LOCUS       sequence-193627-        6247 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of DNAJB1(exon1)-PRKACA(exon2-Cterminus) 
            chimeric protein from fibrolamellar hepatocellular carcinoma.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6247)
  AUTHORS   Honeyman JN, Simon EP, Robine N, Chiaroni-Clarke R, Darcy DG, Lim 
            II, Gleason CE, Murphy JM, Rosenberg BR, Teegan L, Takacs CN, Botero
            S, Belote R, Germer S, Emde AK, Vacic V, Bhanot U, LaQuaglia MP, 
            Simon SM
  TITLE     Detection of a recurrent DNAJB1-PRKACA chimeric transcript in 
            fibrolamellar hepatocellular carcinoma.
  JOURNAL   Science. 2014 Feb 28;343(6174):1010-4. doi: 10.1126/science.1249484.
  PUBMED    24578576
REFERENCE   2  (bases 1 to 6247)
  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: "10.1126/science"; 
            journalName: "Science"; date: "2014-02-28- 28"; volume: "343"; 
            issue: "6174"; pages: "1010-4"
COMMENT     Sequence Label: pcrDNA3.1-Chimera
FEATURES             Location/Qualifiers
     source          1..6247
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        10..389
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        390..593
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     544..564
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     638..657
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        638..656
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      713..722
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     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      complement(2356..2944)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2436..2455)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3273..3320
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     3345..3364
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(3552..4346)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             complement(3552..4346)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3663..3682)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4273..4292
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(4381..4738)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4395..4530
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        4547..4738
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     4695..4715
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     CDS             complement(join(4798..5589,5590..5658))
                     /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     5421..5440
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5659..5763)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5790..6245
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5877..5896)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6087..6108
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 gcgcgcgttg acattgatta ttgactagtt attaatagta atcaattacg gggtcattag
       61 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
      121 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
      181 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
      241 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
      301 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
      361 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
      421 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
      481 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
      541 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctctctggc
      601 taactagaga acccactgct tactggctta tcgaaattaa tacgactcac tatagggaga
      661 cccaagctgg ctagcgttta aacttaagct tggtaccgag ctcggatccg ccgccaccat
      721 gggtaaagac tactaccaga cgttgggcct ggcccgcggc gcgtcggacg aggagatcaa
      781 gcgggcctac cgccgccagg cgctgcgcta ccacccggac aagaacaagg agcccggcgc
      841 cgaggagaag ttcaaggaga tcgctgaggc ctacgacgtg ctcagcgacc cgcgcaagcg
      901 cgagatcttc gaccgctacg gggaggaagt gaaagaattc ttagccaaag ccaaagaaga
      961 ttttcttaaa aaatgggaaa gtcccgctca gaacacagcc cacttggatc agtttgaacg
     1021 aatcaagacc ctcggcacgg gctccttcgg gcgggtgatg ctggtgaaac acaaggagac
     1081 cgggaaccac tatgccatga agatcctcga caaacagaag gtggtgaaac tgaaacagat
     1141 cgaacacacc ctgaatgaaa agcgcatcct gcaagctgtc aactttccgt tcctcgtcaa
     1201 actcgagttc tccttcaagg acaactcaaa cttatacatg gtcatggagt acgtgcccgg
     1261 cggggagatg ttctcacacc tacggcggat cggaaggttc agtgagcccc atgcccgttt
     1321 ctacgcggcc cagatcgtcc tgacctttga gtatctgcac tcgctggatc tcatctacag
     1381 ggacctgaag ccggagaatc tgctcattga ccagcagggc tacattcagg tgacagactt
     1441 cggtttcgcc aagcgcgtga agggccgcac ttggaccttg tgcggcaccc ctgagtacct
     1501 ggcccctgag attatcctga gcaaaggcta caacaaggcc gtggactggt gggccctggg
     1561 ggttcttatc tatgaaatgg ccgctggcta cccgcccttc ttcgcagacc agcccatcca
     1621 gatctatgag aagatcgtct ctgggaaggt gcgcttccct tcccacttca gctctgactt
     1681 gaaggacctg ctgcggaacc tcctgcaggt agatctcacc aagcgctttg ggaacctcaa
