LOCUS       sequence-11550-e        6296 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 6296)
  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 6296)
  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-HsEDC3
FEATURES             Location/Qualifiers
     source          1..6296
                     /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    3032..3153
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3069..3088)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3123..3142
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3160..3615)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3297..3318)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3509..3528
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3642..3746
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3748..4105
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3748..3939
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3771..3791)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3956..4091
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4140..4934
                     /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             4140..4934
                     /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(4194..4213)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4804..4823
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5122..5141)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5166..5213
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5542..6130
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6031..6050
                     /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 atccatggct acagattggc tgggaagtat tgtgtccatc
     1381 aattgtggag atagcttggg tgtctatcag ggaagagtgt cagctgtgga tcaggtcagc
     1441 cagaccattt ctctcacccg gcctttccat aatggagtga agtgtcttgt tccagaagtc
     1501 accttcaggg caggtgacat tacggagtta aaaattctgg agataccagg acctggagac
     1561 aaccaacatt ttggagacct tcatcaaaca gaattaggcc cctctggtgc tggctgccaa
     1621 gtgggcatca atcagaatgg cacaggcaag tttgtcaaga agccagcctc ttccagcagt
     1681 gcccctcaga atatccctaa gaggacagat gtgaagagcc aggatgttgc cgtttccccg
     1741 cagcagcaac agtgctcaaa gagctatgtc gacaggcaca tggaatcctt gagtcagtcc
     1801 aaaagtttcc gtcgtcggca caactcctgg tcatctagta gcaggcaccc aaatcaggca
     1861 actcccaaga aaagtggttt aaagaatggc cagatgaaga ataaagatga cgagtgcttc
     1921 ggggatgata ttgaggagat cccagacaca gattttgatt ttgaagggaa cctggctctt
     1981 tttgacaagg cagctgtgtt tgaggagatt gatacctatg aaaggagaag tggtacccgt
     2041 tcccggggca tcccaaatga aaggcccact cggtaccgcc atgatgagaa catcttggag
     2101 tccgagccca ttgtctatcg acggatcata gtgccccaca acgtgagcaa ggagttctgc
     2161 acggactctg gcctggttgt cccaagtatt tcctatgagc tgcataaaaa gctgttgtcc
     2221 gtggctgaga agcatgggct gacccttgag cggagactgg agatgacagg tgtgtgtgcc
     2281 agtcagatgg cactgaccct cctcggagga cctaacaggt tgaatcccaa aaatgttcac
     2341 cagaggccta cagtggctct actgtgtgga cctcatgtga agggggctca gggtatcagc
     2401 tgtggaaggc acctagccaa ccatgatgtc caggtcatcc ttttcctgcc caattttgtc
     2461 aagatgttgg aatctatcac caatgagctg tcgctcttca gcaagaccca aggccaacaa
     2521 gtgtctagcc tcaaagatct gcccactagc cctgtggacc tggtcatcaa ctgcctggat
     2581 tgccctgaga acgtcttcct gcgcgatcaa ccctggtaca aggcagctgt ggcctgggcc
     2641 aaccagaacc gggcaccagt actcagcata gaccctcctg tgcatgaagt cgaacagggc
     2701 attgatgcca aatggtcact ggcactgggc ctgcctctgc cactggggga gcacgcaggc
     2761 cgtatctatt tgtgcgacat tggcattccc cagcaggtct tccaggaggt gggcatcaac
     2821 taccactcgc cctttggctg caagtttgtt atcccactgc actctgctta ggaattctgc
     2881 agtcgacggt accgcgggcc cgggatccac cggatctaga taactgatca taatcagcca
     2941 taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct
     3001 gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta
     3061 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     3121 ttgtggtttg tccaaactca tcaatgtatc ttaacgcgta aattgtaagc gttaatattt
     3181 tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa
     3241 tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag
     3301 tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg
     3361 tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga
     3421 ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg
     3481 gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg
     3541 cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc
     3601 cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     3661 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     3721 ttcaataata ttgaaaaagg aagagtcctg aggcggaaag aaccagctgt ggaatgtgtg
     3781 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     3841 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     3901 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
     3961 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     4021 ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt
     4081 ttttggaggc ctaggctttt gcaaagatcg atcaagagac aggatgagga tcgtttcgca
     4141 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg
     4201 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag
     4261 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc
     4321 aagacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc
     4381 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg
     4441 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc
     4501 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca
     4561 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag
     4621 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgagc atgcccgacg
     4681 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg
     4741 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca
     4801 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc
     4861 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg
     4921 acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct
     4981 gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
     5041 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     5101 ccaccctagg gggaggctaa ctgaaacacg gaaggagaca ataccggaag gaacccgcgc
     5161 tatgacggca ataaaaagac agaataaaac gcacggtgtt gggtcgtttg ttcataaacg
     5221 cggggttcgg tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca
     5281 atacgcccgc gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg
     5341 gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc tcaggttact catatatact
     5401 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     5461 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     5521 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     5581 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     5641 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta
     5701 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     5761 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     5821 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     5881 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     5941 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     6001 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     6061 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     6121 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     6181 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgccat
     6241 gcattagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatg
//
