LOCUS       sequence-320639-        6302 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expressing mouse swingshift mutant 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_S
FEATURES             Location/Qualifiers
     source          1..6302
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(124..129,130..147,148..154)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    162..178
                     /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     186..202
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     186..202
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             198..725
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(218..235)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(218..234)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(227..249)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(827..832,833..848,849..855)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    863..879
                     /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             902..1555
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     1513..1535
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             1568..1588
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(2996..3018)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3163..3181)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3249..3353
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3354..3422,3423..4214)
                     /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(3572..3591)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4385..4973
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4874..4893
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5127..5144
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        5217..5294
                     /gene="lacI"
                     /label=lacI promoter
     CDS             join(5295..6302,1..75)
                     /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(5314..5333)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg
       61 gaaagcgggc agtgagcgca acgcaattaa tgtaagttag ctcactcatt aggcacccca
      121 ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt
      181 tcacacagga aacagctatg accatgatta cggattcact ggccgtcgtt ttacaacgtc
      241 gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg
      301 ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc
      361 tgaatggcga atggcgcttt gcctggtttc cggcaccaga agcggtgccg gaaagctggc
      421 tggagtgcga tcttcctgag gccgatactg tcgtcgtccc ctcaaactgg cagatgcacg
      481 gttacgatgc gcccatctac accaacgtga cctatcccat tacggtcaat ccgccgtttg
      541 ttcccacgga gaatccgacg ggttgttact cgctcacatt taatgttgat gaaagctggc
      601 tacaggaagg ccagacgcga attatttttg atggcgttgg aattagctta tcgactgcac
      661 ggtgcaccaa tgcttctggc gtcaggcagc catcggaagc tgtggtatgg ctgtgcaggt
      721 cgtaaatcac tgcataattc gtgtcgctca aggcgcactc ccgttctgga taatgttttt
      781 tgcgccgaca tcataacggt tctggcaaat attctgaaat gagctgttga caattaatca
      841 tcggctcgta taatgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagtatt
      901 catgtcccct atactaggtt attggaaaat taagggcctt gtgcaaccca ctcgacttct
      961 tttggaatat cttgaagaaa aatatgaaga gcatttgtat gagcgcgatg aaggtgataa
     1021 atggcgaaac aaaaagtttg aattgggttt ggagtttccc aatcttcctt attatattga
     1081 tggtgatgtt aaattaacac agtctatggc catcatacgt tatatagctg acaagcacaa
     1141 catgttgggt ggttgtccaa aagagcgtgc agagatttca atgcttgaag gagcggtttt
     1201 ggatattaga tacggtgttt cgagaattgc atatagtaaa gactttgaaa ctctcaaagt
     1261 tgattttctt agcaagctac ctgaaatgct gaaaatgttc gaagatcgtt tatgtcataa
     1321 aacatattta aatggtgatc atgtaaccca tcctgacttc atgttgtatg acgctcttga
     1381 tgttgtttta tacatggacc caatgtgcct ggatgcgttc ccaaaattag tttgttttaa
     1441 aaaacgtatt gaagctatcc cacaaattga taagtacttg aaatccagca agtatatagc
     1501 atggcctttg cagggctggc aagccacgtt tggtggtggc gaccatcctc caaaaatcga
     1561 tacgaccgaa aacctgtatt ttcagggcgc catggatccg gaattcatgt cctttattcc
     1621 agtggccgag gactccgact ttcccatcca aaacctgccc tatggtgttt tctccactca
     1681 aagcaaccca aagccacgga ttggtgtagc catcggtgac cagatcttgg acctgagtgt
     1741 cattaaacac ctctttaccg gacctgccct ttccaaacat caacatgtct tcgatgagac
     1801 aactctcagt aacttcatgg gtctgggtca agctgcatgg aaggaggcaa gagcatcctt
     1861 acagaactta ctgtctgcca gccaagcccg gctcagagat gacaaggagc ttcggcagcg
     1921 tgcattcacc tcccaggctt ctgcgacaat gcaccttcct gctaccatag gagactacac
     1981 ggacttctac tcttctcggc agcatgccac caatgttggc attatgttca gaggcaagga
     2041 gaatgcgctg ttgccaaatt ggctccactt acctgtggga taccatggcc gagcttcctc
     2101 cattgtggta tctggaaccc cgattcgaag acccatgggg cagatgagac ctgataactc
     2161 aaagcctcct gtgtatggtg cctgcagact cttagacatg gagttggaaa tggctttctt
