LOCUS       sequence-320650-        6888 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expressing mouse Hgd in bacterial cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6888)
  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 6888)
  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: pHis-Hgd
FEATURES             Location/Qualifiers
     source          1..6888
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      28..483
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(115..134)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     325..346
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        509..613
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(614..682,683..1474)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(832..851)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1645..2233
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2134..2153
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2387..2404
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2419..2558
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2484..2503)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2644..2666
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2660..2851)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(3660..4742)
                     /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     4704..4723
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4743..4820)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5026..5045
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     5129..5148
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5129..5147
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5148..5172
                     /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)."
     RBS             5187..5209
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             5229..5246
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             5268..5288
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     RBS             6296..6304
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             6748..6765
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6818..6836)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      6832..6879
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 ggaactatat ccggattggc gaatgggacg cgccctgtag cggcgcatta agcgcggcgg
       61 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
      121 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
      181 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
      241 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
      301 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
      361 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
      421 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa
      481 tttaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
      541 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
      601 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
      661 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
      721 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
      781 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
      841 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
      901 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
      961 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     1021 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     1081 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     1141 tgacaccacg atgcctgcag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     1201 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     1261 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     1321 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     1381 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     1441 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     1501 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     1561 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     1621 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     1681 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     1741 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt
     1801 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     1861 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     1921 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     1981 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     2041 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     2101 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     2161 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     2221 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     2281 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     2341 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     2401 cgaggaagcg gaagagcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc
     2461 acaccgcaat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat
     2521 acactccgct atcgctacgt gactgggtca tggctgcgcc ccgacacccg ccaacacccg
     2581 ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg
     2641 tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgaggcagc
