LOCUS       sequence-215199-        7458 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Protein expression and purification of NR1H3_Hormone-recep.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7458)
  AUTHORS   Teyra J, Kelil A, Jain S, Helmy M, Jajodia R, Hooda Y, Gu J, D'Cruz 
            AA, Nicholson SE, Min J, Sudol M, Kim PM, Bader GD, Sidhu SS
  TITLE     Large-scale survey and database of high affinity ligands for peptide
            recognition modules.
  JOURNAL   Mol Syst Biol. 2020 Dec;16(12):e9310. doi: 10.15252/msb.20199310.
  PUBMED    33438817
REFERENCE   2  (bases 1 to 7458)
  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"; journalName: "Mol Syst 
            Biol."; date: "2020-12"; volume: "16(12):e9310. doi"; pages: " 
            10.15252/msb.20199310"
COMMENT     Sequence Label: pHH0103_NR1H3_Hormone-recep
FEATURES             Location/Qualifiers
     source          1..7458
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(1..6,7..22,23..29)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    37..53
                     /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     42..64
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     CDS             138..155
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             171..824
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     782..804
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             831..854
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             867..884
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     terminator      1653..1700
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      complement(3293..3748)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3430..3451)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3642..3661
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     complement(3790..3812)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3898..4038
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3953..3972
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4053..4070)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4224..4811)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(4982..5773,5774..5842))
                     /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     5605..5624
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5843..5947)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6015..6033
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6056..6133
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             6134..7216
                     /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(6153..6172)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 ttgacaatta atcatcggct cgtataatgt gtggaattgt gagcggataa caatttcaca
       61 caggaaacag ccagtccgtt taggtgtttt cacgagcact tcaccaacaa ggaccataga
      121 ttatgaaaat cgaagaacac catcaccatc accattccag cggtaagctt atgtccccta
      181 tactaggtta ttggaaaatt aagggccttg tgcaacccac tcgacttctt ttggaatatc
      241 ttgaagaaaa atatgaagag catttgtatg agcgcgatga aggtgataaa tggcgaaaca
      301 aaaagtttga attgggtttg gagtttccca atcttcctta ttatattgat ggtgatgtta
      361 aattaacaca gtctatggcc atcatacgtt atatagctga caagcacaac atgttgggtg
      421 gttgtccaaa agagcgtgca gagatttcaa tgcttgaagg agcggttttg gatattagat
      481 acggtgtttc gagaattgca tatagtaaag actttgaaac tctcaaagtt gattttctta
      541 gcaagctacc tgaaatgctg aaaatgttcg aagatcgttt atgtcataaa acatatttaa
      601 atggtgatca tgtaacccat cctgacttca tgttgtatga cgctcttgat gttgttttat
      661 acatggaccc aatgtgcctg gatgcgttcc caaaattagt ttgttttaaa aaacgtattg
      721 aagctatccc acaaattgat aagtacttga aatccagcaa gtatatagca tggcctttgc
      781 agggctggca agccacgttt ggtggtggcg accatcctcc aaaatcggat ctggaagttc
      841 tgttccaggg gcccctgtcc agcggtctgg ttccgcgtgg ttccggtacc gcggcccagc
      901 cggcccctca gctgtcacct gaacagctgg gtatgattga aaaactggtt gcagcacagc
