LOCUS       sequence-235547-        7225 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 7225)
  AUTHORS   Liang Q, Deng H, Li X, Wu X, Tang Q, Chang TH, Peng H, Rauscher FJ 
            3rd, Ozato K, Zhu F
  TITLE     Tripartite motif-containing protein 28 is a small ubiquitin-related 
            modifier E3 ligase and negative regulator of IFN regulatory factor 
            7.
  JOURNAL   J Immunol. 2011 Nov 1;187(9):4754-63. doi: 10.4049/jimmunol.1101704.
            Epub 2011 Sep 21.
  PUBMED    21940674
REFERENCE   2  (bases 1 to 7225)
  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: 
            "10.4049/jimmunol.1101704"; journalName: "J Immunol"; date: 
            "2011-11-1- 1"; volume: "187"; issue: "9"; pages: "4754-63"
COMMENT     Sequence Label: pEGFP-TRIM28
FEATURES             Location/Qualifiers
     source          1..7225
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      547..556
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(553..555,556..558,559..1269)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGESEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(598..619)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(859..878)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1206..1227
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     regulatory      1283..1292
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    3953..4074
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3990..4009)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4044..4063
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4081..4536)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4218..4239)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4430..4449
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4563..4667
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4669..5026
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4669..4860
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4692..4712)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4877..5012
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5061..5855
                     /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             5061..5855
                     /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(5115..5134)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5725..5744
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6043..6062)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6087..6134
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6463..7051
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6952..6971
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      541 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
      601 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gagcgagggc
      661 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
      721 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
      781 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
      841 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
      901 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
      961 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     1021 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     1081 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1141 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
     1201 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     1261 ctgtacaagt ccggactcag atccatggcg gcctccgcgg cggcagcctc ggcagcagcg
     1321 gcctcggccg cctctggcag cccgggcccg ggcgagggct ccgctggcgg cgaaaagcgc
     1381 tccaccgccc cttcggccgc agcctcggcc tctgcctcag ccgcggcgtc gtcgcccgcg
     1441 gggggcggcg ccgaggcgct ggagctgctg gagcactgcg gcgtgtgcag agagcgcctg
     1501 cgacccgaga gggagccccg cctgctgccc tgtttgcact cggcctgtag tgcctgctta
     1561 gggcccgcgg cccccgccgc cgccaacagc tcgggggacg gcggggcggc gggcgacggc
     1621 accgtggtgg actgtcccgt gtgcaagcaa cagtgcttct ccaaagacat cgtggagaat
     1681 tatttcatgc gtgatagtgg cagcaaggct gccaccgacg cccaggatgc gaaccagtgc
     1741 tgcactagct gtgaggataa tgccccagcc accagctact gtgtggagtg ctcggagcct
     1801 ctgtgtgaga cctgtgtaga ggcgcaccag cgggtgaagt acaccaagga ccatactgtg
     1861 cgctctactg ggccagccaa gtctcgggat ggtgaacgta ctgtctattg caacgtacac
     1921 aagcatgaac cccttgtgct gttttgtgag agctgtgata ctctcacctg ccgagactgc
     1981 cagctcaatg cccacaagga ccaccagtac cagttcttag aggatgcagt gaggaaccag
     2041 cgcaagctcc tggcctcact ggtgaagcgc cttggggaca aacatgcaac attgcagaag
     2101 agcaccaagg aggttcgcag ctcaatccgc caggtgtctg acgtacagaa gcgtgtgcaa
     2161 gtggatgtca agatggccat cctgcagatc atgaaggagc tgaataagcg gggccgtgtg
     2221 ctggtcaatg atgcccagaa ggtgactgag gggcagcagg agcgcctgga gcggcagcac
     2281 tggaccatga ccaagatcca gaagcaccag gagcacattc tgcgctttgc ctcttgggct
     2341 ctggagagtg acaacaacac agcccttttg ctttctaaga agttgatcta cttccagctg
     2401 caccgggccc tcaagatgat tgtggatccc gtggagccac atggcgagat gaaatttcag
     2461 tgggacctca atgcctggac caagagtgcc gaggcctttg gcaagattgt ggcagagcgt
     2521 cctggcacta actcaacagg ccctgcaccc atggcccctc caagagcccc agggcccctg
     2581 agcaagcagg gctctggcag cagccagccc atggaggtgc aggaaggcta tggctttggg
     2641 tcaggagatg atccctactc aagtgcagag ccccatgtgt caggtgtgaa acggtcccgc
