LOCUS       sequence-320646-        5693 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expressing mouse Gstz1 in bacterial cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5693)
  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 5693)
  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-GstZ1
FEATURES             Location/Qualifiers
     source          1..5693
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(86..91,92..107,108..114)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    122..138
                     /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             161..814
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     772..794
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             827..847
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(1646..1668)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1813..1831)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1899..2003
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2004..2072,2073..2864)
                     /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(2222..2241)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3035..3623
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3524..3543
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3777..3794
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        3867..3944
                     /gene="lacI"
                     /label=lacI promoter
     CDS             3945..5027
                     /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(3964..3983)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(5076..5081,5082..5099,5100..5106)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5114..5130
                     /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     5138..5154
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5138..5154
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             5150..5677
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(5170..5187)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5170..5186)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5179..5201)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 tgtcgctcaa ggcgcactcc cgttctggat aatgtttttt gcgccgacat cataacggtt
       61 ctggcaaata ttctgaaatg agctgttgac aattaatcat cggctcgtat aatgtgtgga
      121 attgtgagcg gataacaatt tcacacagga aacagtattc atgtccccta tactaggtta
      181 ttggaaaatt aagggccttg tgcaacccac tcgacttctt ttggaatatc ttgaagaaaa
      241 atatgaagag catttgtatg agcgcgatga aggtgataaa tggcgaaaca aaaagtttga
      301 attgggtttg gagtttccca atcttcctta ttatattgat ggtgatgtta aattaacaca
      361 gtctatggcc atcatacgtt atatagctga caagcacaac atgttgggtg gttgtccaaa
      421 agagcgtgca gagatttcaa tgcttgaagg agcggttttg gatattagat acggtgtttc
      481 gagaattgca tatagtaaag actttgaaac tctcaaagtt gattttctta gcaagctacc
      541 tgaaatgctg aaaatgttcg aagatcgttt atgtcataaa acatatttaa atggtgatca
      601 tgtaacccat cctgacttca tgttgtatga cgctcttgat gttgttttat acatggaccc
      661 aatgtgcctg gatgcgttcc caaaattagt ttgttttaaa aaacgtattg aagctatccc
      721 acaaattgat aagtacttga aatccagcaa gtatatagca tggcctttgc agggctggca
      781 agccacgttt ggtggtggcg accatcctcc aaaaatcgat acgaccgaaa acctgtattt
      841 tcagggcgcc atggatccgg aattcatgca agccgggaag cctatcctct actcctattt
      901 ccgaagctcc tgttcatgga gagttcgaat tgctctggcg ttaaaaggca ttgactatga
      961 gatagtgccc atcaacctca taaaggatgg cgggcaacag ttcactgagg aatttcagac
     1021 cctgaacccc atgaagcaag tgccagcgct gaaaattgat ggaatcacca ttgtccagtc
     1081 actggccatc atggagtacc tggaagagac tcggcctatc ccacggctcc tgcctcagga
     1141 cccacagaaa agagccatcg tgcgcatgat ttctgacctc atcgctagtg gcatccagcc
     1201 ccttcagaac ctgtctgtcc tgaagcaagt gggacaggag aaccagatgc aatgggccca
     1261 aaaggttatt acttccggct ttaacgctct ggagaagatt ctgcagagca cagcagggaa
     1321 gtactgcgta ggagatgagg tatccatggc tgatgtgtgc ttagtacccc aagtggcaaa
     1381 tgctgaaagg ttcaaggtgg atctaagccc ctaccctacc atcagtcaca tcaacaagga
     1441 gctgctagcc ttagaggttt tccaagtgtc tcacccccgt cgacagccag atacccctgc
     1501 cgagctgagg acttagaatt caaaggccta cgtcgacgag ctcaactagt gcggccgctt
     1561 tcgaatctag agcctgcagt ctcgagcggc cgcatcgtga ctgactgacg atctgcctcg
     1621 cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag
     1681 cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg
     1741 gcgggtgtcg gggcgcagcc atgacccagt cacgtagcga tagcggagtg tataattctt
     1801 gaagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg
