LOCUS       sequence-243625-        5951 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of Human TRIM28 with deleted Bbox and CC 
            domains.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5951)
  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 5951)
  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-TRIM28deltaBCC
FEATURES             Location/Qualifiers
     source          1..5951
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        40..1024
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     promoter        1059..1077
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1059..1076
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             1112..1192
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAYPYDVPDYAYPYDVPDYA"
     polyA_signal    3027..3161
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3051..3070)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3105..3124
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3274..3294)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3274..3292)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3313..3329)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3313..3329)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3326..3348)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3337..3353
                     /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(3361..3367,3368..3385,3386..3391))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3406..3427
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3544..3561)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3715..4303)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3795..3814)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4474..5265,5266..5334))
                     /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     5097..5116
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5335..5439)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5465..5920)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5602..5623)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5814..5833
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 ggaagggcga tcggtgcggg cctcttcgct attacgccag tcgaccgcca attcaatatg
       61 gcgtatatgg actcatgcca attcaatatg gtggatctgg acctgtgcca attcaatatg
      121 gcgtatatgg actcgtgcca attcaatatg gtggatctgg accccagcca attcaatatg
      181 gcggacttgg caccatgcca attcaatatg gcggacctgg cactgtgcca actggggagg
      241 ggtctacttg gcacggtgcc aagtttgagg aggggtcttg gccctgtgcc aagtccgcca
      301 tattgaattg gcatggtgcc aataatggcg gccatattgg ctatatgcca ggatcaatat
      361 ataggcaata tccaatatgg ccctatgcca atatggctat tggccaggtt caatactatg
      421 tattggccct atgccatata gtattccata tatgggtttt cctattgacg tagatagccc
      481 ctcccaatgg gcggtcccat ataccatata tggggcttcc taataccgcc catagccact
      541 cccccattga cgtcaatggt ctctatatat ggtctttcct attgacgtca tatgggcggt
      601 cctattgacg tatatggcgc ctcccccatt gacgtcaatt acggtaaatg gcccgcctgg
      661 ctcaatgccc attgacgtca ataggaccac ccaccattga cgtcaatggg atggctcatt
      721 gcccattcat atccgttctc acgcccccta ttgacgtcaa tgacggtaaa tggcccactt
      781 ggcagtacat caatatctat taatagtaac ttggcaagta cattactatt ggaagtacgc
      841 cagggtacat tggcagtact cccattgacg tcaatggcgg taaatggccc gcgatggctg
      901 ccaagtacat ccccattgac gtcaatgggg aggggcaatg acgcaaatgg gcgttccatt
      961 gacgtaaatg ggcggtaggc gtgcctaatg ggaggtctat ataagcaatg ctcgtttagg
     1021 gaaccgccat tctgcctggg gacgtcggag caagcttgat ttaggtgaca ctatagaata
     1081 caagctactt gttctttttg caggatccat gtacccatac gatgttccag attacgctta
     1141 tccttatgac gtacctgact atgcataccc ttatgatgta ccagactacg ctggccggcc
     1201 aatggcggcc tccgcggcgg cagcctcggc agcagcggcc tcggccgcct ctggcagccc
     1261 gggcccgggc gagggctccg ctggcggcga aaagcgctcc accgcccctt cggccgcagc
     1321 ctcggcctct gcctcagccg cggcgtcgtc gcccgcgggg ggcggcgccg aggcgctgga
     1381 gctgctggag cactgcggcg tgtgcagaga gcgcctgcga cccgagaggg agccccgcct
     1441 gctgccctgt ttgcactcgg cctgtagtgc ctgcttaggg cccgcggccc ccgccgccgc
     1501 caacagctcg ggggacggcg gggcggcggg cgacggcacc gtggtggact gtcccgtgtg
     1561 caagcaacag tgcttctcca aagacatcgt ggagaattat ttcatgcgtg atagtattgt
     1621 ggatcccgtg gagccacatg gcgagatgaa gtttcagtgg gacctcaatg cctggaccaa
     1681 gagtgccgag gcctttggca agattgtggc agagcgtcct ggcactaact caacaggccc
     1741 tgcacccatg gcccctccaa gagccccagg gcccctgagc aagcagggct ctggcagcag
     1801 ccagcccatg gaggtgcagg aaggctatgg ctttgggtca ggagatgatc cctactcaag
     1861 tgcagagccc catgtgtcag gtgtgaaacg gtcccgctca ggtgagggcg aggtgagcgg
     1921 ccttatgcgc aaggtgccac gagtgagcct tgaacgcctg gacctggacc tcacagctga
     1981 cagccagcca cccgtcttca aggtcttccc aggcagtacc actgaggact acaaccttat
     2041 tgttattgaa cgtggcgctg ccgctgcagc taccggccag ccagggactg cgcctgcagg
     2101 aacccctggt gccccacccc tggctggcat ggccattgtc aaggaggagg agacggaggc
     2161 tgccattgga gcccctccta ctgccactga gggccctgag accaaacctg tgcttatggc
     2221 tcttgcggag ggtcctggtg ctgagggttc ccgcctggcc tcacctagtg gcagcaccag
     2281 ctcagggctg gaggtggtgg ctcctgaggg tacctcagcc ccaggtggtg gcccgggaac
     2341 cctggatgac agtgccacca tttgccgtgt ctgccagaag ccaggcgatc tggttatgtg
     2401 caaccagtgt gagttttgtt tccacctgga ctgtcacctg ccggccctgc aggatgtacc
     2461 aggggaggag tggagctgct cactctgcca tgtgctccct gacctgaagg aggaggatgg
     2521 cagcctcagc ctggatggtg cagacagcac tggcgtggtg gccaagctct caccagccaa
     2581 ccagcggaaa tgtgagcgtg tactgctggc cctattctgt cacgaaccct gccgccccct
     2641 gcatcagctg gctaccgact ccaccttctc cctggaccag cccggtggca ccctggatct
     2701 gaccctgatc cgtgcccgcc tccaggagaa gttgtcacct ccctacagct ccccacagga
     2761 gtttgcccag gatgtgggcc gcatgttcaa gcaattcaac aagttaactg aggacaaggc
     2821 agacgtgcag tccatcatcg gcctgcagcg cttcttcgag acgcgcatga acgaggcctt
     2881 cggtgacacc aagttctctg ctgtgctggt ggagcccccg ccgatgagcc tgcctggtgc
     2941 tggcctgagt tcccaggagc tgtctggtgg ccctggtgat ggcccctgag gcgcgcctct
     3001 agaactatag tgagtcgtat tacgtagatc cagacatgat aagatacatt gatgagtttg
     3061 gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta
     3121 ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc
     3181 attttatgtt tcaggttcag ggggaggtgt gggaggtttt ttaattcgcg gccgcggcgc
     3241 caatgcattg ggcccggtac ccagcttttg ttccctttag tgagggttaa ttgcgcgctt
     3301 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     3361 caacatacga gccgggagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     3421 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     3481 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     3541 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     3601 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     3661 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     3721 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     3781 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     3841 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     3901 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     3961 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     4021 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     4081 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     4141 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     4201 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     4261 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     4321 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     4381 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     4441 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     4501 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     4561 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     4621 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     4681 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     4741 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     4801 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     4861 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     4921 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     4981 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     5041 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     5101 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     5161 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     5221 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     5281 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     5341 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     5401 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     5461 acctaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc
     5521 tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc
     5581 gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac
     5641 tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca
     5701 ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg
     5761 agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag
     5821 aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc
     5881 accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt cccattcgcc attcaggctg
     5941 cgcaactgtt g
//