     2701 tcaggtgagg gcgaggtgag cggccttatg cgcaaggtgc cacgagtgag ccttgaacgc
     2761 ctggacctgg acctcacagc tgacagccag ccacccgtct tcaaggtctt cccaggcagt
     2821 accactgagg actacaacct tattgttatt gaacgtggcg ctgccgctgc agctaccggc
     2881 cagccaggga ctgcgcctgc aggaacccct ggtgccccac ccctggctgg catggccatt
     2941 gtcaaggagg aggagacgga ggctgccatt ggagcccctc ctactgccac tgagggccct
     3001 gagaccaaac ctgtgcttat ggctcttgcg gagggtcctg gtgctgaggg tccccgcctg
     3061 gcctcaccta gtggcagcac cagctcaggg ctggaggtgg tggctcctga gggtacctca
     3121 gccccaggtg gtggcccggg aaccctggat gacagtgcca ccatttgccg tgtctgccag
     3181 aagccaggcg atctggttat gtgcaaccag tgtgagtttt gtttccacct ggactgtcac
     3241 ctgccggccc tgcaggatgt accaggggag gagtggagct gctcactctg ccatgtgctc
     3301 cctgacctga aggaggagga tggcagcctc agcctggatg gtgcagacag cactggcgtg
     3361 gtggccaagc tctcaccagc caaccagcgg aaatgtgagc gtgtactgct ggccctattc
     3421 tgtcacgaac cctgccgccc cctgcatcag ctggctaccg actccacctt ctccctggac
     3481 cagcccggtg gcaccctgga tctgaccctg atccgtgccc gcctccagga gaagttgtca
     3541 cctccctaca gctccccaca ggagtttgcc caggatgtgg gccgcatgtt caagcaattc
     3601 aacaagttaa ctgaggacaa ggcagacgtg cagtccatca tcggcctgca gcgcttcttc
     3661 gagacgcgca tgaacgaggc cttcggtgac accaagttct ctgctgtgct ggtggagccc
     3721 ccgccgatga gcctgcctgg tgctggcctg agttcccagg agctgtctgg tggccctggt
     3781 gatggcccct gagaattctg cagtcgacgg taccgcgggc ccgggatcca ccggatctag
     3841 ataactgatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct
     3901 cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt
     3961 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     4021 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttaacgcgt
     4081 aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat
     4141 tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga
     4201 tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca
     4261 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct
     4321 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc
     4381 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag
     4441 cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca
     4501 cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt cggggaaatg
     4561 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     4621 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtcct gaggcggaaa
     4681 gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
     4741 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
     4801 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
     4861 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     4921 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt
     4981 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaagatc gatcaagaga
     5041 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg
     5101 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
     5161 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
     5221 ccggtgccct gaatgaactg caagacgagg cagcgcggct atcgtggctg gccacgacgg
     5281 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
     5341 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
     5401 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
     5461 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
     5521 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
     5581 tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     5641 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     5701 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
     5761 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     5821 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac
     5881 cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga
     5941 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     6001 tctcatgctg gagttcttcg cccaccctag ggggaggcta actgaaacac ggaaggagac
     6061 aataccggaa ggaacccgcg ctatgacggc aataaaaaga cagaataaaa cgcacggtgt
     6121 tgggtcgttt gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc
     6181 accgagaccc cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc
     6241 aagttcgggt gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc
     6301 ctcaggttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     6361 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     6421 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     6481 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     6541 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     6601 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     6661 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     6721 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     6781 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     6841 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     6901 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     6961 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     7021 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     7081 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     7141 ggataaccgt attaccgcca tgcattagtt attaatagta atcaattacg gggtcattag
     7201 ttcatagccc atatatggag ttccg
//
