LOCUS       sequence-243627-        6270 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of Human TRIM28 with deleted CC domain.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6270)
  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 6270)
  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-TRIM28deltaCC1
FEATURES             Location/Qualifiers
     source          1..6270
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        65..1049
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     promoter        1084..1102
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1084..1101
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             1137..1217
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAYPYDVPDYAYPYDVPDYA"
     polyA_signal    3370..3504
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3394..3413)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3448..3467
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3617..3637)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3617..3635)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3656..3672)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3656..3672)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3669..3691)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3680..3696
                     /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(3704..3710,3711..3728,3729..3734))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3749..3770
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3887..3904)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4058..4647)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4138..4157)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4818..5609,5610..5678))
                     /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     5441..5460
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5679..5783)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5809..6264)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5946..5967)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6158..6177
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac
       61 gccagtcgac cgccaattca atatggcgta tatggactca tgccaattca atatggtgga
      121 tctggacctg tgccaattca atatggcgta tatggactcg tgccaattca atatggtgga
      181 tctggacccc agccaattca atatggcgga cttggcacca tgccaattca atatggcgga
      241 cctggcactg tgccaactgg ggaggggtct acttggcacg gtgccaagtt tgaggagggg
      301 tcttggccct gtgccaagtc cgccatattg aattggcatg gtgccaataa tggcggccat
      361 attggctata tgccaggatc aatatatagg caatatccaa tatggcccta tgccaatatg
      421 gctattggcc aggttcaata ctatgtattg gccctatgcc atatagtatt ccatatatgg
      481 gttttcctat tgacgtagat agcccctccc aatgggcggt cccatatacc atatatgggg
      541 cttcctaata ccgcccatag ccactccccc attgacgtca atggtctcta tatatggtct
      601 ttcctattga cgtcatatgg gcggtcctat tgacgtatat ggcgcctccc ccattgacgt
      661 caattacggt aaatggcccg cctggctcaa tgcccattga cgtcaatagg accacccacc
      721 attgacgtca atgggatggc tcattgccca ttcatatccg ttctcacgcc ccctattgac
      781 gtcaatgacg gtaaatggcc cacttggcag tacatcaata tctattaata gtaacttggc
      841 aagtacatta ctattggaag tacgccaggg tacattggca gtactcccat tgacgtcaat
      901 ggcggtaaat ggcccgcgat ggctgccaag tacatcccca ttgacgtcaa tggggagggg
      961 caatgacgca aatgggcgtt ccattgacgt aaatgggcgg taggcgtgcc taatgggagg
     1021 tctatataag caatgctcgt ttagggaacc gccattctgc ctggggacgt cggagcaagc
     1081 ttgatttagg tgacactata gaatacaagc tacttgttct ttttgcagga tccatgtacc
     1141 catacgatgt tccagattac gcttatcctt atgacgtacc tgactatgca tacccttatg
     1201 atgtaccaga ctacgctggc cggccaatgg cggcctccgc ggcggcagcc tcggcagcag
     1261 cggcctcggc cgcctctggc agcccgggcc cgggcgaggg ctccgctggc ggcgaaaagc
     1321 gctccaccgc cccttcggcc gcagcctcgg cctctgcctc agccgcggcg tcgtcgcccg
     1381 cggggggcgg cgccgaggcg ctggagctgc tggagcactg cggcgtgtgc agagagcgcc
     1441 tgcgacccga gagggagccc cgcctgctgc cctgtttgca ctcggcctgt agtgcctgct
     1501 tagggcccgc ggcccccgcc gccgccaaca gctcggggga cggcggggcg gcgggcgacg
     1561 gcaccgtggt ggactgtccc gtgtgcaagc aacagtgctt ctccaaagac atcgtggaga
     1621 attatttcat gcgtgatagt ggcagcaagg ctgccaccga cgcccaggat gcgaaccagt
     1681 gctgcactag ctgtgaggat aatgccccag ccaccagcta ctgtgtggag tgctcggagc
     1741 ctctgtgtga gacctgtgta gaggcgcacc agcgggtgaa gtacaccaag gaccatactg
     1801 tgcgctctac tgggccagcc aagtctcggg atggtgaacg tactgtctat tgcaacgtac
     1861 acaagcatga accccttgtg ctgttttgtg agagctgtga tactctcacc tgccgagact
     1921 gccagctcaa tgcccacaag gaccaccagt accagatgat tgtggatccc gtggagccac
     1981 atggcgagat gaagtttcag tgggacctca atgcctggac caagagtgcc gaggcctttg
     2041 gcaagattgt ggcagagcgt cctggcacta actcaacagg ccctgcaccc atggcccctc