     2701 tgcggtaaag ctcatcagcg tggtcgtgaa gcgattcaca gatgtctgcc tgttcatccg
     2761 cgtccagctc gttgagtttc tccagaagcg ttaatgtctg gcttctgata aagcgggcca
     2821 tgttaagggc ggttttttcc tgtttggtca ctgatgcctc cgtgtaaggg ggatttctgt
     2881 tcatgggggt aatgataccg atgaaacgag agaggatgct cacgatacgg gttactgatg
     2941 atgaacatgc ccggttactg gaacgttgtg agggtaaaca actggcggta tggatgcggc
     3001 gggaccagag aaaaatcact cagggtcaat gccagcgctt cgttaataca gatgtaggtg
     3061 ttccacaggg tagccagcag catcctgcga tgcagatccg gaacataatg gtgcagggcg
     3121 ctgacttccg cgtttccaga ctttacgaaa cacggaaacc gaagaccatt catgttgttg
     3181 ctcaggtcgc agacgttttg cagcagcagt cgcttcacgt tcgctcgcgt atcggtgatt
     3241 cattctgcta accagtaagg caaccccgcc agcctagccg ggtcctcaac gacaggagca
     3301 cgatcatgcg cacccgtggg gccgccatgc cggcgataat ggcctgcttc tcgccgaaac
     3361 gtttggtggc gggaccagtg acgaaggctt gagcgagggc gtgcaagatt ccgaataccg
     3421 caagcgacag gccgatcatc gtcgcgctcc agcgaaagcg gtcctcgccg aaaatgaccc
     3481 agagcgctgc cggcacctgt cctacgagtt gcatgataaa gaagacagtc ataagtgcgg
     3541 cgacgatagt catgccccgc gcccaccgga aggagctgac tgggttgaag gctctcaagg
     3601 gcatcggtcg agatcccggt gcctaatgag tgagctaact tacattaatt gcgttgcgct
     3661 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     3721 gcgcggggag aggcggtttg cgtattgggc gccagggtgg tttttctttt caccagtgag
     3781 acgggcaaca gctgattgcc cttcaccgcc tggccctgag agagttgcag caagcggtcc
     3841 acgctggttt gccccagcag gcgaaaatcc tgtttgatgg tggttaacgg cgggatataa
     3901 catgagctgt cttcggtatc gtcgtatccc actaccgaga tatccgcacc aacgcgcagc
     3961 ccggactcgg taatggcgcg cattgcgccc agcgccatct gatcgttggc aaccagcatc
     4021 gcagtgggaa cgatgccctc attcagcatt tgcatggttt gttgaaaacc ggacatggca
     4081 ctccagtcgc cttcccgttc cgctatcggc tgaatttgat tgcgagtgag atatttatgc
     4141 cagccagcca gacgcagacg cgccgagaca gaacttaatg ggcccgctaa cagcgcgatt
     4201 tgctggtgac ccaatgcgac cagatgctcc acgcccagtc gcgtaccgtc ttcatgggag
     4261 aaaataatac tgttgatggg tgtctggtca gagacatcaa gaaataacgc cggaacatta
     4321 gtgcaggcag cttccacagc aatggcatcc tggtcatcca gcggatagtt aatgatcagc
     4381 ccactgacgc gttgcgcgag aagattgtgc accgccgctt tacaggcttc gacgccgctt
     4441 cgttctacca tcgacaccac cacgctggca cccagttgat cggcgcgaga tttaatcgcc
     4501 gcgacaattt gcgacggcgc gtgcagggcc agactggagg tggcaacgcc aatcagcaac
     4561 gactgtttgc ccgccagttg ttgtgccacg cggttgggaa tgtaattcag ctccgccatc
     4621 gccgcttcca ctttttcccg cgttttcgca gaaacgtggc tggcctggtt caccacgcgg
     4681 gaaacggtct gataagagac accggcatac tctgcgacat cgtataacgt tactggtttc
     4741 acattcacca ccctgaattg actctcttcc gggcgctatc atgccatacc gcgaaaggtt
     4801 ttgcgccatt cgatggtgtc cgggatctcg acgctctccc ttatgcgact cctgcattag
     4861 gaagcagccc agtagtaggt tgaggccgtt gagcaccgcc gccgcaagga atggtgcatg
     4921 caaggagatg gcgcccaaca gtcccccggc cacggggcct gccaccatac ccacgccgaa
     4981 acaagcgctc atgagcccga agtggcgagc ccgatcttcc ccatcggtga tgtcggcgat
     5041 ataggcgcca gcaaccgcac ctgtggcgcc ggtgatgccg gccacgatgc gtccggcgta
     5101 gaggatcgag atctcgatcc cgcgaaatta atacgactca ctatagggga attgtgagcg
     5161 gataacaatt cccctctaga aataattttg tttaacttta agaaggagat atacatatgt
     5221 cgtactacca tcaccatcac catcacgatt acgatatccc aacgaccgaa aacctgtatt
     5281 ttcagggcgc catggatccg gaattcatgg cagagttaaa gtacatttct ggatttggga
     5341 atgagtgtgc ttcagaggat cctcgctgtc caggttccct gcccaaagga cagaataatc
     5401 ctcaggtctg cccctacaac ctctacgctg agcagctctc gggatcagcc ttcacttgtc
     5461 cacggaacac caataagaga agctggctgt ataggattct tccttcagtt tctcacaaac
     5521 cctttgagtc catcgaccaa ggccatgtca ctcacaactg ggatgaagtt ggccctgatc
     5581 ctaaccagct aagatggaaa ccatttgaga ttccaaaagc gtctgagaag aaagtggact
     5641 ttgtgagtgg tttgtacacc ttatgtggag caggagacat caagtctaac aacggccttg
     5701 ctgtgcacat ttttctgtgt aactcctcca tggagaacag gtgcttttac aactcagatg
     5761 gggatttcct gattgttccc cagaaaggga aacttctcat ttacaccgag tttggcaaga
     5821 tgtccctgca gcccaatgaa atctgcgtca ttcagagagg aatgcgattc agcgtcgatg
     5881 tctttgagga gaccaggggc tacatcctgg aggtctatgg tgtccacttt gagttacctg
     5941 acctgggacc catcggagca aatggcttgg ccaatcctcg agatttcttg attcccgttg
     6001 cctggtatga agatcgtcga gtgccaggtg gttacactgt gattaataaa ttccagggca
     6061 agctgtttgc ttgcaaacag gatgtctctc cattcaatgt cgtggcctgg catggaaact
     6121 acacacccta caagtacaac ctagagaact tcatggtcat caatgcggta gcctttgacc
     6181 atgcggatcc ttccattttc accgtgctga ctgccaaatc cctccgacct ggggtggcta
     6241 ttgctgactt tgtcatcttc ccacctcggt ggggagttgc agataaaacc ttcagacctc
     6301 cttattacca caggaactgc atgagtgagt tcatgggcct catcaaaggt cactatgagg
     6361 caaagcaagg tggattcctg ccagggggcg gcagcctgca cagtgccatg acccctcacg
     6421 gccccgatgc agactgcttt gaaaaggcca gcaaggccaa actcgagcca gagaggatag
     6481 ctgacggcac catggcgttt atgtttgaat cttccttaag tttggcggtc accaagtggg
     6541 gactcaagac ctgtagctgt ttggatgaga actactacaa atgttgggag cctctccgga
     6601 gccacttcac ccccaattcc aggagcccga cagagcctaa gtgaaattca aaggcctacg
     6661 tcgacgagct caactagtgc ggccgctttc gaatctagag cctgcagtct cgaggcatgc
     6721 ggtaccaagc ttgcggccgc actcgagcac caccaccacc accactgaga tccggctgct
     6781 aacaaagccc gaaaggaagc tgagttggct gctgccaccg ctgagcaata actagcataa
     6841 ccccttgggg cctctaaacg ggtcttgagg ggttttttgc tgaaagga
//
