LOCUS       sequence-320638-        6764 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expressing mouse wild type FAH in mammalian cells .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6764)
  AUTHORS   Yang S, Siepka SM, Cox KH, Kumar V, de Groot M, Chelliah Y, Chen J, 
            Tu B, Takahashi JS
  TITLE     Tissue-specific FAH deficiency alters sleep-wake patterns and 
            results in chronic tyrosinemia in mice.
  JOURNAL   Proc Natl Acad Sci U S A. 2019 Oct 29;116(44):22229-22236. doi: 
            10.1073/pnas.1904485116. Epub 2019 Oct 14.
  PUBMED    31611405
REFERENCE   2  (bases 1 to 6764)
  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: 
            "10.1073/pnas.1904485116"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2019-10-29- 29"; volume: "116"; issue: "44"; pages: 
            "22229-22236"
COMMENT     Sequence Label: pcDNA-Fah_W
FEATURES             Location/Qualifiers
     source          1..6764
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        46..425
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        426..629
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     580..600
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     674..693
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        674..692
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2079..2108
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2124..2141
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2164..2181)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2170..2394
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2440..2868
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2527..2546)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2737..2758
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2877..2897)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2882..3211
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3062..3197
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3124..3143
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3278..4072
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3332..3351)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3942..3961
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4248..4369
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4285..4304)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4339..4358
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4418..4434)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4418..4434)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4431..4453)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4442..4458
                     /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(4466..4472,4473..4490,4491..4496))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4511..4532
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4649..4666)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4820..5408)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4900..4919)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5579..6370,6371..6439))
                     /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     6202..6221
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6440..6544)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6619..6638)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 ggcgttttgc gctgcttcgc gatgtacggg ccagatatac gcgttgacat tgattattga
       61 ctagttatta atagtaatca attacggggt cattagttca tagcccatat atggagttcc
      121 gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat
      181 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc
      241 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
      301 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt
      361 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
      421 ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg
      481 gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac
      541 gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg
      601 tacggtggga ggtctatata agcagagctc tctggctaac tagagaaccc actgcttact
      661 ggcttatcga aattaatacg actcactata gggagaccca agctggctag ttaagcttgg
      721 taccgagctc ggatccacta gtccagtgtg gtggaattgc caccatgtcc tttattccag
      781 tggccgagga ctccgacttt cccatccaaa acctgcccta tggtgttttc tccactcaaa
      841 gcaacccaaa gccacggatt ggtgtagcca tcggtgacca gatcttggac ctgagtgtca
      901 ttaaacacct ctttaccgga cctgcccttt ccaaacatca acatgtcttc gatgagacaa
      961 ctctcaataa cttcatgggt ctgggtcaag ctgcatggaa ggaggcaaga gcatccttac
     1021 agaacttact gtctgccagc caagcccggc tcagagatga caaggagctt cggcagcgtg
     1081 cattcacctc ccaggcttct gcgacaatgc accttcctgc taccatagga gactacacgg
     1141 acttctactc ttctcggcag catgccacca atgttggcat tatgttcaga ggcaaggaga
     1201 atgcgctgtt gccaaattgg ctccacttac ctgtgggata ccatggccga gcttcctcca
     1261 ttgtggtatc tggaaccccg attcgaagac ccatggggca gatgagacct gataactcaa
     1321 agcctcctgt gtatggtgcc tgcagactct tagacatgga gttggaaatg gctttcttcg
     1381 taggccctgg gaacagattc ggagagccaa tccccatttc caaagcccat gaacacattt
     1441 tcgggatggt cctcatgaac gactggagcg cacgagacat ccagcaatgg gagtacgtcc
     1501 cacttgggcc attcctgggg aaaagctttg gaaccacaat ctccccgtgg gtggtgccta
     1561 tggatgccct catgcccttt gtggtgccaa acccaaagca ggaccccaag cccttgccat
     1621 atctctgcca cagccagccc tacacatttg atatcaacct gtctgtctct ttgaaaggag
     1681 aaggaatgag ccaggcggct accatctgca ggtctaactt taagcacatg tactggacca
     1741 tgctgcagca actcacacac cactctgtta atggatgcaa cctgagacct ggggacctct
     1801 tggcttctgg aaccatcagt ggatcagacc ctgaaagctt tggctccatg ctggaactgt
     1861 cctggaaggg aacaaaggcc atcgatgtgg agcaggggca gaccaggacc ttcctgctgg
     1921 acggcgatga agtcatcata acaggtcact gccaggggga cggctaccgt gttggctttg
     1981 gccagtgtgc tgggaaagtg ctgcctgccc tttcaccagc caattctgca gatatccagc
     2041 acagtggcgg ccgctcgagt ctagagggcc cgcggttcga acaaaaactc atctcagaag
     2101 aggatctgaa tatgcatacc ggtcatcatc accatcacca ttgagtttaa acccgctgat
     2161 cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt
     2221 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat
     2281 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg
     2341 gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg
     2401 aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat
     2461 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     2521 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     2581 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     2641 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     2701 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     2761 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     2821 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa
     2881 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     2941 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     3001 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     3061 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     3121 ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg
     3181 aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt
     3241 cggatctgat caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca
     3301 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac
     3361 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt
     3421 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc
     3481 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg
     3541 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc
     3601 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc
     3661 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat
     3721 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc
     3781 cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca
     3841 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga
     3901 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat
     3961 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc
     4021 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact
     4081 ctggggttcg cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt
     4141 ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga
     4201 tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg
     4261 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     4321 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta
     4381 taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     4441 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     4501 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     4561 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     4621 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     4681 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     4741 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     4801 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     4861 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     4921 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     4981 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     5041 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     5101 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     5161 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     5221 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     5281 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     5341 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     5401 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     5461 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     5521 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     5581 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     5641 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     5701 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     5761 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     5821 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     5881 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     5941 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     6001 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     6061 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     6121 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     6181 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     6241 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     6301 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     6361 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     6421 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     6481 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     6541 cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat ctcccgatcc
     6601 cctatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agtatctgct
     6661 ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag
     6721 gcaaggcttg accgacaatt gcatgaagaa tctgcttagg gtta
//
