LOCUS       sequence-243609-        5597 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of Human TRIM28 with deleted RING, Bbox, CC 
            domains.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5597)
  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 5597)
  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-TRIM28deltaRBCC
FEATURES             Location/Qualifiers
     source          1..5597
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        42..1019
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     promoter        1054..1072
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1054..1071
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             1107..1187
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAYPYDVPDYAYPYDVPDYA"
     polyA_signal    2668..2802
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2692..2711)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2746..2765
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2915..2935)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2915..2933)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2954..2970)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2954..2970)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2967..2989)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2978..2994
                     /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(3002..3008,3009..3026,3027..3032))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3047..3068
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3185..3202)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3356..3944)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3436..3455)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4115..4906,4907..4975))
                     /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     4738..4757
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4976..5080)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5106..5561)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5243..5264)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5455..5474
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 ggcgatcggt gcgggcctct tcgctattac gccagtcgac cgccaattca atatggcgta
       61 tatggactca tgccaattca atatggtgga tctggacctg tgccaattca atatggcgta
      121 tatggactcg tgccaattca atatggtgga tctggacccc agccaattca atatggcgga
      181 cttggcacca tgccaattca atatggcgga cctggcactg tgccaactgg ggaggggtct
      241 acttggcacg gtgccaagtt tgaggagggg tcttggccct gtgccaagtc cgccatattg
      301 aattggcatg gtgccaataa tggcggccat attggctata tgccaggatc aatatatagg
      361 caatatccaa tatggcccta tgccaatatg gctattggcc aggttcaata ctatgtattg
      421 gccctatgcc atatagtatt ccatatatgg gttttcctat tgacgtagat agcccctccc
      481 aatgggcggt cccatatacc atatatgggg cttcctaata ccgcccatag ccactccccc
      541 attgacgtca atggtctcta tatatggtct ttcctattga cgtcatatgg gcggtcctat
      601 tgacgtatat ggcgcctccc ccattgacgt caattacggt aaatggcccg cctggctcaa
      661 tgcccattga cgtcaatagg accacccacc attgacgtca atgggatggc tcattgccca
      721 ttcatatccg ttctcacgcc ccctattgac gtcaatgacg gtaaatggcc cacttggcag
      781 tacatcaata tctattaata gtaacttggc aagtacatta ctattggaag tacgccaggg
      841 tacattggca gtactcccat tgacgtcaat ggcggtaaat ggcccgcgat ggctgccaag
      901 tacatcccca ttgacgtcaa tggggagggg caatgacgca aatgggcgtt ccattgacgt
      961 aaatgggcgg taggcgtgcc taatgggagg tctatataag caatgctcgt ttagggaacc
     1021 gccattctgc ctggggacgt cggagcaagc ttgatttagg tgacactata gaatacaagc
     1081 tacttgttct ttttgcagga tccatgtacc catacgatgt tccagattac gcttatcctt
     1141 atgacgtacc tgactatgca tacccttatg atgtaccaga ctacgctggc cggccaatgg
     1201 cggcctccgc ggcggcagcc tcggcagcag cggcctcggc cgcctctggc agcatgattg
     1261 tggatcccgt ggagccacat ggcgagatga agtttcagtg ggacctcaat gcctggacca
     1321 agagtgccga ggcctttggc aagattgtgg cagagcgtcc tggcactaac tcaacaggcc
     1381 ctgcacccat ggcccctcca agagccccag ggcccctgag caagcagggc tctggcagca
     1441 gccagcccat ggaggtgcag gaaggctatg gctttgggtc aggagatgat ccctactcaa
     1501 gtgcagagcc ccatgtgtca ggtgtgaaac ggtcccgctc aggtgagggc gaggtgagcg
     1561 gccttatgcg caaggtgcca cgagtgagcc ttgaacgcct ggacctggac ctcacagctg
     1621 acagccagcc acccgtcttc aaggtcttcc caggcagtac cactgaggac tacaacctta
     1681 ttgttattga acgtggcgct gccgctgcag ctaccggcca gccagggact gcgcctgcag
     1741 gaacccctgg tgccccaccc ctggctggca tggccattgt caaggaggag gagacggagg
     1801 ctgccattgg agcccctcct actgccactg agggccctga gaccaaacct gtgcttatgg
     1861 ctcttgcgga gggtcctggt gctgagggtc cccgcctggc ctcacctagt ggcagcacca
     1921 gctcagggct ggaggtggtg gctcctgagg gtacctcagc cccaggtggt ggcccgggaa
     1981 ccctggatga cagtgccacc atttgccgtg tctgccagaa gccaggcgat ctggttatgt
     2041 gcaaccagtg tgagttttgt ttccacctgg actgtcacct gccggccctg caggatgtac
     2101 caggggagga gtggagctgc tcactctgcc atgtgctccc tgacctgaag gaggaggatg
     2161 gcagcctcag cctggatggt gcagacagca ctggcgtggt ggccaagctc tcaccagcca
     2221 accagcggaa atgtgagcgt gtactgctgg ccctattctg tcacgaaccc tgccgccccc
     2281 tgcatcagct ggctaccgac tccaccttct ccctggacca gcccggtggc accctggatc
     2341 tgaccctgat ccgtgcccgc ctccaggaga agttgtcacc tccctacagc tccccacagg
     2401 agtttgccca ggatgtgggc cgcatgttca agcaattcaa caagttaact gaggacaagg
     2461 cagacgtgca gtccatcatc ggcctgcagc gcttcttcga gacgcgcatg aacgaggcct
     2521 tcggtgacac caagttctct gctgtgctgg tggagccccc gccgatgagc ctgcctggtg
     2581 ctggcctgag ttcccaggag ctgtctggtg gccctggtga tggcccctga ggcgcgcctc
     2641 tagaactata gtgagtcgta ttacgtagat ccagacatga taagatacat tgatgagttt
     2701 ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct
     2761 attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt
     2821 cattttatgt ttcaggttca gggggaggtg tgggaggttt tttaattcgc ggccgcggcg
     2881 ccaatgcatt gggcccggta cccagctttt gttcccttta gtgagggtta attgcgcgct
     2941 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     3001 acaacatacg agccgggagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     3061 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     3121 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     3181 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     3241 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     3301 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     3361 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     3421 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     3481 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     3541 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     3601 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     3661 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     3721 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     3781 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     3841 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     3901 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     3961 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     4021 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     4081 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     4141 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     4201 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     4261 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     4321 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     4381 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     4441 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     4501 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     4561 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     4621 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     4681 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     4741 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     4801 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     4861 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     4921 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     4981 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     5041 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     5101 cacctaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag
     5161 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac
     5221 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga
     5281 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc
     5341 accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
     5401 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
     5461 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
     5521 caccacaccc gccgcgctta atgcgccgct acagggcgcg tcccattcgc cattcaggct
     5581 gcgcaactgt tgggaag
//
