LOCUS       sequence-243620-        6071 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 6071)
  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 6071)
  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-TRIM28deltaCC2
FEATURES             Location/Qualifiers
     source          1..6071
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        17..994
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     promoter        1029..1047
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1029..1046
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             1082..1162
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAYPYDVPDYAYPYDVPDYA"
     polyA_signal    3117..3251
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3141..3160)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3195..3214
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3364..3384)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3364..3382)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3403..3419)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3403..3419)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3416..3438)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3427..3443
                     /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(3451..3457,3458..3475,3476..3481))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3496..3517
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3634..3651)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3805..4393)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3885..3904)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4564..5355,5356..5424))
                     /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     5187..5206
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5425..5529)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5555..6010)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5692..5713)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5904..5923
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 attacgccag tcgaccgcca attcaatatg gcgtatatgg actcatgcca attcaatatg
       61 gtggatctgg acctgtgcca attcaatatg gcgtatatgg actcgtgcca attcaatatg
      121 gtggatctgg accccagcca attcaatatg gcggacttgg caccatgcca attcaatatg
      181 gcggacctgg cactgtgcca actggggagg ggtctacttg gcacggtgcc aagtttgagg
      241 aggggtcttg gccctgtgcc aagtccgcca tattgaattg gcatggtgcc aataatggcg
      301 gccatattgg ctatatgcca ggatcaatat ataggcaata tccaatatgg ccctatgcca
      361 atatggctat tggccaggtt caatactatg tattggccct atgccatata gtattccata
      421 tatgggtttt cctattgacg tagatagccc ctcccaatgg gcggtcccat ataccatata
      481 tggggcttcc taataccgcc catagccact cccccattga cgtcaatggt ctctatatat
      541 ggtctttcct attgacgtca tatgggcggt cctattgacg tatatggcgc ctcccccatt
      601 gacgtcaatt acggtaaatg gcccgcctgg ctcaatgccc attgacgtca ataggaccac
      661 ccaccattga cgtcaatggg atggctcatt gcccattcat atccgttctc acgcccccta
      721 ttgacgtcaa tgacggtaaa tggcccactt ggcagtacat caatatctat taatagtaac
      781 ttggcaagta cattactatt ggaagtacgc cagggtacat tggcagtact cccattgacg
      841 tcaatggcgg taaatggccc gcgatggctg ccaagtacat ccccattgac gtcaatgggg
      901 aggggcaatg acgcaaatgg gcgttccatt gacgtaaatg ggcggtaggc gtgcctaatg
      961 ggaggtctat ataagcaatg ctcgtttagg gaaccgccat tctgcctggg gacgtcggag
     1021 caagcttgat ttaggtgaca ctatagaata caagctactt gttctttttg caggatccat
     1081 gtacccatac gatgttccag attacgctta tccttatgac gtacctgact atgcataccc
     1141 ttatgatgta ccagactacg ctggccggcc aatggcggcc tccgcggcgg cagcctcggc
     1201 agcagcggcc tcggccgcct ctggcagccc gggcccgggc gagggctccg ctggcggcga
     1261 aaagcgctcc accgcccctt cggccgcagc ctcggcctct gcctcagccg cggcgtcgtc
     1321 gcccgcgggg ggcggcgccg aggcgctgga gctgctggag cactgcggcg tgtgcagaga
     1381 gcgcctgcga cccgagaggg agccccgcct gctgccctgt ttgcactcgg cctgtagtgc
     1441 ctgcttaggg cccgcggccc ccgccgccgc caacagctcg ggggacggcg gggcggcggg
     1501 cgacggcacc gtggtggact gtcccgtgtg caagcaacag tgcttctcca aagacatcgt
     1561 ggagaattat ttcatgcgtg atagtggcag caaggctgcc accgacgccc aggatgcgaa
     1621 ccagtgctgc actagctgtg aggataatgc cccagccacc agctactgtg tggagtgctc
     1681 ggagcctctg tgtgagacct gtgtagaggc gcaccagcgg gtgaagtaca ccaaggacca
     1741 tactgtgcgc tctactgggc cagccaagtc tcgggatggt gaacgtactg tctattgcaa
     1801 cgtacacaag catgaacccc ttgtgctgtt ttgtgagagc tgtgatactc tcacctgccg
     1861 agactgccag ctcaatgccc acaaggacca ccagtaccag gaggtgcagg aaggctatgg
     1921 ctttgggtca ggagatgatc cctactcaag tgcagagccc catgtgtcag gtgtgaaacg
     1981 gtcccgctca ggtgagggcg aggtgagcgg ccttatgcgc aaggtgccac gagtgagcct