     1861 tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     1921 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     1981 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     2041 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     2101 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     2161 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     2221 tgctatgtgg cgcggtatta tcccgtgttg acgccgggca agagcaactc ggtcgccgca
     2281 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     2341 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     2401 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     2461 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     2521 acgacgagcg tgacaccacg atgcctgcag caatggcaac aacgttgcgc aaactattaa
     2581 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     2641 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     2701 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     2761 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     2821 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     2881 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     2941 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     3001 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     3061 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     3121 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     3181 tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     3241 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     3301 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     3361 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     3421 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     3481 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     3541 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     3601 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     3661 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     3721 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
     3781 agtcagtgag cgaggaagcg gaagagcgcc tgatgcggta ttttctcctt acgcatctgt
     3841 gcggtatttc acaccgcata aattccgaca ccatcgaatg gcgcaaaacc tttcgcggta
     3901 tggcatgata gcgcccggaa gagagtcaat tcagggtggt gaatgtgaaa ccagtaacgt
     3961 tatacgatgt cgcagagtat gccggtgtct cttatcagac cgtttcccgc gtggtgaacc
     4021 aggccagcca cgtttctgcg aaaacgcggg aaaaagtgga agcggcgatg gcggagctga
     4081 attacattcc caaccgcgtg gcacaacaac tggcgggcaa acagtcgttg ctgattggcg
     4141 ttgccacctc cagtctggcc ctgcacgcgc cgtcgcaaat tgtcgcggcg attaaatctc
     4201 gcgccgatca actgggtgcc agcgtggtgg tgtcgatggt agaacgaagc ggcgtcgaag
     4261 cctgtaaagc ggcggtgcac aatcttctcg cgcaacgcgt cagtgggctg atcattaact
     4321 atccgctgga tgaccaggat gccattgctg tggaagctgc ctgcactaat gttccggcgt
     4381 tatttcttga tgtctctgac cagacaccca tcaacagtat tattttctcc catgaagacg
     4441 gtacgcgact gggcgtggag catctggtcg cattgggtca ccagcaaatc gcgctgttag
     4501 cgggcccatt aagttctgtc tcggcgcgtc tgcgtctggc tggctggcat aaatatctca
     4561 ctcgcaatca aattcagccg atagcggaac gggaaggcga ctggagtgcc atgtccggtt
     4621 ttcaacaaac catgcaaatg ctgaatgagg gcatcgttcc cactgcgatg ctggttgcca
     4681 acgatcagat ggcgctgggc gcaatgcgcg ccattaccga gtccgggctg cgcgttggtg
     4741 cggatatctc ggtagtggga tacgacgata ccgaagacag ctcatgttat atcccgccgt
     4801 taaccaccat caaacaggat tttcgcctgc tggggcaaac cagcgtggac cgcttgctgc
     4861 aactctctca gggccaggcg gtgaagggca atcagctgtt gcccgtctca ctggtgaaaa
     4921 gaaaaaccac cctggcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat
     4981 taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt
     5041 aatgtaagtt agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt
     5101 atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat
     5161 tacggattca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca
     5221 acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg
     5281 caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatggcgct ttgcctggtt
     5341 tccggcacca gaagcggtgc cggaaagctg gctggagtgc gatcttcctg aggccgatac
     5401 tgtcgtcgtc ccctcaaact ggcagatgca cggttacgat gcgcccatct acaccaacgt
     5461 gacctatccc attacggtca atccgccgtt tgttcccacg gagaatccga cgggttgtta
     5521 ctcgctcaca tttaatgttg atgaaagctg gctacaggaa ggccagacgc gaattatttt
     5581 tgatggcgtt ggaattagct tatcgactgc acggtgcacc aatgcttctg gcgtcaggca
     5641 gccatcggaa gctgtggtat ggctgtgcag gtcgtaaatc actgcataat tcg
//
