LOCUS       sequence-160532-        7274 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  EZH2 minigene to test splicing event.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7274)
  AUTHORS   Kim E, Ilagan JO, Liang Y, Daubner GM, Lee SC, Ramakrishnan A, Li Y,
            Chung YR, Micol JB, Murphy ME, Cho H, Kim MK, Zebari AS, Aumann S, 
            Park CY, Buonamici S, Smith PG, Deeg HJ, Lobry C, Aifantis I, Modis 
            Y, Allain FH, Halene S, Bradley RK, Abdel-Wahab O
  TITLE     SRSF2 Mutations Contribute to Myelodysplasia by Mutant-Specific 
            Effects on Exon Recognition.
  JOURNAL   Cancer Cell. 2015 May 11;27(5):617-30. doi: 
            10.1016/j.ccell.2015.04.006.
  PUBMED    25965569
REFERENCE   2  (bases 1 to 7274)
  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: 
            "10.1016/j.ccell.2015.04"; journalName: "Cancer Cell"; date: 
            "2015-05-11- 11"; volume: "27"; issue: "5"; pages: "617-30"
COMMENT     Sequence Label: pUB6_EZH2
FEATURES             Location/Qualifiers
     source          1..7274
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        18..1227
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     1064..1083
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             complement(2060..2071)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             3138..3179
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3189..3206
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3229..3246)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3235..3459
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3505..3933
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3592..3611)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3802..3823
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3942..3962)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3947..4277
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4128..4263
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4190..4209
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4325..4372
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4391..4789
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    4947..5068
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4984..5003)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5038..5057
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5117..5133)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5117..5133)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5130..5152)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5141..5157
                     /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(5165..5171,5172..5189,5190..5195))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5210..5231
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5348..5365)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5519..6107)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5599..5618)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6278..7069,7070..7138))
                     /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     6901..6920
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7139..7243)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctggc ctccgcgccg ggttttggcg cctcccgcgg gcgcccccct
       61 cctcacggcg agcgctgcca cgtcagacga agggcgcagg agcgtcctga tccttccgcc
      121 cggacgctca ggacagcggc ccgctgctca taagactcgg ccttagaacc ccagtatcag
      181 cagaaggaca ttttaggacg ggacttgggt gactctaggg cactggtttt ctttccagag
      241 agcggaacag gcgaggaaaa gtagtccctt ctcggcgatt ctgcggaggg atctccgtgg
      301 ggcggtgaac gccgatgatt atataaggac gcgccgggtg tggcacagct agttccgtcg
      361 cagccgggat ttgggtcgcg gttcttgttt gtggatcgct gtgatcgtca cttggtgagt
      421 agcgggctgc tgggctggcc ggggctttcg tggccgccgg gccgctcggt gggacggaag
      481 cgtgtggaga gaccgccaag ggctgtagtc tgggtccgcg agcaaggttg ccctgaactg
      541 ggggttgggg ggagcgcagc aaaatggcgg ctgttcccga gtcttgaatg gaagacgctt
      601 gtgaggcggg ctgtgaggtc gttgaaacaa ggtggggggc atggtgggcg gcaagaaccc
      661 aaggtcttga ggccttcgct aatgcgggaa agctcttatt cgggtgagat gggctggggc
      721 accatctggg gaccctgacg tgaagtttgt cactgactgg agaactcggt ttgtcgtctg
      781 ttgcgggggc ggcagttatg cggtgccgtt gggcagtgca cccgtacctt tgggagcgcg
      841 cgccctcgtc gtgtcgtgac gtcacccgtt ctgttggctt ataatgcagg gtggggccac
      901 ctgccggtag gtgtgcggta ggcttttctc cgtcgcagga cgcagggttc gggcctaggg
      961 taggctctcc tgaatcgaca ggcgccggac ctctggtgag gggagggata agtgaggcgt
     1021 cagtttcttt ggtcggtttt atgtacctat cttcttaagt agctgaagct ccggttttga
     1081 actatgcgct cggggttggc gagtgtgttt tgtgaagttt tttaggcacc ttttgaaatg
     1141 taatcatttg ggtcaatatg taattttcag tgttagacta gtaaattgtc cgctaaattc
     1201 tggccgtttt tggctttttt gttagacgaa gcttggtacc gagctcggat ccactagtcc
     1261 agtgtggtgg aattctgcag atcttttcat gcaacaccca acacttataa gcggaagaac
     1321 acagaaacag ctctagacaa caaaccttgt ggaccacagt gttaccagca tttggtaaga
     1381 cttagtgcct aattattgca gagggtactt gagaggactt tgcactttgg tggaggtgat
     1441 caagtcagcg ttaatgttgt ctgcacccat gctgttatgg gccatatgat agacagagct
     1501 ttctcaccca tgctgtttct tccatagctc atgtactgca cattttagtc cttcattaat
     1561 gtggtattat tctttttaaa atctgtgtta tctcttcagc aacatttcaa ggtctttagg
     1621 atcaaggact tttatgcctt ccacaaagtc tatgagagtt gcaaggcaca gatagtccgc
     1681 ttagaaagca ggcatgcagt aatcatacgc agtagatccc aagcccctct catagccata
     1741 tccctctact tgagttttag atgcactgtc atttcagcta tgtgataggt tccacagatt
     1801 tttttttttt aaatgatgga gaacaggtaa ggggttagga aaatatgagg tagaaaagtt
     1861 gtttttacgg gtactctctt atttttacca tttcccctct ttgaaccaag ccactcctac
     1921 ctaggaacta aatgggtata tattgcctgt tggattttgg acagaagatt cattgaatgg
     1981 cacctgcaga aggtatccta tttttaagca atttggtttt tgtagaatga agtataaaac
     2041 tgttggctgc tgtcaaactt ctcccttcta tttccactaa aaaaatggaa ttcttgagca