     2041 tgaacgcctg gacctggacc tcacagctga cagccagcca cccgtcttca aggtcttccc
     2101 aggcagtacc actgaggact acaaccttat tgttattgaa cgtggcgctg ccgctgcagc
     2161 taccggccag ccagggactg cgcctgcagg aacccctggt gccccacccc tggctggcat
     2221 ggccattgtc aaggaggagg agacggaggc tgccattgga gcccctccta ctgccactga
     2281 gggccctgag accaaacctg tgcttatggc tcttgcggag ggtcctggtg ctgagggtcc
     2341 ccgcctggcc tcacctagtg gcagcaccag ctcagggctg gaggtggtgg ctcctgaggg
     2401 tacctcagcc ccaggtggtg gcccgggaac cctggatgac agtgccacca tttgccgtgt
     2461 ctgccagaag ccaggcgatc tggttatgtg caaccagtgt gagttttgtt tccacctgga
     2521 ctgtcacctg ccggccctgc aggatgtacc aggggaggag tggagctgct cactctgcca
     2581 tgtgctccct gacctgaagg aggaggatgg cagcctcagc ctggatggtg cagacagcac
     2641 tggcgtggtg gccaagctct caccagccaa ccagcggaaa tgtgagcgtg tactgctggc
     2701 cctattctgt cacgaaccct gccgccccct gcatcagctg gctaccgact ccaccttctc
     2761 cctggaccag cccggtggca ccctggatct gaccctgatc cgtgcccgcc tccaggagaa
     2821 gttgtcacct ccctacagct ccccacagga gtttgcccag gatgtgggcc gcatgttcaa
     2881 gcaattcaac aagttaactg aggacaaggc agacgtgcag tccatcatcg gcctgcagcg
     2941 cttcttcgag acgcgcatga acgaggcctt cggtgacacc aagttctctg ctgtgctggt
     3001 ggagcccccg ccgatgagcc tgcctggtgc tggcctgagt tcccaggagc tgtctggtgg
     3061 ccctggtgat ggcccctgag gcgcgcctct agaactatag tgagtcgtat tacgtagatc
     3121 cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa
     3181 aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca
     3241 ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag ggggaggtgt
     3301 gggaggtttt ttaattcgcg gccgcggcgc caatgcattg ggcccggtac ccagcttttg
     3361 ttccctttag tgagggttaa ttgcgcgctt ggcgtaatca tggtcatagc tgtttcctgt
     3421 gtgaaattgt tatccgctca caattccaca caacatacga gccgggagca taaagtgtaa
     3481 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     3541 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     3601 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     3661 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     3721 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     3781 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     3841 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     3901 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     3961 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     4021 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     4081 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     4141 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     4201 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     4261 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     4321 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     4381 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     4441 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     4501 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     4561 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     4621 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     4681 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     4741 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     4801 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     4861 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     4921 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     4981 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     5041 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     5101 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     5161 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     5221 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     5281 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     5341 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     5401 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     5461 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     5521 ggttccgcgc acatttcccc gaaaagtgcc acctaaattg taagcgttaa tattttgtta
     5581 aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc
     5641 aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg
     5701 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat
     5761 cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc
     5821 cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag
     5881 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg
     5941 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta
     6001 cagggcgcgt cccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg
     6061 gcctcttcgc t
//
