LOCUS       sequence-243631-        6479 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of Human TRIM28 with deleted RING domain.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6479)
  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 6479)
  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-TRIM28deltaRING
FEATURES             Location/Qualifiers
     source          1..6479
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        37..1014
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     promoter        1049..1067
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1049..1066
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             1102..1182
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAYPYDVPDYAYPYDVPDYA"
     polyA_signal    3545..3679
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3569..3588)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3623..3642
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3792..3812)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3792..3810)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3831..3847)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3831..3847)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3844..3866)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3855..3871
                     /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(3879..3885,3886..3903,3904..3909))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3924..3945
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4062..4079)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4233..4821)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4313..4332)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4992..5783,5784..5852))
                     /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     5615..5634
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5853..5957)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5983..6438)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6120..6141)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6332..6351
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 tcggtgcggg cctcttcgct attacgccag tcgaccgcca attcaatatg gcgtatatgg
       61 actcatgcca attcaatatg gtggatctgg acctgtgcca attcaatatg gcgtatatgg
      121 actcgtgcca attcaatatg gtggatctgg accccagcca attcaatatg gcggacttgg
      181 caccatgcca attcaatatg gcggacctgg cactgtgcca actggggagg ggtctacttg
      241 gcacggtgcc aagtttgagg aggggtcttg gccctgtgcc aagtccgcca tattgaattg
      301 gcatggtgcc aataatggcg gccatattgg ctatatgcca ggatcaatat ataggcaata
      361 tccaatatgg ccctatgcca atatggctat tggccaggtt caatactatg tattggccct
      421 atgccatata gtattccata tatgggtttt cctattgacg tagatagccc ctcccaatgg
      481 gcggtcccat ataccatata tggggcttcc taataccgcc catagccact cccccattga
      541 cgtcaatggt ctctatatat ggtctttcct attgacgtca tatgggcggt cctattgacg
      601 tatatggcgc ctcccccatt gacgtcaatt acggtaaatg gcccgcctgg ctcaatgccc
      661 attgacgtca ataggaccac ccaccattga cgtcaatggg atggctcatt gcccattcat
      721 atccgttctc acgcccccta ttgacgtcaa tgacggtaaa tggcccactt ggcagtacat
      781 caatatctat taatagtaac ttggcaagta cattactatt ggaagtacgc cagggtacat
      841 tggcagtact cccattgacg tcaatggcgg taaatggccc gcgatggctg ccaagtacat
      901 ccccattgac gtcaatgggg aggggcaatg acgcaaatgg gcgttccatt gacgtaaatg
      961 ggcggtaggc gtgcctaatg ggaggtctat ataagcaatg ctcgtttagg gaaccgccat
     1021 tctgcctggg gacgtcggag caagcttgat ttaggtgaca ctatagaata caagctactt
     1081 gttctttttg caggatccat gtacccatac gatgttccag attacgctta tccttatgac
     1141 gtacctgact atgcataccc ttatgatgta ccagactacg ctggccggcc aatggcggcc
     1201 tccgcggcgg cagcctcggc agcagcggcc tcggccgcct ctggcagccc gggcccgggc
     1261 gagggctccg ctggcggcga aaagcgctcc accgcccctt cggccgcagc ctcggcctct
     1321 gcctcagccg cggcgtcgtc gcccgcgggg ggcggcgccg aggcgctgga gttctccaaa
     1381 gacatcgtgg agaattattt catgcgtgat agtggcagca aggctgccac cgacgcccag
     1441 gatgcgaacc agtgctgcac tagctgtgag gataatgccc cagccaccag ctactgtgtg
     1501 gagtgctcgg agcctctgtg tgagacctgt gtagaggcgc accagcgggt gaagtacacc
     1561 aaggaccata ctgtgcgctc tactgggcca gccaagtctc gggatggtga acgtactgtc
     1621 tattgcaacg tacacaagca tgaacccctt gtgctgtttt gtgagagctg tgatactctc
     1681 acctgccgag actgccagct caatgcccac aaggaccacc agtaccagtt cttagaggat
     1741 gcagtgagga accagcgcaa gctcctggcc tcactggtga agcgccttgg ggacaaacat
     1801 gcaacattgc agaagagcac caaggaggtt cgcagctcaa tccgccaggt gtctgacgta
     1861 cagaagcgtg tgcaagtgga tgtcaagatg gccatcctgc agatcatgaa ggagctgaat
     1921 aagcggggcc gtgtgctggt caatgatgcc cagaaggtga ctgaggggca gcaggagcgc
     1981 ctggagcggc agcactggac catgaccaag atccagaagc accaggagca cattctgcgc
     2041 tttgcctctt gggctctgga gagtgacaac aacacagccc ttttgctttc taagaagttg
     2101 atctacttcc agctgcaccg ggccctcaag atgattgtgg atcccgtgga gccacatggc
