LOCUS       sequence-315982-        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"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2268..2297
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2313..2330
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2353..2370)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2359..2583
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2629..3057
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2716..2735)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2926..2947
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3066..3086)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3071..3400
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3251..3386
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3313..3332
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3467..4261
                     /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(3521..3540)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4131..4150
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4437..4558
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4474..4493)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4528..4547
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4607..4623)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4607..4623)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4620..4642)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4631..4647
                     /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(4655..4661,4662..4679,4680..4685))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4700..4721
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4838..4855)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5009..5597)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5089..5108)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5768..6559,6560..6628))
                     /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     6391..6410
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6629..6733)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagt
      901 taagcttggt accgagctcg gatccactag tccagtgtgg tggaattgcc accatgtcct
      961 ttattccagt ggccgaggac tccgactttc ccatccaaaa cctgccctat ggtgttttct
     1021 ccactcaaag caacccaaag ccacggattg gtgtagccat cggtgaccag atcttggacc
     1081 tgagtgtcat taaacacctc tttaccggac ctgccctttc caaacatcaa catgtcttcg
     1141 atgagacaac tctcagtaac ttcatgggtc tgggtcaagc tgcatggaag gaggcaagag
     1201 catccttaca gaacttactg tctgccagcc aagcccggct cagagatgac aaggagcttc
     1261 ggcagcgtgc attcacctcc caggcttctg cgacaatgca ccttcctgct accataggag
     1321 actacacgga cttctactct tctcggcagc atgccaccaa tgttggcatt atgttcagag
     1381 gcaaggagaa tgcgctgttg ccaaattggc tccacttacc tgtgggatac catggccgag
     1441 cttcctccat tgtggtatct ggaaccccga ttcgaagacc catggggcag atgagacctg
     1501 ataactcaaa gcctcctgtg tatggtgcct gcagactctt agacatggag ttggaaatgg
     1561 ctttcttcgt aggccctggg aacagattcg gagagccaat ccccatttcc aaagcccatg
     1621 aacacatttt cgggatggtc ctcatgaacg actggagcgc acgagacatc cagcaatggg
     1681 agtacgtccc acttgggcca ttcctgggga aaagctttgg aaccacaatc tccccgtggg
     1741 tggtgcctat ggatgccctc atgccctttg tggtgccaaa cccaaagcag gaccccaagc
     1801 ccttgccata tctctgccac agccagccct acacatttga tatcaacctg tctgtctctt
     1861 tgaaaggaga aggaatgagc caggcggcta ccatctgcag gtctaacttt aagcacatgt
     1921 actggaccat gctgcagcaa ctcacacacc actctgttaa tggatgcaac ctgagacctg
     1981 gggacctctt ggcttctgga accatcagtg gatcagaccc tgaaagcttt ggctccatgc
     2041 tggaactgtc ctggaaggga acaaaggcca tcgatgtgga gcaggggcag accaggacct
     2101 tcctgctgga cggcgatgaa gtcatcataa caggtcactg ccagggggac ggctaccgtg
     2161 ttggctttgg ccagtgtgct gggaaagtgc tgcctgccct ttcaccagcc aattctgcag
     2221 atatccagca cagtggcggc cgctcgagtc tagagggccc gcggttcgaa caaaaactca
     2281 tctcagaaga ggatctgaat atgcataccg gtcatcatca ccatcaccat tgagtttaaa
     2341 cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc
     2401 ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg
     2461 aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg
     2521 acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta
     2581 tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta
     2641 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     2701 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     2761 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     2821 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
     2881 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     2941 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
     3001 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat
     3061 tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag
     3121 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
     3181 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
     3241 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     3301 actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa
     3361 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat
     3421 atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga
     3481 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc
     3541 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc
     3601 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc
     3661 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac
     3721 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc
     3781 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac
     3841 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg
     3901 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct
     3961 cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt
     4021 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg
     4081 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac
     4141 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg
     4201 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg
     4261 agcgggactc tggggttcgc gaaatgaccg accaagcgac gcccaacctg ccatcacgag
     4321 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
     4381 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact
     4441 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     4501 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     4561 atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc
     4621 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     4681 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     4741 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     4801 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     4861 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     4921 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     4981 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     5041 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     5101 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     5161 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     5221 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     5281 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     5341 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     5401 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     5461 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     5521 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     5581 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     5641 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     5701 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     5761 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     5821 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     5881 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     5941 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     6001 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     6061 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     6121 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     6181 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     6241 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     6301 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     6361 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     6421 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     6481 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     6541 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     6601 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     6661 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     6721 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtc
//