     2221 cgtaggccct gggaacagat tcggagagcc aatccccatt tccaaagccc atgaacacat
     2281 tttcgggatg gtcctcatga acgactggag cgcacgagac atccagcaat gggagtacgt
     2341 cccacttggg ccattcctgg ggaaaagctt tggaaccaca atctccccgt gggtggtgcc
     2401 tatggatgcc ctcatgccct ttgtggtgcc aaacccaaag caggacccca agcccttgcc
     2461 atatctctgc cacagccagc cctacacatt tgatatcaac ctgtctgtct ctttgaaagg
     2521 agaaggaatg agccaggcgg ctaccatctg caggtctaac tttaagcaca tgtactggac
     2581 catgctgcag caactcacac accactctgt taatggatgc aacctgagac ctggggacct
     2641 cttggcttct ggaaccatca gtggatcaga ccctgaaagc tttggctcca tgctggaact
     2701 gtcctggaag ggaacaaagg ccatcgatgt ggagcagggg cagaccagga ccttcctgct
     2761 ggacggcgat gaagtcatca taacaggtca ctgccagggg gacggctacc gtgttggctt
     2821 tggccagtgt gctgggaaag tgctgcctgc cctttcacca gcctgaaatt caaaggccta
     2881 cgtcgacgag ctcaactagt gcggccgctt tcgaatctag agcctgcagt ctcgagcggc
     2941 cgcatcgtga ctgactgacg atctgcctcg cgcgtttcgg tgatgacggt gaaaacctct
     3001 gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac
     3061 aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggcgcagcc atgacccagt
     3121 cacgtagcga tagcggagtg tataattctt gaagacgaaa gggcctcgtg atacgcctat
     3181 ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg
     3241 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     3301 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     3361 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     3421 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     3481 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     3541 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtgttg
     3601 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     3661 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     3721 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     3781 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     3841 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgcag
     3901 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     3961 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     4021 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     4081 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     4141 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     4201 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     4261 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     4321 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     4381 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     4441 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     4501 gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc
     4561 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     4621 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     4681 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     4741 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     4801 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     4861 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     4921 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     4981 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     5041 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     5101 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
     5161 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata aattccgaca
     5221 ccatcgaatg gcgcaaaacc tttcgcggta tggcatgata gcgcccggaa gagagtcaat
     5281 tcagggtggt gaatgtgaaa ccagtaacgt tatacgatgt cgcagagtat gccggtgtct
     5341 cttatcagac cgtttcccgc gtggtgaacc aggccagcca cgtttctgcg aaaacgcggg
     5401 aaaaagtgga agcggcgatg gcggagctga attacattcc caaccgcgtg gcacaacaac
     5461 tggcgggcaa acagtcgttg ctgattggcg ttgccacctc cagtctggcc ctgcacgcgc
     5521 cgtcgcaaat tgtcgcggcg attaaatctc gcgccgatca actgggtgcc agcgtggtgg
     5581 tgtcgatggt agaacgaagc ggcgtcgaag cctgtaaagc ggcggtgcac aatcttctcg
     5641 cgcaacgcgt cagtgggctg atcattaact atccgctgga tgaccaggat gccattgctg
     5701 tggaagctgc ctgcactaat gttccggcgt tatttcttga tgtctctgac cagacaccca
     5761 tcaacagtat tattttctcc catgaagacg gtacgcgact gggcgtggag catctggtcg
     5821 cattgggtca ccagcaaatc gcgctgttag cgggcccatt aagttctgtc tcggcgcgtc
     5881 tgcgtctggc tggctggcat aaatatctca ctcgcaatca aattcagccg atagcggaac
     5941 gggaaggcga ctggagtgcc atgtccggtt ttcaacaaac catgcaaatg ctgaatgagg
     6001 gcatcgttcc cactgcgatg ctggttgcca acgatcagat ggcgctgggc gcaatgcgcg
     6061 ccattaccga gtccgggctg cgcgttggtg cggatatctc ggtagtggga tacgacgata
     6121 ccgaagacag ctcatgttat atcccgccgt taaccaccat caaacaggat tttcgcctgc
     6181 tggggcaaac cagcgtggac cgcttgctgc aactctctca gggccaggcg gtgaagggca
     6241 atcagctgtt gcccgtctca ctggtgaaaa gaaaaaccac cctggcgccc aatacgcaaa
     6301 cc
//
