LOCUS       sequence-244485-        6963 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of Human TRIM28.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6963)
  AUTHORS   Li J, Xi Y, Li W, McCarthy RL, Stratton SA, Zou W, Li W, Dent SY, 
            Jain AK, Barton MC
  TITLE     TRIM28 interacts with EZH2 and SWI/SNF to activate genes that 
            promote mammosphere formation.
  JOURNAL   Oncogene. 2017 May 25;36(21):2991-3001. doi: 10.1038/onc.2016.453. 
            Epub 2017 Jan 9.
  PUBMED    28068325
REFERENCE   2  (bases 1 to 6963)
  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: 
            "10.1038/onc.2016.453"; journalName: "Oncogene"; date: "2017-05-25- 
            25"; volume: "36"; issue: "21"; pages: "2991-3001"
COMMENT     Sequence Label: Flag-TRIM28
FEATURES             Location/Qualifiers
     source          1..6963
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(42..64)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     164..183
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        328..707
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        708..911
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     862..882
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     956..975
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        956..974
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      988..997
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1000..1023
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1027..1050
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     intron          3708..3773
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             3903..3923
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    4195..4329
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4232..4251)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4286..4305
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      4787..4921
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4986..5003)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5157..5745)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5237..5256)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5916..6707,6708..6776))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6539..6558
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6777..6881)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     join(6949..6963,1..4)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc
       61 ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc
      121 gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc agagcagatt
      181 gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac
      241 cgcatcaggc gccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg
      301 gcctcttcgc tattacgcca gcgcgttgac attgattatt gactagttat taatagtaat
      361 caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg
      421 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt
      481 atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac
      541 ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg
      601 acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact
      661 ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt
      721 ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc
      781 ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc
      841 gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata
      901 taagcagagc tctctggcta actagagaac ccactgctta ctggcttatc gaaattaata
      961 cgactcacta tagggagacc caagcttgcc accatggcag actacaagga cgacgatgac
     1021 aaggcagact acaaggacga cgatgacaag ggtacctcta gaatggcggc ctccgcggcg
     1081 gcagcctcgg cagcagcggc ctcggccgcc tctggcagcc cgggcccggg cgagggctcc
     1141 gctggcggcg aaaagcgctc caccgcccct tcggccgcag cctcggcctc tgcctcagcc
     1201 gcggcgtcgt cgcccgcggg gggcggcgcc gaggcgctgg agctgctgga gcactgcggc
     1261 gtgtgcagag agcgcctgcg acccgagagg gagccccgcc tgctgccctg tttgcactcg
     1321 gcctgtagtg cctgcttagg gcccgcggcc cccgccgccg ccaacagctc gggggacggc
     1381 ggggcggcgg gcgacggcac cgtggtggac tgtcccgtgt gcaagcaaca gtgcttctcc
     1441 aaagacatcg tggagaatta tttcatgcgt gatagtggca gcaaggctgc caccgacgcc
     1501 caggatgcga accagtgctg cactagctgt gaggataatg ccccagccac cagctactgt
     1561 gtggagtgct cggagcctct gtgtgagacc tgtgtagagg cgcaccagcg ggtgaagtac
     1621 accaaggacc atactgtgcg ctctactggg ccagccaagt ctcgggatgg tgaacgtact
     1681 gtctattgca acgtacacaa gcatgaaccc cttgtgctgt tttgtgagag ctgtgatact
     1741 ctcacctgcc gagactgcca gctcaatgcc cacaaggacc accagtacca gttcttagag
     1801 gatgcagtga ggaaccagcg caagctcctg gcctcactgg tgaagcgcct tggggacaaa
     1861 catgcaacat tgcagaagag caccaaggag gttcgcagct caatccgcca ggtgtctgac
     1921 gtacagaagc gtgtgcaagt ggatgtcaag atggccatcc tgcagatcat gaaggagctg
     1981 aataagcggg gccgtgtgct ggtcaatgat gcccagaagg tgactgaggg gcagcaggag
     2041 cgcctggagc ggcagcactg gaccatgacc aagatccaga agcaccagga gcacattctg
     2101 cgctttgcct cttgggctct ggagagtgac aacaacacag cccttttgct ttctaagaag
     2161 ttgatctact tccagctgca ccgggccctc aagatgattg tggatcccgt ggagccacat
     2221 ggcgagatga agtttcagtg ggacctcaat gcctggacca agagtgccga ggcctttggc
     2281 aagattgtgg cagagcgtcc tggcactaac tcaacaggcc ctgcacccat ggcccctcca
     2341 agagccccag ggcccctgag caagcagggc tctggcagca gccagcccat ggaggtgcag
