LOCUS       sequence-228181-        6518 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 6518)
  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 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"; journalName: "Proc Natl 
            Acad Sci U S A. 2009 Dec 22. 106(51):21591-6."
COMMENT     Sequence Label: pT7-EGFP-C1-HsDCP1a
FEATURES             Location/Qualifiers
     source          1..6518
                     /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    3364..3485
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3401..3420)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3455..3474
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3492..3947)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3629..3650)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3841..3860
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3974..4078
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4080..4437
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4080..4271
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4103..4123)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4288..4423
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4472..5266
                     /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             4472..5266
                     /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(4526..4545)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5136..5155
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5454..5473)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5498..5545
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5874..6462
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6363..6382
                     /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 atcaatggag gcgctgagtc gagctgggca ggagatgagc ctagcggccc
     1501 tgaagcaaca cgacccctat atcaccagca tcgcagacct cacgggccag gtcgctctgt
     1561 acaccttctg ccccaaggcc aaccagtggg agaagactga tatagaaggg accttattcg
     1621 tatatcgaag gtcagcttcc ccttaccatg gttttaccat tgtgaatcga ctaaatatgc
     1681 acaatctagt cgaaccagtg aataaagatt tggaatttca gctccatgaa ccatttcttc
     1741 tgtatagaaa tgcaagcttg tcgatatata gtatctggtt ttatgacaag aatgactgtc
     1801 accgcatagc aaaactcatg gctgatgtgg tagaagagga gacacggcga tcccagcaag
     1861 ctgctcggga caaacagagt cccagccagg ccaatggctg cagcgaccac aggcccatcg
     1921 acatcctgga gatgctgagc agagccaagg atgagtatga gaggaatcag atgggtgact
     1981 caaatatctc cagccctggg ttacagccaa gcactcagct ctccaatctg ggaagcaccg
     2041 agactctaga agaaatgccc tccgggtcac aggataagtc tgctccatct ggacacaagc
     2101 atctgacggt agaagagtta tttggaacct ctttgccaaa ggaacaacca gcagttgtgg
     2161 gtctggattc agaagaaatg gagaggttgc caggagatgc ctcccagaaa gagcccaatt
     2221 cattcctacc atttcccttt gagcagttag gaggagcccc tcaatcagaa accctgggtg
     2281 tcccttctgc tgcccaccat tcagtccagc ctgaaatcac caccccggtg ctaatcactc
     2341 cagcctccat cacacagtcc aatgaaaagc atgctccaac ctacacaatc ccgttgagcc
     2401 ctgttctcag tcccactctg ccagctgaag ctcctactgc acaggttccc cccagcttac
     2461 ctcgaaacag caccatgatg caggcagtga agaccacgcc tagacagagg tctccactcc
     2521 tgaaccagcc agtccctgag ctaagccatg ccagtctgat tgccaaccag agccccttca
     2581 gggccccatt gaacgtgacg aacacagctg gcacatccct cccaagcgtt gatcttctcc
     2641 agaaactcag gttgacccca cagcatgacc aaatacagac acaaccactt gggaaaggtg
     2701 caatggtagc cagcttttct ccggcagctg gtcagctagc cacacctgag agcttcatag
     2761 agcctccctc taagacagca gcagcaagag tggcggcctc agcctccctg agcaacatgg
     2821 tgcttgctcc ccttcagtct atgcagcaga accaggatcc tgaagtattt gtgcagccta
     2881 aggtgttatc cagtgccatc ccggttgcag gcgccccact ggttactgca acgaccactg
     2941 cagtgtcttc agtcctgctg gccccaagtg ttttccagca gacagttaca agatcttcgg
     3001 accttgagag gaaagccagc tccccttctc ctctaactat tggaacgcca gaaagtcaga
     3061 gaaagccttc cattattctc agcaagtctc agctccagga tacattaata catctaataa
     3121 agaatgattc cagcttcctc agtacacttc atgaagtcta cttgcaggtg ctgaccaaga
     3181 acaaagacaa ccacaaccta tgagaattct gcagtcgacg gtaccgcggg cccgggatcc
     3241 accggatcta gataactgat cataatcagc cataccacat ttgtagaggt tttacttgct
     3301 ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt
     3361 gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     3421 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     3481 tcttaacgcg taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta
     3541 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga
     3601 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa
     3661 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga
     3721 accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc
     3781 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga
     3841 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg
     3901 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt
     3961 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     4021 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtcc
     4081 tgaggcggaa agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
     4141 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
     4201 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     4261 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     4321 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc
     4381 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaagat
     4441 cgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg
     4501 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg
     4561 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa
     4621 gaccgacctg tccggtgccc tgaatgaact gcaagacgag gcagcgcggc tatcgtggct
     4681 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga
     4741 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc
     4801 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac
     4861 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc
     4921 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
     4981 gttcgccagg ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga cccatggcga
     5041 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
     5101 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga
     5161 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
     5221 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg
     5281 ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc
     5341 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
     5401 cagcgcgggg atctcatgct ggagttcttc gcccacccta gggggaggct aactgaaaca
     5461 cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa
     5521 acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg ctggcactct
     5581 gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc cttttcccca
     5641 ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc
     5701 cctgccatag cctcaggtta ctcatatata ctttagattg atttaaaact tcatttttaa
     5761 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     5821 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     5881 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     5941 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     6001 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     6061 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     6121 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     6181 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     6241 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     6301 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     6361 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     6421 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     6481 tttttacggt tcctggcctt ttgctggcct tttgctca
//