      961 agcagtgtaa tcgtcgtagc tttagcgatc gtctgcgtgt taccccgtgg ccgattgcac
     1021 cggatccgca gagccgtgaa gcacgtcagc agcgttttgc acactttacc gaactggcaa
     1081 ttgttagcgt tcaagagatt gtggatttcg ccaaacagct gcctggtttt ctgcagctga
     1141 gtcgtgaaga tcagattgcc ctgctgaaaa cctcagcaat tgaagttatg ctgctggaaa
     1201 ccagccgtcg ttataatccg ggtagcgaaa gcattacctt tctgaaagat tttagctata
     1261 accgcgagga ttttgcgaaa gccggtctgc aggttgaatt tatcaatccg atctttgaat
     1321 tcagccgtgc catgaatgaa ctgcaactga atgatgcaga atttgcactg ctgattgcca
     1381 ttagcatttt tagcgcagat cgtccgaatg ttcaggatca gttacaggtg gaacgtctgc
     1441 agcataccta tgttgaagca ctgcatgcat atgtgagcat taatcatccg catgatccgc
     1501 tgatgtttcc gcgtatgctg atgaaactgg tgagcctgcg taccctgagc agcgtgcata
     1561 gcgaacaggt ttttgcgctg cgcctgcagg ataaaaaact gcctccgctg ctgtcagaaa
     1621 tttgggatgt tcatgaagcg gccgcataat aaaccgatac aattaaaggc tccttttgga
     1681 gccttttttt tggagatttt caacgtgaaa aaattattat tcgcaattcc tttagttgtt
     1741 cctttctatt ctcactccgc tgaaactgtt gaaagttgtt tagcaaaacc ccatacagaa
     1801 aattcattta ctaacgtctg gaaagacgac aaaactttag atcgttacgc taactatgag
     1861 ggctgtctgt ggaatgctac aggcgttgta gtttgtactg gtgacgaaac tcagtgtcta
     1921 gctagagtgg cggtggctct ggttccggtg attttgatta tgaaaagatg gcaaacgcta
     1981 ataagggggc tatgaccgaa aatgccgatg aaaacgcgct acagtctgac gctaaaggca
     2041 aacttgattc tgtcgctact gattacggtg ctgctatcga tggtttcatt ggtgacgttt
     2101 ccggccttgc taatggtaat ggtgctactg gtgattttgc tggctctaat tcccaaatgg
     2161 ctcaagtcgg tgacggtgat aattcacctt taatgaataa tttccgtcaa tatttacctt
     2221 ccctccctca atcggttgaa tgtcgccctt ttgtctttgg cgctggtaaa ccatatgaat
     2281 tttctattga ttgtgacaaa ataaacttat tccgtggtgt ctttgcgttt cttttatatg
     2341 ttgccacctt tatgtatgta ttttctacgt ttgctaacat actgcgtaat aaggagtctt
     2401 aatcatgcca gttcttttgg ctagcgccgc cctatacctt gtctgcctcc ccgcgttgcg
     2461 tcgcggtgca tggagccggg ccacctcgac ctgaatggaa gccggcggca cctcgctaac
     2521 ggattcacca ctccaagaat tggagccaat caattcttgc ggagaactgt gaatgcgcaa
     2581 accaaccctt ggcagaacat atccatcgcg tccgccatct ccagcagccg cacgcggcgc
     2641 atctcgggca gcgttgggtc ctggccacgg gtgcgcatga tcgtgctcct gtcgttgagg
     2701 acccggctag gctggcgggg ttgccttact ggttagcaga atgaatcacc gatacgcgag
     2761 cgaacgtgaa gcgactgctg ctgcaaaacg tctgcgacct gagcaacaac atgaatggtc
     2821 ttcggtttcc gtgtttcgta aagtctggaa acgcggaagt cagcgccctg caccattatg
     2881 ttccggatct gcatcgcagg atgctgctgg ctaccctgtg gaacacctac atctgtatta
     2941 acgaagcgct ggcattgacc ctgagtgatt tttctctggt cccgccgcat ccataccgcc
     3001 agttgtttac cctcacaacg ttccagtaac cgggcatgtt catcatcagt aacccgtatc
     3061 gtgagcatcc tctctcgttt catcggtatc attaccccca tgaacagaaa tcccccttac
     3121 acggaggcat cagtgaccaa acaggaaaaa accgccctta acatggcccg ctttatcaga
     3181 agccagacat taacgcttct ggagaaactc aacgagctgg acgcggatga acaggcagac
     3241 atctgtgaat cgcttcacga ccacgctgat gagctttacc gcaggatccg cgaaattgta
     3301 agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac
     3361 caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg
     3421 agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa
     3481 gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt
     3541 tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt
     3601 agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga
     3661 gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc
     3721 gcgcttaatg cgccgctaca gggcgcgtcg cggatcctgc ctcgcgcgtt tcggtgatga
     3781 cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga
     3841 tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggcgc
     3901 agccatgacc cagtcacgta gcgatagcgg agtgtatact ggcttaacta tgcggcatca
     3961 gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg
