LOCUS       sequence-228186-        6001 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6001)
  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 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"; journalName: "Proc Natl 
            Acad Sci U S A. 2009 Dec 22. 106(51):21591-6."
COMMENT     Sequence Label: pT7-EGFP-C1-HsDCP2
FEATURES             Location/Qualifiers
     source          1..6001
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        119..422
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        423..626
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     577..597
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     671..690
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        671..689
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      717..726
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(723..725,726..728,729..1439)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(723..725,726..728,729..1439)
                     /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(768..789)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1029..1048)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1376..1397
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2847..2968
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2884..2903)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2938..2957
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2975..3430)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3112..3133)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3324..3343
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3457..3561
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3563..3920
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3563..3754
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3586..3606)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3771..3906
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3955..4749
                     /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             3955..4749
                     /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(4009..4028)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4619..4638
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4937..4956)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4981..5028
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5357..5945
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5846..5865
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg ccatgcatta
       61 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
      121 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
      181 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
      241 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
      301 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
      361 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
      421 tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat
      481 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
      541 actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac
      601 ggtgggaggt ctatataagc agagctctct ggctaactag agaacccact gcttactggc
      661 ttatcgaaat taatacgact cactataggg agacccaagc tctagcgcta ccggtcgcca
      721 ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg
      781 acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct
      841 acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca
      901 ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga
      961 agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct
     1021 tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc
     1081 tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc
     1141 acaagctgga gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga
     1201 acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg
     1261 ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc
     1321 actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg
     1381 tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt
     1441 ccggactcag atctatggag accaaacggg tggagattcc cggcagcgtc ctggacgatc
     1501 tctgcagccg atttattttg catattccca gcgaggaaag agacaatgca atccgagtgt
     1561 gttttcagat tgaacttgcc cattggtttt acttggattt ctacatgcag aacacaccag
     1621 gattacctca gtgtgggata agagactttg ctaaagctgt cttcagtcat tgtccgtttt
     1681 tgctgcctca aggtgaagat gtggaaaaag ttttggatga atggaaggaa tataaaatgg
     1741 gagtaccaac atatggtgca attattcttg atgagacact tgaaaatgta ctactggttc
     1801 aggggtacct agcaaaatca ggctggggat ttccaaaagg aaaagtaaat aaagaagaag
     1861 ctcctcatga ttgtgctgct agagaggtct ttgaagaaac tggttttgat atcaaagact
     1921 atatttgtaa ggatgattac attgaacttc gaatcaatga ccagcttgct cgtttgtaca
     1981 tcattccagg aattccaaaa gacacaaaat ttaacccaaa aactagaaga gaaattcgga
     2041 acattgagtg gttctctatt gagaaattgc cttgtcatag aaatgatatg acccccaaat
     2101 ccaaacttgg tttggcacct aacaaatttt ttatggccat tccctttatc agaccattaa
     2161 gggactggct ttctcgaaga tttggcgatt cctcagacag tgacaatgga ttttcctcaa
     2221 ctggtagcac gccggctaaa cccactgtgg aaaaattgag tcgaaccaaa ttccgccaca
     2281 gtcagcagtt atttcctgac ggttctcctg gtgaccagtg ggtaaagcac aggcaaccac
     2341 tgcagcaaaa gccatataat aatcattctg aaatgtctga ccttttaaaa ggaaagaatc
     2401 aaagtatgag gggaaatggc agaaaacagt atcaagattc acctaatcaa aagaaaagaa
     2461 caaatgggct tcagccagca aagcagcaga attctttgat gaagtgtgaa aagaaacttc
     2521 atccacggaa acttcaggat aattttgaaa cagatgctgt atatgacttg cctagctcca
     2581 gtgaagacca gttgctagaa catgccgagg gacagcccgt ggcatgtaat ggacattgca
     2641 agttcccctt ttcatccaga gcctttttga gtttcaagtt tgaccataat gctataatga
     2701 aaatcttgga cctttgagga tccaccggat ctagataact gatcataatc agccatacca
     2761 catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac
     2821 ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat
     2881 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     2941 gtttgtccaa actcatcaat gtatcttaac gcgtaaattg taagcgttaa tattttgtta
     3001 aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc
     3061 aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg
     3121 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat
     3181 cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc
     3241 cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag
     3301 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg
     3361 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta
     3421 cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     3481 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     3541 taatattgaa aaaggaagag tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt
     3601 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     3661 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     3721 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     3781 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     3841 cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg
     3901 gaggcctagg cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt
     3961 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     4021 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     4081 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaagac
     4141 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     4201 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     4261 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     4321 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     4381 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     4441 caggggctcg cgccagccga actgttcgcc aggctcaagg cgagcatgcc cgacggcgag
     4501 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     4561 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     4621 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     4681 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     4741 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat
     4801 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     4861 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     4921 ctagggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga
     4981 cggcaataaa aagacagaat aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg
     5041 ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg
     5101 cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg
     5161 cagccaacgt cggggcggca ggccctgcca tagcctcagg ttactcatat atactttaga
     5221 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     5281 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     5341 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     5401 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     5461 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     5521 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     5581 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     5641 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     5701 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     5761 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     5821 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     5881 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     5941 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     6001 a
//
