LOCUS       sequence-160535-        6039 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  EPB49 minigene to test splicing event.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6039)
  AUTHORS   Ilagan JO, Ramakrishnan A, Hayes B, Murphy ME, Zebari AS, Bradley P,
            Bradley RK
  TITLE     U2AF1 mutations alter splice site recognition in hematological 
            malignancies.
  JOURNAL   Genome Res. 2015 Jan;25(1):14-26. doi: 10.1101/gr.181016.114. Epub 
            2014 Sep 29.
  PUBMED    25267526
REFERENCE   2  (bases 1 to 6039)
  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: 
            "10.1101/gr.181016.114"; journalName: "Genome Res"; date: "2015-01";
            volume: "25"; issue: "1"; pages: "14-26"
COMMENT     Sequence Label: pUB6_EPB49
FEATURES             Location/Qualifiers
     source          1..6039
                     /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             1903..1944
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             1954..1971
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1994..2011)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2000..2224
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2270..2698
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2357..2376)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2567..2588
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2707..2727)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2712..3042
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2893..3028
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2955..2974
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3090..3137
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3156..3554
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    3712..3833
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3749..3768)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3803..3822
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3882..3898)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3882..3898)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3895..3917)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3906..3922
                     /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(3930..3936,3937..3954,3955..3960))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3975..3996
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4113..4130)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4284..4872)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4364..4383)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5043..5834,5835..5903))
                     /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     5666..5685
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5904..6008)
                     /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 atgcctgcag gtgagtgcct cctggagggg aacaggcaga
     1321 gcagcagaag ctgggtcctg gcacccgaaa gtgtcagggg aaggggggtg tggggccaca
     1381 gagttgcctg gtgaagaaga agaagccggg gtaaggctgg gaaggtctcc ctgccccgct
     1441 ctggctcact gcggctttgg tctccccaga acggagaggg ccagaggggg aggatggacc
     1501 gggggaactc cctgccctgt gtgctggagc agaaggtgag gggcagggga cacaagcgcc
     1561 tgtagcgggg cgggaggctg gggagatgcc aggcaagtgg aatgtgctgc ggggtcttcc
     1621 tggtgttgaa gatgggaggg atggcaggga agttggggga gacagagggc aagatggcat
     1681 gaaccccagt ggcctctgca ggttgatgtg ggaggcctag ggagtcgaag gacaggatat
     1741 tctgtgagcc taagattgcc cctcccccca cccccagatc tatccctatg aaatgctagt
     1801 ggtgaccaac aaggggcgaa ccaagctgcc accgggggtg gatcggatgc ggcttgagat
     1861 ccagcacagt ggcggccgct cgagtctaga gggcccttcg aaggtaagcc tatccctaac
     1921 cctctcctcg gtctcgattc tacgcgtacc ggtcatcatc accatcacca ttgagtttaa
     1981 acccgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc
     2041 cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag
     2101 gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag
     2161 gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct
     2221 atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt
     2281 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     2341 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     2401 tttccccgtc aagctctaaa tcggggcatc cctttagggt tccgatttag tgctttacgg
     2461 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     2521 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     2581 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     2641 gggatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa
     2701 ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca ggcaggcaga
     2761 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     2821 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     2881 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     2941 tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag
     3001 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt
     3061 atatccattt tcggatctga tcagcacgtg ttgacaatta atcatcggca tagtatatcg
     3121 gcatagtata atacgacaag gtgaggaact aaaccatggc caagcctttg tctcaagaag
     3181 aatccaccct cattgaaaga gcaacggcta caatcaacag catccccatc tctgaagact
     3241 acagcgtcgc cagcgcagct ctctctagcg acggccgcat cttcactggt gtcaatgtat
     3301 atcattttac tgggggacct tgtgcagaac tcgtggtgct gggcactgct gctgctgcgg
     3361 cagctggcaa cctgacttgt atcgtcgcga tcggaaatga gaacaggggc atcttgagcc
     3421 cctgcggacg gtgccgacag gtgcttctcg atctgcatcc tgggatcaaa gccatagtga
     3481 aggacagtga tggacagccg acggcagttg ggattcgtga attgctgccc tctggttatg
     3541 tgtgggaggg ctaagcactt cgtggccgag gagcaggact gacacgtgct acgagatttc
     3601 gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc
     3661 tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt
     3721 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     3781 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
     3841 tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg
     3901 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     3961 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     4021 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     4081 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     4141 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     4201 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     4261 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     4321 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     4381 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     4441 tccgcctttc tcccttcggg aagcgtggcg ctttctcaat gctcacgctg taggtatctc
     4501 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     4561 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     4621 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     4681 acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc
     4741 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     4801 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     4861 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     4921 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     4981 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     5041 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     5101 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     5161 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     5221 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     5281 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     5341 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     5401 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     5461 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     5521 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     5581 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     5641 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     5701 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     5761 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     5821 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     5881 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     5941 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     6001 gttccgcgca catttccccg aaaagtgcca cctgacgtc
//