     2401 gaaggctatg gctttgggtc aggagatgat ccctactcaa gtgcagagcc ccatgtgtca
     2461 ggtgtgaaac ggtcccgctc aggtgagggc gaggtgagcg gccttatgcg caaggtgcca
     2521 cgagtgagcc ttgaacgcct ggacctggac ctcacagctg acagccagcc acccgtcttc
     2581 aaggtcttcc caggcagtac cactgaggac tacaacctta ttgttattga acgtggcgct
     2641 gccgctgcag ctaccggcca gccagggact gcgcctgcag gaacccctgg tgccccaccc
     2701 ctggctggca tggccattgt caaggaggag gagacggagg ctgccattgg agcccctcct
     2761 actgccactg agggccctga gaccaaacct gtgcttatgg ctcttgcgga gggtcctggt
     2821 gctgagggtc cccgcctggc ctcacctagt ggcagcacca gctcagggct ggaggtggtg
     2881 gctcctgagg gtacctcagc cccaggtggt ggcccgggaa ccctggatga cagtgccacc
     2941 atttgccgtg tctgccagaa gccaggcgat ctggttatgt gcaaccagtg tgagttttgt
     3001 ttccacctgg actgtcacct gccggccctg caggatgtac caggggagga gtggagctgc
     3061 tcactctgcc atgtgctccc tgacctgaag gaggaggatg gcagcctcag cctggatggt
     3121 gcagacagca ctggcgtggt ggccaagctc tcaccagcca accagcggaa atgtgagcgt
     3181 gtactgctgg ccctattctg tcacgaaccc tgccgccccc tgcatcagct ggctaccgac
     3241 tccaccttct ccctggacca gcccggtggc accctggatc tgaccctgat ccgtgcccgc
     3301 ctccaggaga agttgtcacc tccctacagc tccccacagg agtttgccca ggatgtgggc
     3361 cgcatgttca agcaattcaa caagttaact gaggacaagg cagacgtgca gtccatcatc
     3421 ggcctgcagc gcttcttcga gacgcgcatg aacgaggcct tcggtgacac caagttctct
     3481 gctgtgctgg tggagccccc gccgatgagc ctgcctggtg ctggcctgag ttcccaggag
     3541 ctgtctggtg gccctggtga tggcccctga ctcgagcggc cgcccgatat caccctgcag
     3601 ggatcctggc cagctagcta ggtagctaga ggatctttgt gaaggaacct tacttctgtg
     3661 gtgtgacata attggacaaa ctacctacag agatttaaag ctctaaggta aatataaaat
     3721 ttttaagtgt ataatgtgtt aaactactga ttctaattgt ttgtgtattt tagattccaa
     3781 cctatggaac tgatgaatgg gagcagtggt ggaatgcctt taatgaggaa aacctgtttt
     3841 gctcagaaga aatgccatct agtgatgatg aggctactgc tgactctcaa cattctactc
     3901 ctccaaaaaa gaagagaaag gtagaagacc ccaaggactt tccttcagaa ttgctaagtt
     3961 ttttgagtca tgctgtgttt agtaatagaa ctcttgcttg ctttgctatt tacaccacaa
     4021 aggaaaaagc tgcactgcta tacaagaaaa ttatggaaaa atatttgatg tatagtgcct
     4081 tgactagaga tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac
     4141 ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg
     4201 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     4261 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     4321 gtctggatca gcttcagaag atggcggagg gcctccaaca cagtaatttt cctcccgact
     4381 cttaaaatag aaaatgtcaa gtcagttaag caggaagtga ctaactgacc gcagctggcc
     4441 gtgcgacatc ctcttttaat tagttgctag gcaactgccc tccagagggc agtgtggttt
     4501 tgcaagagga agcaaaaagc ctctccaccc aggcctagaa tgtttccacc caatcattac
     4561 tatgacaaca gctgtttttt ttagtattaa gcagaggccg ggggcccctg gcctccgctt
     4621 actctggaga aaaagaagag aggcattgta gaggcttcca gaggcaactt gtcaaaacag
     4681 gactgcttct atttctgtca cactgtctgg ccctgtcaca aggtccagca cctccatacc
     4741 ccctttaata agcagtttgg gaacgggtgc gggtcttact ccgcccatcc cgcccctaac
     4801 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     4861 ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     4921 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     4981 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     5041 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     5101 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     5161 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     5221 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     5281 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     5341 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     5401 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     5461 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     5521 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     5581 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     5641 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     5701 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     5761 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     5821 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     5881 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     5941 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     6001 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     6061 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     6121 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     6181 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     6241 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     6301 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     6361 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     6421 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     6481 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     6541 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     6601 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     6661 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     6721 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     6781 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     6841 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     6901 ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc
     6961 acg
//
