg gtcagctagc cacacctgag
     2641 agcttcatag agcctccctc taagacagca gcagcaagag tggcggcctc agcctccctg
     2701 agcaacatgg tgcttgctcc ccttcagtct atgcagcaga accaggatcc tgaagtattt
     2761 gtgcagccta aggtgttatc cagtgccatc ccggttgcag gcgccccact ggttactgca
     2821 acgaccactg cagtgtcttc agtcctgctg gccccaagtg ttttccagca gacagttaca
     2881 agatcttcgg accttgagag gaaagccagc tccccttctc ctctaactat tggaacgcca
     2941 gaaagtcagc agccacagca gccactgctg aactcgacgc agttcgtcca ggccttcacg
     3001 taccttatcc agaacgataa ggagtttgcc aacaagctgc acaaggccta cctgaacggc
     3061 tgttccaatc tactgctgga ctccagctcc acatatcaat gagaattctg cagtcgacgg
     3121 taccgcgggc ccgggatcca ccggatctag ataactgatc ataatcagcc ataccacatt
     3181 tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa
     3241 aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag
     3301 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     3361 gtccaaactc atcaatgtat cttaacgcgt aaattgtaag cgttaatatt ttgttaaaat
     3421 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa
     3481 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca
     3541 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
     3601 gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta
     3661 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg
     3721 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa
     3781 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
     3841 gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     3901 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     3961 attgaaaaag gaagagtcct gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg
     4021 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     4081 gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     4141 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac
     4201 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     4261 ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     4321 cctaggcttt tgcaaagatc gatcaagaga caggatgagg atcgtttcgc atgattgaac
     4381 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
     4441 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
     4501 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caagacgagg
     4561 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg
     4621 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt
     4681 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc
     4741 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag
     4801 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg
     4861 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc
     4921 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt
     4981 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg
     5041 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt
     5101 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct
     5161 tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg
     5221 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
     5281 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccctag
     5341 ggggaggcta actgaaacac ggaaggagac aataccggaa ggaacccgcg ctatgacggc
     5401 aataaaaaga cagaataaaa cgcacggtgt tgggtcgttt gttcataaac gcggggttcg
     5461 gtcccagggc tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg
     5521 cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc
     5581 caacgtcggg gcggcaggcc ctgccatagc ctcaggttac tcatatatac tttagattga
     5641 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     5701 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     5761 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     5821 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     5881 ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt
     5941 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     6001 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     6061 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     6121 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     6181 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     6241 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     6301 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     6361 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     6421 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcca tgcattagtt
     6481 attaatagta atcaattacg gggtcattag ttcatagccc atatatg
//