     2101 caagagcccc agggcccctg agcaagcagg gctctggcag cagccagccc atggaggtgc
     2161 aggaaggcta tggctttggg tcaggagatg atccctactc aagtgcagag ccccatgtgt
     2221 caggtgtgaa acggtcccgc tcaggtgagg gcgaggtgag cggccttatg cgcaaggtgc
     2281 cacgagtgag ccttgaacgc ctggacctgg acctcacagc tgacagccag ccacccgtct
     2341 tcaaggtctt cccaggcagt accactgagg actacaacct tattgttatt gaacgtggcg
     2401 ctgccgctgc agctaccggc cagccaggga ctgcgcctgc aggaacccct ggtgccccac
     2461 ccctggctgg catggccatt gtcaaggagg aggagacgga ggctgccatt ggagcccctc
     2521 ctactgccac tgagggccct gagaccaaac ctgtgcttat ggctcttgcg gagggtcctg
     2581 gtgctgaggg tccccgcctg gcctcaccta gtggcagcac cagctcaggg ctggaggtgg
     2641 tggctcctga gggtacctca gccccaggtg gtggcccggg aaccctggat gacagtgcca
     2701 ccatttgccg tgtctgccag aagccaggcg atctggttat gtgcaaccag tgtgagtttt
     2761 gtttccacct ggactgtcac ctgccggccc tgcaggatgt accaggggag gagtggagct
     2821 gctcactctg ccatgtgctc cctgacctga aggaggagga tggcagcctc agcctggatg
     2881 gtgcagacag cactggcgtg gtggccaagc tctcaccagc caaccagcgg aaatgtgagc
     2941 gtgtactgct ggccctattc tgtcacgaac cctgccgcct cctgcatcag ctggctaccg
     3001 actccacctt ctccctggac cagcccggtg gcaccctgga tctgaccctg atccgtgccc
     3061 gcctccagga gaagttgtca cctccctaca gctccccaca ggagtttgcc caggatgtgg
     3121 gccgcatgtt caagcaattc aacaagttaa ctgaggacaa ggcagacgtg cagtccatca
     3181 tcggcctgca gcgcttcttc gagacgcgca tgaacgaggc cttcggtgac accaagttct
     3241 ctgctgtgct ggtggagccc ccgccgatga gcctgcctgg tgctggcctg agttcccagg
     3301 agctgtctgg tggccctggt gatggcccct gaggcgcgcc tctagaacta tagtgagtcg
     3361 tattacgtag atccagacat gataagatac attgatgagt ttggacaaac cacaactaga
     3421 atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc
     3481 attataagct gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt
     3541 cagggggagg tgtgggaggt tttttaattc gcggccgcgg cgccaatgca ttgggcccgg
     3601 tacccagctt ttgttccctt tagtgagggt taattgcgcg cttggcgtaa tcatggtcat
     3661 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggga
     3721 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     3781 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     3841 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     3901 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     3961 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     4021 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     4081 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     4141 gataccaggc gtttcccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     4201 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     4261 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     4321 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     4381 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     4441 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     4501 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     4561 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     4621 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     4681 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     4741 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     4801 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     4861 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     4921 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     4981 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     5041 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     5101 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     5161 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     5221 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     5281 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     5341 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     5401 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     5461 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     5521 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     5581 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     5641 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     5701 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     5761 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctaa attgtaagcg
     5821 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat
     5881 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg
     5941 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc
     6001 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt
     6061 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag
     6121 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg
     6181 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc
     6241 ttaatgcgcc gctacagggc gcgtcccatt
//
