LOCUS       sequence-320634-        6764 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Swingshift mutant 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_S
FEATURES             Location/Qualifiers
     source          1..6764
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        122..501
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        502..705
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     656..676
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     750..769
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        750..768
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2155..2184
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2200..2217
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2240..2257)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2246..2470
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2516..2944
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2603..2622)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2813..2834
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2953..2973)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2958..3287
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3138..3273
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3200..3219
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3354..4148
                     /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(3408..3427)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4018..4037
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4324..4445
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4361..4380)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4415..4434
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4494..4510)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4494..4510)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4507..4529)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4518..4534
                     /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(4542..4548,4549..4566,4567..4572))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4587..4608
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4725..4742)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4896..5484)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4976..4995)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5655..6446,6447..6515))
                     /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     6278..6297
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6516..6620)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6695..6714)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 tagtgcgcga gcaaaattta agctacaaca aggcaaggct tgaccgacaa ttgcatgaag
       61 aatctgctta gggttaggcg ttttgcgctg cttcgcgatg tacgggccag atatacgcgt
      121 tgacattgat tattgactag ttattaatag taatcaatta cggggtcatt agttcatagc
      181 ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc
      241 aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg
      301 actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat
      361 caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc
      421 tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta catctacgta
      481 ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag
      541 cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt
      601 tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa
      661 atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctctctg gctaactaga
      721 gaacccactg cttactggct tatcgaaatt aatacgactc actataggga gacccaagct
      781 ggctagttaa gcttggtacc gagctcggat ccactagtcc agtgtggtgg aattgccacc
      841 atgtccttta ttccagtggc cgaggactcc gactttccca tccaaaacct gccctatggt
      901 gttttctcca ctcaaagcaa cccaaagcca cggattggtg tagccatcgg tgaccagatc
      961 ttggacctga gtgtcattaa acacctcttt accggacctg ccctttccaa acatcaacat
     1021 gtcttcgatg agacaactct cagtaacttc atgggtctgg gtcaagctgc atggaaggag
     1081 gcaagagcat ccttacagaa cttactgtct gccagccaag cccggctcag agatgacaag
     1141 gagcttcggc agcgtgcatt cacctcccag gcttctgcga caatgcacct tcctgctacc
     1201 ataggagact acacggactt ctactcttct cggcagcatg ccaccaatgt tggcattatg
     1261 ttcagaggca aggagaatgc gctgttgcca aattggctcc acttacctgt gggataccat
     1321 ggccgagctt cctccattgt ggtatctgga accccgattc gaagacccat ggggcagatg
     1381 agacctgata actcaaagcc tcctgtgtat ggtgcctgca gactcttaga catggagttg
     1441 gaaatggctt tcttcgtagg ccctgggaac agattcggag agccaatccc catttccaaa
     1501 gcccatgaac acattttcgg gatggtcctc atgaacgact ggagcgcacg agacatccag
     1561 caatgggagt acgtcccact tgggccattc ctggggaaaa gctttggaac cacaatctcc
     1621 ccgtgggtgg tgcctatgga tgccctcatg ccctttgtgg tgccaaaccc aaagcaggac
     1681 cccaagccct tgccatatct ctgccacagc cagccctaca catttgatat caacctgtct
     1741 gtctctttga aaggagaagg aatgagccag gcggctacca tctgcaggtc taactttaag
     1801 cacatgtact ggaccatgct gcagcaactc acacaccact ctgttaatgg atgcaacctg
     1861 agacctgggg acctcttggc ttctggaacc atcagtggat cagaccctga aagctttggc
     1921 tccatgctgg aactgtcctg gaagggaaca aaggccatcg atgtggagca ggggcagacc
     1981 aggaccttcc tgctggacgg cgatgaagtc atcataacag gtcactgcca gggggacggc
     2041 taccgtgttg gctttggcca gtgtgctggg aaagtgctgc ctgccctttc accagccaat
     2101 tctgcagata tccagcacag tggcggccgc tcgagtctag agggcccgcg gttcgaacaa
     2161 aaactcatct cagaagagga tctgaatatg cataccggtc atcatcacca tcaccattga
     2221 gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc
     2281 ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa
     2341 aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg
     2401 gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg
     2461 ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta tccccacgcg
     2521 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     2581 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     2641 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     2701 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     2761 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     2821 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     2881 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     2941 aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     3001 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     3061 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     3121 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     3181 atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat
     3241 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag
     3301 cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt cgcatgattg
     3361 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg
     3421 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg
     3481 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg
     3541 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg
     3601 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc
     3661 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc
     3721 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc
     3781 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc
     3841 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg
     3901 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct
     3961 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt
     4021 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc
     4081 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt
     4141 tcttctgagc gggactctgg ggttcgcgaa atgaccgacc aagcgacgcc caacctgcca
     4201 tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc
     4261 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac
     4321 cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     4381 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     4441 tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag
     4501 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     4561 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     4621 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     4681 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     4741 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     4801 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     4861 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     4921 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     4981 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     5041 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     5101 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     5161 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     5221 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     5281 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     5341 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     5401 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     5461 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     5521 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     5581 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     5641 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     5701 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     5761 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     5821 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     5881 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     5941 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     6001 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     6061 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     6121 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     6181 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     6241 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     6301 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     6361 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     6421 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     6481 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     6541 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     6601 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacggatcg
     6661 ggagatctcc cgatccccta tggtgcactc tcagtacaat ctgctctgat gccgcatagt
     6721 taagccagta tctgctccct gcttgtgtgt tggaggtcgc tgag
//