     4021 agaaaatacc gcatcaggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     4081 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     4141 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     4201 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     4261 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccagggtttc
     4321 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     4381 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     4441 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     4501 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     4561 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     4621 cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct
     4681 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     4741 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     4801 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     4861 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     4921 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     4981 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     5041 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     5101 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     5161 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     5221 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     5281 acgttgttgc cattgctgca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     5341 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     5401 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     5461 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     5521 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     5581 gctcttgccc ggcgtcaaca cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     5641 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     5701 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     5761 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     5821 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     5881 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     5941 ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga
     6001 cattaaccta taaaaatagg cgtatcacga ggccctttcg tcttcaagaa ttcccgacac
     6061 catcgaatgg tgcaaaacct ttcgcggtat ggcatgatag cgcccggaag agagtcaatt
     6121 cagggtggtg aatgtgaaac cagtaacgtt atacgatgtc gcagagtatg ccggtgtctc
     6181 ttatcagacc gtttcccgcg tggtgaacca ggccagccac gtttctgcga aaacgcggga
     6241 aaaagtggaa gcggcgatgg cggagctgaa ttacattccc aaccgcgtgg cacaacaact
     6301 ggcgggcaaa cagtcgttgc tgattggcgt tgccacctcc agtctggccc tgcacgcgcc
     6361 gtcgcaaatt gtcgcggcga ttaaatctcg cgccgatcaa ctgggtgcca gcgtggtggt
     6421 gtcgatggta gaacgaagcg gcgtcgaagc ctgtaaagcg gcggtgcaca atcttctcgc
     6481 gcaacgcgtc agtgggctga tcattaacta tccgctggat gaccaggatg ccattgctgt
     6541 ggaagctgcc tgcactaatg ttccggcgtt atttcttgat gtctctgacc agacacccat
     6601 caacagtatt attttctccc atgaagacgg tacgcgactg ggcgtggagc atctggtcgc
     6661 attgggtcac cagcaaatcg cgctgttagc gggcccatta agttctgtct cggcgcgtct
     6721 gcgtctggct ggctggcata aatatctcac tcgcaatcaa attcagccga tagcggaacg
     6781 ggaaggcgac tggagtgcca tgtccggttt tcaacaaacc atgcaaatgc tgaatgaggg
     6841 catcgttccc actgcgatgc tggttgccaa cgatcagatg gcgctgggcg caatgcgcgc
     6901 cattaccgag tccgggctgc gcgttggtgc ggatatctcg gtagtgggat acgacgatac
     6961 cgaagacagc tcatgttata tcccgccgtt aaccaccatc aaacaggatt ttcgcctgct
     7021 ggggcaaacc agcgtggacc gcttgctgca actctctcag ggccaggcgg tgaagggcaa
     7081 tcagctgttg cccgtctcac tggtgaaaag aaaaaccacc ctggcgccca atacgcaaac
     7141 cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact
     7201 ggaaagcggg cagtgagcgc aacgcaatta atgtaagtta gctcactcat taggcacaat
     7261 tctcatgttt gacagcttat catcgactgc acggtgcacc aatgcttctg gcgtcaggca
     7321 gccatcggaa gctgtggtat ggctgtgcag gtcgtaaatc actgcataat tcgtgtcgct
     7381 caaggcgcac tcccgttctg gataatgttt tttgcgccga catcataacg gttctggcaa
     7441 atattctgaa atgagctg
//