     1741 gaatggggtc aacgatatca agaaccacaa gtggtttgcc acaactgact ggattgccat
     1801 ctaccagagg aaggtggaag ctcccttcat accaaagttt aaaggccctg gggatacgag
     1861 taactttgac gactatgagg aagaagaaat ccgggtctcc atcaatgaga agtgtggcaa
     1921 ggagttttct gagttttagt aatgattaag cggccgctcg agtctagagg gcccgtttaa
     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 aaccagtggc ggtaatacgg ttatccacag aatcagggga
     2281 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     2341 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     2401 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     2461 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     2521 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     2581 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     2641 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     2701 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     2761 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     2821 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     2881 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     2941 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     3001 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     3061 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa cctgaggcta tggcagggcc
     3121 tgccgccccg acgttggctg cgagccctgg gccttcaccc gaacttgggg ggtggggtgg
     3181 ggaaaaggaa gaaacgcggg cgtattggcc ccaatggggt ctcggtgggg tatcgacaga
     3241 gtgccagccc tgggaccgaa ccccgcgttt atgaacaaac gacccaacac cgtgcgtttt
     3301 attctgtctt tttattgccg tcatagcgcg ggttccttcc ggtattgtct ccttccgtgt
     3361 ttcagttagc ctccccctag ggtgggcgaa gaactccagc atgagatccc cgcgctggag
     3421 gatcatccag ccggcgtccc ggaaaacgat tccgaagccc aacctttcat agaaggcggc
     3481 ggtggaatcg aaatctcgtg atggcaggtt gggcgtcgct tggtcggtca tttcgaaccc
     3541 cagagtcccg ctcagaagaa ctcgtcaaga aggcgataga aggcgatgcg ctgcgaatcg
     3601 ggagcggcga taccgtaaag cacgaggaag cggtcagccc attcgccgcc aagctcttca
     3661 gcaatatcac gggtagccaa cgctatgtcc tgatagcggt ccgccacacc cagccggcca
     3721 cagtcgatga atccagaaaa gcggccattt tccaccatga tattcggcaa gcaggcatcg
     3781 ccatgggtca cgacgagatc ctcgccgtcg ggcatgctcg ccttgagcct ggcgaacagt
     3841 tcggctggcg cgagcccctg atgctcttcg tccagatcat cctgatcgac aagaccggct
     3901 tccatccgag tacgtgctcg ctcgatgcga tgtttcgctt ggtggtcgaa tgggcaggta
     3961 gccggatcaa gcgtatgcag ccgccgcatt gcatcagcca tgatggatac tttctcggca
     4021 ggagcaaggt gagatgacag gagatcctgc cccggcactt cgcccaatag cagccagtcc
     4081 cttcccgctt cagtgacaac gtcgagcaca gctgcgcaag gaacgcccgt cgtggccagc
     4141 cacgatagcc gcgctgcctc gtcttgcagt tcattcaggg caccggacag gtcggtcttg
     4201 acaaaaagaa ccgggcgccc ctgcgctgac agccggaaca cggcggcatc agagcagccg
     4261 attgtctgtt gtgcccagtc atagccgaat agcctctcca cccaagcggc cggagaacct
     4321 gcgtgcaatc catcttgttc aatcatgcga aacgatcctc atcctgtctc ttgatcgatc
     4381 tttgcaaaag cctaggcctc caaaaaagcc tcctcactac ttctggaata gctcagaggc
     4441 cgaggcggcc tcggcctctg cataaataaa aaaaattagt cagccatggg gcggagaatg
     4501 ggcggaactg ggcggagtta ggggcgggat gggcggagtt aggggcggga ctatggttgc
     4561 tgactaattg agatgcatgc tttgcatact tctgcctgct ggggagcctg gggactttcc
     4621 acacctggtt gctgactaat tgagatgcat gctttgcata cttctgcctg ctggggagcc
     4681 tggggacttt ccacacccta actgacacac attccacagc tggttctttc cgcctcagga
     4741 ctcttccttt ttcaataaat caatctaaag tatatatgag taaacttggt ctgacagtta
     4801 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     4861 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     4921 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     4981 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     5041 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     5101 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     5161 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     5221 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     5281 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     5341 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     5401 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     5461 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     5521 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     5581 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     5641 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     5701 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     5761 gcgcacattt ccccgaaaag tgccacctga cgcgccctgt agcggcgcat taagcgcggc
     5821 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     5881 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     5941 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     6001 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     6061 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     6121 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     6181 aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac
     6241 aatttac
//
