LOCUS       sequence-251933-        6175 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of Human TRIM28 with deleted PHD and BROMO 
            domain.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6175)
  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 6175)
  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: HA-TRIM28deltaPB
FEATURES             Location/Qualifiers
     source          1..6175
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..978
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     promoter        1013..1031
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1013..1030
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             1066..1146
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAYPYDVPDYAYPYDVPDYA"
     polyA_signal    3205..3339
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3229..3248)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3283..3302
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3452..3472)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3452..3470)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3491..3507)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3491..3507)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3504..3526)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3515..3531
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(3539..3545,3546..3563,3564..3569))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3584..3605
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3722..3739)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3893..4481)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3973..3992)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4652..5443,5444..5512))
                     /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     5275..5294
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5513..5617)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5643..6098)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5780..5801)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5992..6011
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gccaattcaa tatggcgtat atggactcat gccaattcaa tatggtggat ctggacctgt
       61 gccaattcaa tatggcgtat atggactcgt gccaattcaa tatggtggat ctggacccca
      121 gccaattcaa tatggcggac ttggcaccat gccaattcaa tatggcggac ctggcactgt
      181 gccaactggg gaggggtcta cttggcacgg tgccaagttt gaggaggggt cttggccctg
      241 tgccaagtcc gccatattga attggcatgg tgccaataat ggcggccata ttggctatat
      301 gccaggatca atatataggc aatatccaat atggccctat gccaatatgg ctattggcca
      361 ggttcaatac tatgtattgg ccctatgcca tatagtattc catatatggg ttttcctatt
      421 gacgtagata gcccctccca atgggcggtc ccatatacca tatatggggc ttcctaatac
      481 cgcccatagc cactccccca ttgacgtcaa tggtctctat atatggtctt tcctattgac
      541 gtcatatggg cggtcctatt gacgtatatg gcgcctcccc cattgacgtc aattacggta
      601 aatggcccgc ctggctcaat gcccattgac gtcaatagga ccacccacca ttgacgtcaa
      661 tgggatggct cattgcccat tcatatccgt tctcacgccc cctattgacg tcaatgacgg
      721 taaatggccc acttggcagt acatcaatat ctattaatag taacttggca agtacattac
      781 tattggaagt acgccagggt acattggcag tactcccatt gacgtcaatg gcggtaaatg
      841 gcccgcgatg gctgccaagt acatccccat tgacgtcaat ggggaggggc aatgacgcaa
      901 atgggcgttc cattgacgta aatgggcggt aggcgtgcct aatgggaggt ctatataagc
      961 aatgctcgtt tagggaaccg ccattctgcc tggggacgtc ggagcaagct tgatttaggt
     1021 gacactatag aatacaagct acttgttctt tttgcaggat ccatgtaccc atacgatgtt
     1081 ccagattacg cttatcctta tgacgtacct gactatgcat acccttatga tgtaccagac
     1141 tacgctggcc ggccaatggc ggcctccgcg gcggcagcct cggcagcagc ggcctcggcc
     1201 gcctctggca gcccgggccc gggcgagggc tccgctggcg gcgaaaagcg ctccaccgcc
     1261 ccttcggccg cagcctcggc ctctgcctca gccgcggcgt cgtcgcccgc ggggggcggc
     1321 gccgaggcgc tggagctgct ggagcactgc ggcgtgtgca gagagcgcct gcgacccgag
     1381 agggagcccc gcctgctgcc ctgtttgcac tcggcctgta gtgcctgctt agggcccgcg
     1441 gcccccgccg ccgccaacag ctcgggggac ggcggggcgg cgggcgacgg caccgtggtg
     1501 gactgtcccg tgtgcaagca acagtgcttc tccaaagaca tcgtggagaa ttatttcatg
     1561 cgtgatagtg gcagcaaggc tgccaccgac gcccaggatg cgaaccagtg ctgcactagc
     1621 tgtgaggata atgccccagc caccagctac tgtgtggagt gctcggagcc tctgtgtgag
     1681 acctgtgtag aggcgcacca gcgggtgaag tacaccaagg accatactgt gcgctctact
     1741 gggccagcca agtctcggga tggtgaacgt actgtctatt gcaacgtaca caagcatgaa
     1801 ccccttgtgc tgttttgtga gagctgtgat actctcacct gccgagactg ccagctcaat
     1861 gcccacaagg accaccagta ccagttctta gaggatgcag tgaggaacca gcgcaagctc
     1921 ctggcctcac tggtgaagcg ccttggggac aaacatgcaa cattgcagaa gagcaccaag
     1981 gaggttcgca gctcaatccg ccaggtgtct gacgtacaga agcgtgtgca agtggatgtc