     2101 cttgttttat aatattcttt acaaggcaga attttgtttg gctcatctaa aatgtttctg
     2161 tggtctctat aaaccatgct cttaccattg ttttatactt tgagactgga ttattgtact
     2221 tcataatgca agatgtaacc ctcacagtct tattcctgaa gttcctttca tgtcttcctg
     2281 tggctgtgta tccttcacag gaaaaaaaaa aatgttggat gttttaatag taagacagga
     2341 agtgacaaag tcaagcatgt ttgtgttttg tgttcttgta tagttgaaat attaacacga
     2401 agaggtatgc attatcttaa actctgcata ggtagtctga ctccgtgcac attaggtata
     2461 tgattctttt gtaaagattc acttttaaat agactgtatg tccatagaga taattacctt
     2521 tattacatga agtatatttc atactttatc ttcttatatt ttaacttaat tataactagc
     2581 taatatattt tacaaattca caatatagaa ctgtcttgcc tgatttttgt accattctag
     2641 gaatatatag aacccatatt tctctttctg aaattttcag actatcaaaa gcatatcaaa
     2701 acaagtatat tagacatgca aatgattatt tgtgataaat ggataatgtg atacattttt
     2761 tgactaacct ggcttatata gtattttttt ttctcttcca tcaaaatgag ttttagaact
     2821 ttgccctgat gttgacattt ttcatttcgt aggagggagc aaaggagttt gctgctgctc
     2881 tcaccgctga gcggataaag accccaccaa aacgtccagg aggccgcaga agaggacggc
     2941 ttcccaataa cagtagcagg cccagcaccc ccaccattaa tgtgctggaa tcaaaggata
     3001 cagacagtga tagggaagca gggactgaaa cggggggaga gaacaatgat aaagaagaag
     3061 aagagaagaa agatgaaact tcgagctcct ctgatccagc acagtggcgg ccgctcgagt
     3121 ctagagggcc cttcgaaggt aagcctatcc ctaaccctct cctcggtctc gattctacgc
     3181 gtaccggtca tcatcaccat caccattgag tttaaacccg ctgatcagcc tcgactgtgc
     3241 cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag
     3301 gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta
     3361 ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag
     3421 acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca
     3481 gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     3541 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     3601 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     3661 gcatcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     3721 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     3781 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     3841 tctcggtcta ttcttttgat ttataaggga ttttggggat ttcggcctat tggttaaaaa
     3901 atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg
     3961 gtgtggaaag tccccaggct ccccaggcag gcagaagtat gcaaagcatg catctcaatt
     4021 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     4081 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     4141 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     4201 aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     4261 gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcagc
     4321 acgtgttgac aattaatcat cggcatagta tatcggcata gtataatacg acaaggtgag
     4381 gaactaaacc atggccaagc ctttgtctca agaagaatcc accctcattg aaagagcaac
     4441 ggctacaatc aacagcatcc ccatctctga agactacagc gtcgccagcg cagctctctc
     4501 tagcgacggc cgcatcttca ctggtgtcaa tgtatatcat tttactgggg gaccttgtgc
     4561 agaactcgtg gtgctgggca ctgctgctgc tgcggcagct ggcaacctga cttgtatcgt
     4621 cgcgatcgga aatgagaaca ggggcatctt gagcccctgc ggacggtgcc gacaggtgct
     4681 tctcgatctg catcctggga tcaaagccat agtgaaggac agtgatggac agccgacggc
     4741 agttgggatt cgtgaattgc tgccctctgg ttatgtgtgg gagggctaag cacttcgtgg
     4801 ccgaggagca ggactgacac gtgctacgag atttcgattc caccgccgcc ttctatgaaa
     4861 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
     4921 tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat
     4981 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     5041 gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga
     5101 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     5161 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     5221 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     5281 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     5341 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     5401 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     5461 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     5521 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     5581 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     5641 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     5701 tggcgctttc tcaatgctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     5761 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     5821 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     5881 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     5941 actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct
     6001 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     6061 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     6121 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     6181 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     6241 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     6301 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     6361 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     6421 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     6481 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     6541 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     6601 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     6661 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     6721 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     6781 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     6841 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     6901 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     6961 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     7021 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     7081 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     7141 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     7201 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     7261 tgccacctga cgtc
//
