LOCUS       sequence-193622-        6082 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of PRKACA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6082)
  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 6082)
  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-PRKACA
FEATURES             Location/Qualifiers
     source          1..6082
                     /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(1829..1846)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1835..2059
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(2191..2779)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2271..2290)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3108..3155
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     3180..3199
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(3387..4181)
                     /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(3387..4181)
                     /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(3498..3517)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4108..4127
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(4216..4573)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4230..4365
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        4382..4573
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     4530..4550
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     CDS             complement(join(4633..5424,5425..5493))
                     /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     5256..5275
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5494..5598)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5625..6080
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5712..5731)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5922..5943
                     /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 gggcaacgcc gccgccgcca agaagggcag cgagcaggag agcgtgaaag aattcttagc
      781 caaagccaaa gaagattttc ttaaaaaatg ggaaagtccc gctcagaaca cagcccactt
      841 ggatcagttt gaacgaatca agaccctcgg cacgggctcc ttcgggcggg tgatgctggt
      901 gaaacacaag gagaccggga accactatgc catgaagatc ctcgacaaac agaaggtggt
      961 gaaactgaaa cagatcgaac acaccctgaa tgaaaagcgc atcctgcaag ctgtcaactt
     1021 tccgttcctc gtcaaactcg agttctcctt caaggacaac tcaaacttat acatggtcat
     1081 ggagtacgtg cccggcgggg agatgttctc acacctacgg cggatcggaa ggttcagtga
     1141 gccccatgcc cgtttctacg cggcccagat cgtcctgacc tttgagtatc tgcactcgct
     1201 ggatctcatc tacagggacc tgaagccgga gaatctgctc attgaccagc agggctacat
     1261 tcaggtgaca gacttcggtt tcgccaagcg cgtgaagggc cgcacttgga ccttgtgcgg
     1321 cacccctgag tacctggccc ctgagattat cctgagcaaa ggctacaaca aggccgtgga
     1381 ctggtgggcc ctgggggttc ttatctatga aatggccgct ggctacccgc ccttcttcgc
     1441 agaccagccc atccagatct atgagaagat cgtctctggg aaggtgcgct tcccttccca
     1501 cttcagctct gacttgaagg acctgctgcg gaacctcctg caggtagatc tcaccaagcg
     1561 ctttgggaac ctcaagaatg gggtcaacga tatcaagaac cacaagtggt ttgccacaac
     1621 tgactggatt gccatctacc agaggaaggt ggaagctccc ttcataccaa agtttaaagg
     1681 ccctggggat acgagtaact ttgacgacta tgaggaagaa gaaatccggg tctccatcaa
     1741 tgagaagtgt ggcaaggagt tttctgagtt ttagtaatga ttaagcggcc gctcgagtct
     1801 agagggcccg tttaaacccg ctgatcagcc tcgactgtgc cttctagttg ccagccatct
     1861 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt
     1921 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg
     1981 ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg
     2041 gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gtggcggtaa tacggttatc
     2101 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     2161 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     2221 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     2281 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     2341 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     2401 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     2461 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     2521 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     2581 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     2641 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     2701 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     2761 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     2821 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     2881 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaacctga
     2941 ggctatggca gggcctgccg ccccgacgtt ggctgcgagc cctgggcctt cacccgaact
     3001 tggggggtgg ggtggggaaa aggaagaaac gcgggcgtat tggccccaat ggggtctcgg
     3061 tggggtatcg acagagtgcc agccctggga ccgaaccccg cgtttatgaa caaacgaccc
     3121 aacaccgtgc gttttattct gtctttttat tgccgtcata gcgcgggttc cttccggtat
     3181 tgtctccttc cgtgtttcag ttagcctccc cctagggtgg gcgaagaact ccagcatgag
     3241 atccccgcgc tggaggatca tccagccggc gtcccggaaa acgattccga agcccaacct
     3301 ttcatagaag gcggcggtgg aatcgaaatc tcgtgatggc aggttgggcg tcgcttggtc
     3361 ggtcatttcg aaccccagag tcccgctcag aagaactcgt caagaaggcg atagaaggcg
     3421 atgcgctgcg aatcgggagc ggcgataccg taaagcacga ggaagcggtc agcccattcg
     3481 ccgccaagct cttcagcaat atcacgggta gccaacgcta tgtcctgata gcggtccgcc
     3541 acacccagcc ggccacagtc gatgaatcca gaaaagcggc cattttccac catgatattc
     3601 ggcaagcagg catcgccatg ggtcacgacg agatcctcgc cgtcgggcat gctcgccttg
     3661 agcctggcga acagttcggc tggcgcgagc ccctgatgct cttcgtccag atcatcctga
     3721 tcgacaagac cggcttccat ccgagtacgt gctcgctcga tgcgatgttt cgcttggtgg
     3781 tcgaatgggc aggtagccgg atcaagcgta tgcagccgcc gcattgcatc agccatgatg
     3841 gatactttct cggcaggagc aaggtgagat gacaggagat cctgccccgg cacttcgccc
     3901 aatagcagcc agtcccttcc cgcttcagtg acaacgtcga gcacagctgc gcaaggaacg
     3961 cccgtcgtgg ccagccacga tagccgcgct gcctcgtctt gcagttcatt cagggcaccg
     4021 gacaggtcgg tcttgacaaa aagaaccggg cgcccctgcg ctgacagccg gaacacggcg
     4081 gcatcagagc agccgattgt ctgttgtgcc cagtcatagc cgaatagcct ctccacccaa
     4141 gcggccggag aacctgcgtg caatccatct tgttcaatca tgcgaaacga tcctcatcct
     4201 gtctcttgat cgatctttgc aaaagcctag gcctccaaaa aagcctcctc actacttctg
     4261 gaatagctca gaggccgagg cggcctcggc ctctgcataa ataaaaaaaa ttagtcagcc
     4321 atggggcgga gaatgggcgg aactgggcgg agttaggggc gggatgggcg gagttagggg
     4381 cgggactatg gttgctgact aattgagatg catgctttgc atacttctgc ctgctgggga
     4441 gcctggggac tttccacacc tggttgctga ctaattgaga tgcatgcttt gcatacttct
     4501 gcctgctggg gagcctgggg actttccaca ccctaactga cacacattcc acagctggtt
     4561 ctttccgcct caggactctt cctttttcaa taaatcaatc taaagtatat atgagtaaac
     4621 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     4681 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     4741 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     4801 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     4861 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     4921 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     4981 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     5041 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     5101 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     5161 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     5221 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     5281 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     5341 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     5401 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     5461 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     5521 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     5581 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgcgc cctgtagcgg
     5641 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     5701 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     5761 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     5821 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     5881 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     5941 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
     6001 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa
     6061 aatattaacg cttacaattt ac
//