     2161 gagatgaagt ttcagtggga cctcaatgcc tggaccaaga gtgccgaggc ctttggcaag
     2221 attgtggcag agcgtcctgg cactaactca acaggccctg cacccatggc ccctccaaga
     2281 gccccagggc ccctgagcaa gcagggctct ggcagcagcc agcccatgga ggtgcaggaa
     2341 ggctatggct ttgggtcagg agatgatccc tactcaagtg cagagcccca tgtgtcaggt
     2401 gtgaaacggt cccgctcagg tgagggcgag gtgagcggcc ttatgcgcaa ggtgccacga
     2461 gtgagccttg aacgcctgga cctggacctc acagctgaca gccagccacc cgtcttcaag
     2521 gtcttcccag gcagtaccac tgaggactac aaccttattg ttattgaacg tggcgctgcc
     2581 gctgcagcta ccggccagcc agggactgcg cctgcaggaa cccctggtgc cccacccctg
     2641 gctggcatgg ccattgtcaa ggaggaggag acggaggctg ccattggagc ccctcctact
     2701 gccactgagg gccctgagac caaacctgtg cttatggctc ttgcggaggg tcctggtgct
     2761 gagggtcccc gcctggcctc acctagtggc agcaccagct cagggctgga ggtggtggct
     2821 cctgagggta cctcagcccc aggtggtggc ccgggaaccc tggatgacag tgccaccatt
     2881 tgccgtgtct gccagaagcc aggcgatctg gttatgtgca accagtgtga gttttgtttc
     2941 cacctggact gtcacctgcc ggccctgcag gatgtaccag gggaggagtg gagctgctca
     3001 ctctgccatg tgctccctga cctgaaggag gaggatggca gcctcagcct ggatggtgca
     3061 gacagcactg gcgtggtggc caagctctca ccagccaacc agcggaaatg tgagcgtgta
     3121 ctgctggccc tattctgtca cgaaccctgc cgccccctgc atcagctggc taccgactcc
     3181 accttctccc tggaccagcc cggtggcacc ctggatctga ccctgatccg tgcccgcctc
     3241 caggagaagt tgtcacctcc ctacagctcc ccacaggagt ttgcccagga tgtgggccgc
     3301 atgttcaagc aattcaacaa gttaactgag gacaaggcag acgtgcagtc catcatcggc
     3361 ctgcagcgct tcttcgagac gcgcatgaac gaggccttcg gtgacaccaa gttctctgct
     3421 gtgctggtgg agcccccgcc gatgagcctg cctggtgctg gcctgagttc ccaggagctg
     3481 tctggtggcc ctggtgatgg cccctgaggc gcgcctctag aactatagtg agtcgtatta
     3541 cgtagatcca gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca
     3601 gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat
     3661 aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg
     3721 ggaggtgtgg gaggtttttt aattcgcggc cgcggcgcca atgcattggg cccggtaccc
     3781 agcttttgtt ccctttagtg agggttaatt gcgcgcttgg cgtaatcatg gtcatagctg
     3841 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cgggagcata
     3901 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     3961 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     4021 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     4081 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     4141 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     4201 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     4261 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     4321 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     4381 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     4441 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     4501 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     4561 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     4621 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     4681 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     4741 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     4801 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     4861 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     4921 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     4981 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     5041 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     5101 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     5161 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     5221 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     5281 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     5341 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     5401 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     5461 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     5521 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     5581 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     5641 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     5701 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     5761 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     5821 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     5881 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     5941 caaatagggg ttccgcgcac atttccccga aaagtgccac ctaaattgta agcgttaata
     6001 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg
     6061 aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc
     6121 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa
     6181 ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt
     6241 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac
     6301 ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta
     6361 gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg
     6421 cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg caactgttgg gaagggcga
//
