LOCUS       sequence-447940-        5209 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express HNF4A in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5209)
  AUTHORS   Nakagawa S, Yamaguchi K, Takane K, Tabata S, Ikenoue T, Furukawa Y
  TITLE     Wnt/beta-catenin signaling regulates amino acid metabolism through 
            the suppression of CEBPA and FOXA1 in liver cancer cells.
  JOURNAL   Commun Biol. 2024 Apr 29;7(1):510. doi: 10.1038/s42003-024-06202-9.
  PUBMED    38684876
REFERENCE   2  (bases 1 to 5209)
  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"; journalName: "Commun 
            Biol."; date: "2024-04-29"; volume: "7(1):510. doi"; pages: " 
            10.1038/s42003-024-06202-9"
COMMENT     Sequence Label: pCMV-Myc-HNF4A
FEATURES             Location/Qualifiers
     source          1..5209
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        27..330
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        331..534
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     485..505
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     531..555
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          672..768
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     polyA_signal    2348..2482
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2372..2391)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2426..2445
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2593..2609)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2593..2609)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2606..2628)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2617..2633
                     /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(2641..2647,2648..2665,2666..2671))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2686..2707
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2824..2841)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2995..3583)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3075..3094)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3754..4545,4546..4614))
                     /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     4377..4396
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4615..4719)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4787..4805
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4843..4865)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4965..4984
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5178..5200
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5193..5209
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gagttcgagc ttgcatgcct gcaggtcgtt acataactta cggtaaatgg cccgcctggc
       61 tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg
      121 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg
      181 gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa
      241 tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac
      301 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg
      361 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg
      421 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca
      481 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta
      541 gtgaaccgtc agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagacac
      601 cgggaccgat ccagcctccg gactctagag gatccggtac tagaggaact gaaaaaccag
      661 aaagttaact ggtaagttta gtctttttgt cttttatttc aggtcccgga tccggtggtg
      721 gtgcaaatca aagaactgct cctcagtgga tgttgccttt acttctaggc ctgtacggaa
      781 gtgttacttc tgctctaaaa gctgcggaat tgtacccgcg ggcccaccat ggcatcaatg
      841 cagaagctga tctcagagga ggacctgctt atggccatgg aggcccgaat tcggatgcga
      901 ctctccaaaa ccctcgtcga catggacatg gccgactaca gtgctgcact ggacccagcc
      961 tacaccaccc tggaatttga gaatgtgcag gtgttgacga tgggcaacga cacgtcccca
     1021 tcagaaggca ccaacctcaa cgcgcccaac agcctgggtg tcagcgccct gtgtgccatc
     1081 tgcggggacc gggccacggg caaacactac ggtgcctcga gctgtgacgg ctgcaagggc
     1141 ttcttccgga ggagcgtgcg gaagaaccac atgtactcct gcagatttag ccggcagtgc
     1201 gtggtggaca aagacaagag gaaccagtgc cgctactgca ggctcaagaa atgcttccgg
     1261 gctggcatga agaaggaagc cgtccagaat gagcgggacc ggatcagcac tcgaaggtca
     1321 agctatgagg acagcagcct gccctccatc aatgcgctcc tgcaggcgga ggtcctgtcc
     1381 cgacagatca cctcccccgt ctccgggatc aacggcgaca ttcgggcgaa gaagattgcc
     1441 agcatcgcag atgtgtgtga gtccatgaag gagcagctgc tggttctcgt tgagtgggcc
     1501 aagtacatcc cagctttctg cgagctcccc ctggacgacc aggtggccct gctcagagcc
     1561 catgctggcg agcacctgct gctcggagcc accaagagat ccatggtgtt caaggacgtg
     1621 ctgctcctag gcaatgacta cattgtccct cggcactgcc cggagctggc ggagatgagc
     1681 cgggtgtcca tacgcatcct tgacgagctg gtgctgccct tccaggagct gcagatcgat
     1741 gacaatgagt atgcctacct caaagccatc atcttctttg acccagatgc caaggggctg
     1801 agcgatccag ggaagatcaa gcggctgcgt tcccaggtgc aggtgagctt ggaggactac
     1861 atcaacgacc gccagtatga ctcgcgtggc cgctttggag agctgctgct gctgctgccc
     1921 accttgcaga gcatcacctg gcagatgatc gagcagatcc agttcatcaa gctcttcggc
     1981 atggccaaga ttgacaacct gttgcaggag atgctgctgg gagggtcccc cagcgatgca
     2041 ccccatgccc accaccccct gcaccctcac ctgatgcagg aacatatggg aaccaacgtc
     2101 atcgttgcca acacaatgcc cactcacctc agcaacggac agatgtgtga gtggccccga
     2161 cccaggggac aggcagccac ccctgagacc ccacagccct caccgccagg tggctcaggg
     2221 tctgagccct ataagctcct gccgggagcc gtcgccacaa tcgtcaagcc cctctctgcc
     2281 atcccccagc cgaccatcac caagcaggaa gttatctaga gatctctcga ggtaccgcgg
     2341 ccgcggggat ccagacatga taagatacat tgatgagttt ggacaaacca caactagaat
     2401 gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat
     2461 tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca
     2521 gggggaggtg tgggaggttt tttcggatcc tctagagtcg atctgcaggc atgctagctt
     2581 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     2641 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     2701 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     2761 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     2821 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     2881 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     2941 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     3001 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     3061 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     3121 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     3181 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     3241 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     3301 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     3361 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     3421 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     3481 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     3541 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     3601 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     3661 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     3721 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     3781 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     3841 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     3901 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     3961 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     4021 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     4081 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     4141 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     4201 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     4261 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     4321 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     4381 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     4441 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     4501 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     4561 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     4621 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     4681 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     4741 acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac
     4801 gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct
     4861 cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg
     4921 cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat
     4981 tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata
     5041 ccgcatcagg cgccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg
     5101 ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg
     5161 ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagt
//
