LOCUS       sequence-320643-        6302 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expressing mouse wildtype FAH in bacterial cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6302)
  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 6302)
  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: pGST-Fah_W
FEATURES             Location/Qualifiers
     source          1..6302
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(222..227,228..243,244..250)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    258..274
                     /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)."
     CDS             297..950
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     908..930
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             963..983
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(2391..2413)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2558..2576)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2644..2748
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2749..2817,2818..3609)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2967..2986)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3780..4368
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4269..4288
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4522..4539
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        4612..4689
                     /gene="lacI"
                     /label=lacI promoter
     CDS             4690..5772
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(4709..4728)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(5821..5826,5827..5844,5845..5851)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5859..5875
                     /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)."
     primer_bind     5883..5899
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5883..5899
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             join(5895..6302,1..120)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(5915..5932)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5915..5931)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5924..5946)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 gaaggccaga cgcgaattat ttttgatggc gttggaatta gcttatcgac tgcacggtgc
       61 accaatgctt ctggcgtcag gcagccatcg gaagctgtgg tatggctgtg caggtcgtaa
      121 atcactgcat aattcgtgtc gctcaaggcg cactcccgtt ctggataatg ttttttgcgc
      181 cgacatcata acggttctgg caaatattct gaaatgagct gttgacaatt aatcatcggc
      241 tcgtataatg tgtggaattg tgagcggata acaatttcac acaggaaaca gtattcatgt
      301 cccctatact aggttattgg aaaattaagg gccttgtgca acccactcga cttcttttgg
      361 aatatcttga agaaaaatat gaagagcatt tgtatgagcg cgatgaaggt gataaatggc
      421 gaaacaaaaa gtttgaattg ggtttggagt ttcccaatct tccttattat attgatggtg
      481 atgttaaatt aacacagtct atggccatca tacgttatat agctgacaag cacaacatgt
      541 tgggtggttg tccaaaagag cgtgcagaga tttcaatgct tgaaggagcg gttttggata
      601 ttagatacgg tgtttcgaga attgcatata gtaaagactt tgaaactctc aaagttgatt
      661 ttcttagcaa gctacctgaa atgctgaaaa tgttcgaaga tcgtttatgt cataaaacat
      721 atttaaatgg tgatcatgta acccatcctg acttcatgtt gtatgacgct cttgatgttg
      781 ttttatacat ggacccaatg tgcctggatg cgttcccaaa attagtttgt tttaaaaaac
      841 gtattgaagc tatcccacaa attgataagt acttgaaatc cagcaagtat atagcatggc
      901 ctttgcaggg ctggcaagcc acgtttggtg gtggcgacca tcctccaaaa atcgatacga
      961 ccgaaaacct gtattttcag ggcgccatgg atccggaatt catgtccttt attccagtgg
     1021 ccgaggactc cgactttccc atccaaaacc tgccctatgg tgttttctcc actcaaagca
     1081 acccaaagcc acggattggt gtagccatcg gtgaccagat cttggacctg agtgtcatta
     1141 aacacctctt taccggacct gccctttcca aacatcaaca tgtcttcgat gagacaactc
     1201 tcaataactt catgggtctg ggtcaagctg catggaagga ggcaagagca tccttacaga
     1261 acttactgtc tgccagccaa gcccggctca gagatgacaa ggagcttcgg cagcgtgcat
     1321 tcacctccca ggcttctgcg acaatgcacc ttcctgctac cataggagac tacacggact
     1381 tctactcttc tcggcagcat gccaccaatg ttggcattat gttcagaggc aaggagaatg
     1441 cgctgttgcc aaattggctc cacttacctg tgggatacca tggccgagct tcctccattg
     1501 tggtatctgg aaccccgatt cgaagaccca tggggcagat gagacctgat aactcaaagc
     1561 ctcctgtgta tggtgcctgc agactcttag acatggagtt ggaaatggct ttcttcgtag
     1621 gccctgggaa cagattcgga gagccaatcc ccatttccaa agcccatgaa cacattttcg
     1681 ggatggtcct catgaacgac tggagcgcac gagacatcca gcaatgggag tacgtcccac
     1741 ttgggccatt cctggggaaa agctttggaa ccacaatctc cccgtgggtg gtgcctatgg
     1801 atgccctcat gccctttgtg gtgccaaacc caaagcagga ccccaagccc ttgccatatc
     1861 tctgccacag ccagccctac acatttgata tcaacctgtc tgtctctttg aaaggagaag
     1921 gaatgagcca ggcggctacc atctgcaggt ctaactttaa gcacatgtac tggaccatgc
     1981 tgcagcaact cacacaccac tctgttaatg gatgcaacct gagacctggg gacctcttgg
     2041 cttctggaac catcagtgga tcagaccctg aaagctttgg ctccatgctg gaactgtcct