     2041 aagatggcca tcctgcagat catgaaggag ctgaataagc ggggccgtgt gctggtcaat
     2101 gatgcccaga aggtgactga ggggcagcag gagcgcctgg agcggcagca ctggaccatg
     2161 accaagatcc agaagcacca ggagcacatt ctgcgctttg cctcttgggc tctggagagt
     2221 gacaacaaca cagccctttt gctttctaag aagttgatct acttccagct gcaccgggcc
     2281 ctcaagatga ttgtggatcc cgtggagcca catggcgaga tgaagtttca gtgggacctc
     2341 aatgcctgga ccaagagtgc cgaggccttt ggcaagattg tggcagagcg tcctggcact
     2401 aactcaacag gccctgcacc catggcccct ccaagagccc cagggcccct gagcaagcag
     2461 ggctctggca gcagccagcc catggaggtg caggaaggct atggctttgg gtcaggagat
     2521 gatccctact caagtgcaga gccccatgtg tcaggtgtga aacggtcccg ctcaggtgag
     2581 ggcgaggtga gcggccttat gcgcaaggtg ccacgagtga gccttgaacg cctggacctg
     2641 gacctcacag ctgacagcca gccacccgtc ttcaaggtct tcccaggcag taccactgag
     2701 gactacaacc ttattgttat tgaacgtggc gctgccgctg cagctaccgg ccagccaggg
     2761 actgcgcctg caggaacccc tggtgcccca cccctggctg gcatggccat tgtcaaggag
     2821 gaggagacgg aggctgccat tggagcccct cctactgcca ctgagggccc tgagaccaaa
     2881 cctgtgctta tggctcttgc ggagggtcct ggtgctgagg gtccccgcct ggcctcacct
     2941 agtggcagca ccagctcagg gctggaggtg gtggctcctg agggtacctc agccccaggt
     3001 ggtggcccgg gaaccctgga tgacagtgcc accattacca agttctcgat gacagtgcca
     3061 ccattaccaa gttctctgct gtgctggtgg agcccccgcc gatgagcctg cctggtgctg
     3121 gcctgagttc ccaggagctg tctggtggcc ctggtgatgg cccctgaggc gcgcctctag
     3181 aactatagtg agtcgtatta cgtagatcca gacatgataa gatacattga tgagtttgga
     3241 caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt
     3301 gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat
     3361 tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt aattcgcggc cgcggcgcca
     3421 atgcattggg cccggtaccc agcttttgtt ccctttagtg agggttaatt gcgcgcttgg
     3481 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     3541 acatacgagc cgggagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     3601 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     3661 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     3721 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     3781 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     3841 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     3901 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     3961 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     4021 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     4081 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     4141 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     4201 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     4261 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     4321 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     4381 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     4441 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     4501 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     4561 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     4621 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     4681 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     4741 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     4801 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     4861 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     4921 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     4981 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     5041 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     5101 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     5161 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     5221 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     5281 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     5341 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     5401 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     5461 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     5521 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     5581 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     5641 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
     5701 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
     5761 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
     5821 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
     5881 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
     5941 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
     6001 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
     6061 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
     6121 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagt cgacc
//