     2101 ggaagggaac aaaggccatc gatgtggagc aggggcagac caggaccttc ctgctggacg
     2161 gcgatgaagt catcataaca ggtcactgcc agggggacgg ctaccgtgtt ggctttggcc
     2221 agtgtgctgg gaaagtgctg cctgcccttt caccagcctg aaattcaaag gcctacgtcg
     2281 acgagctcaa ctagtgcggc cgctttcgaa tctagagcct gcagtctcga gcggccgcat
     2341 cgtgactgac tgacgatctg cctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac
     2401 atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc
     2461 cgtcagggcg cgtcagcggg tgttggcggg tgtcggggcg cagccatgac ccagtcacgt
     2521 agcgatagcg gagtgtataa ttcttgaaga cgaaagggcc tcgtgatacg cctattttta
     2581 taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat
     2641 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     2701 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     2761 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     2821 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     2881 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     2941 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tgttgacgcc
     3001 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     3061 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     3121 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     3181 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     3241 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgcagcaatg
     3301 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     3361 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     3421 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     3481 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     3541 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     3601 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     3661 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     3721 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     3781 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     3841 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     3901 agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc
     3961 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     4021 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     4081 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     4141 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     4201 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     4261 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     4321 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     4381 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     4441 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     4501 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg
     4561 cggtattttc tccttacgca tctgtgcggt atttcacacc gcataaattc cgacaccatc
     4621 gaatggcgca aaacctttcg cggtatggca tgatagcgcc cggaagagag tcaattcagg
     4681 gtggtgaatg tgaaaccagt aacgttatac gatgtcgcag agtatgccgg tgtctcttat
     4741 cagaccgttt cccgcgtggt gaaccaggcc agccacgttt ctgcgaaaac gcgggaaaaa
     4801 gtggaagcgg cgatggcgga gctgaattac attcccaacc gcgtggcaca acaactggcg
     4861 ggcaaacagt cgttgctgat tggcgttgcc acctccagtc tggccctgca cgcgccgtcg
     4921 caaattgtcg cggcgattaa atctcgcgcc gatcaactgg gtgccagcgt ggtggtgtcg
     4981 atggtagaac gaagcggcgt cgaagcctgt aaagcggcgg tgcacaatct tctcgcgcaa
     5041 cgcgtcagtg ggctgatcat taactatccg ctggatgacc aggatgccat tgctgtggaa
     5101 gctgcctgca ctaatgttcc ggcgttattt cttgatgtct ctgaccagac acccatcaac
     5161 agtattattt tctcccatga agacggtacg cgactgggcg tggagcatct ggtcgcattg
     5221 ggtcaccagc aaatcgcgct gttagcgggc ccattaagtt ctgtctcggc gcgtctgcgt
     5281 ctggctggct ggcataaata tctcactcgc aatcaaattc agccgatagc ggaacgggaa
     5341 ggcgactgga gtgccatgtc cggttttcaa caaaccatgc aaatgctgaa tgagggcatc
     5401 gttcccactg cgatgctggt tgccaacgat cagatggcgc tgggcgcaat gcgcgccatt
     5461 accgagtccg ggctgcgcgt tggtgcggat atctcggtag tgggatacga cgataccgaa
     5521 gacagctcat gttatatccc gccgttaacc accatcaaac aggattttcg cctgctgggg
     5581 caaaccagcg tggaccgctt gctgcaactc tctcagggcc aggcggtgaa gggcaatcag
     5641 ctgttgcccg tctcactggt gaaaagaaaa accaccctgg cgcccaatac gcaaaccgcc
     5701 tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa
     5761 agcgggcagt gagcgcaacg caattaatgt aagttagctc actcattagg caccccaggc
     5821 tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca
     5881 cacaggaaac agctatgacc atgattacgg attcactggc cgtcgtttta caacgtcgtg
     5941 actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca
     6001 gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga
     6061 atggcgaatg gcgctttgcc tggtttccgg caccagaagc ggtgccggaa agctggctgg
     6121 agtgcgatct tcctgaggcc gatactgtcg tcgtcccctc aaactggcag atgcacggtt
     6181 acgatgcgcc catctacacc aacgtgacct atcccattac ggtcaatccg ccgtttgttc
     6241 ccacggagaa tccgacgggt tgttactcgc tcacatttaa tgttgatgaa agctggctac
     6301 ag
//
