LOCUS       sequence-267353-        7431 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Protein expression and purification of NR1D1_Hormone-recep.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7431)
  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 7431)
  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_NR1D1_Hormone-recep
FEATURES             Location/Qualifiers
     source          1..7431
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(116..133)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(207..229)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    218..234
                     /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)."
     promoter        complement(join(242..248,249..264,265..270))
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     CDS             complement(513..1595)
                     /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     1557..1576
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(1596..1673)
                     /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."
     primer_bind     complement(1696..1714)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1782..1886
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1887..1955,1956..2747)
                     /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(2105..2124)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2918..3505
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3659..3676
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3691..3831
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3757..3776)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3917..3939
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     rep_origin      3981..4436
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4068..4087)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4278..4299
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     terminator      complement(6029..6076)
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     CDS             complement(6818..6835)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             complement(6848..6871)
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             complement(join(6878..7431,1..100))
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     complement(6898..6920)
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
ORIGIN
        1 catacaaatg ctcttcatat ttttcttcaa gatattccaa aagaagtcga gtgggttgca
       61 caaggccctt aattttccaa taacctagta taggggacat aagcttaccg ctggaatggt
      121 gatggtgatg gtgttcttcg attttcataa tctatggtcc ttgttggtga agtgctcgtg
      181 aaaacaccta aacggactgg ctgtttcctg tgtgaaattg ttatccgctc acaattccac
      241 acattatacg agccgatgat taattgtcaa cagctcattt cagaatattt gccagaaccg
      301 ttatgatgtc ggcgcaaaaa acattatcca gaacgggagt gcgccttgag cgacacgaat
      361 tatgcagtga tttacgacct gcacagccat accacagctt ccgatggctg cctgacgcca
      421 gaagcattgg tgcaccgtgc agtcgatgat aagctgtcaa acatgagaat tgtgcctaat
      481 gagtgagcta acttacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
      541 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
      601 ggcgccaggg tggtttttct tttcaccagt gagacgggca acagctgatt gcccttcacc
      661 gcctggccct gagagagttg cagcaagcgg tccacgctgg tttgccccag caggcgaaaa
      721 tcctgtttga tggtggttaa cggcgggata taacatgagc tgtcttcggt atcgtcgtat
      781 cccactaccg agatatccgc accaacgcgc agcccggact cggtaatggc gcgcattgcg
      841 cccagcgcca tctgatcgtt ggcaaccagc atcgcagtgg gaacgatgcc ctcattcagc
      901 atttgcatgg tttgttgaaa accggacatg gcactccagt cgccttcccg ttccgctatc
      961 ggctgaattt gattgcgagt gagatattta tgccagccag ccagacgcag acgcgccgag
     1021 acagaactta atgggcccgc taacagcgcg atttgctggt gacccaatgc gaccagatgc
     1081 tccacgccca gtcgcgtacc gtcttcatgg gagaaaataa tactgttgat gggtgtctgg
     1141 tcagagacat caagaaataa cgccggaaca ttagtgcagg cagcttccac agcaatggca
     1201 tcctggtcat ccagcggata gttaatgatc agcccactga cgcgttgcgc gagaagattg
     1261 tgcaccgccg ctttacaggc ttcgacgccg cttcgttcta ccatcgacac caccacgctg
     1321 gcacccagtt gatcggcgcg agatttaatc gccgcgacaa tttgcgacgg cgcgtgcagg
     1381 gccagactgg aggtggcaac gccaatcagc aacgactgtt tgcccgccag ttgttgtgcc
     1441 acgcggttgg gaatgtaatt cagctccgcc atcgccgctt ccactttttc ccgcgttttc
     1501 gcagaaacgt ggctggcctg gttcaccacg cgggaaacgg tctgataaga gacaccggca
     1561 tactctgcga catcgtataa cgttactggt ttcacattca ccaccctgaa ttgactctct
     1621 tccgggcgct atcatgccat accgcgaaag gttttgcacc attcgatggt gtcgggaatt
     1681 cttgaagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa
     1741 tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     1801 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     1861 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     1921 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     1981 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     2041 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     2101 ttctgctatg tggcgcggta ttatcccgtg ttgacgccgg gcaagagcaa ctcggtcgcc
     2161 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     2221 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     2281 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     2341 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     2401 caaacgacga gcgtgacacc acgatgcctg cagcaatggc aacaacgttg cgcaaactat
     2461 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     2521 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     2581 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     2641 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     2701 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     2761 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     2821 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     2881 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     2941 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     3001 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     3061 ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     3121 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     3181 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     3241 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     3301 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     3361 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aaccctggta
     3421 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     3481 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     3541 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     3601 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag
     3661 cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg tattttctcc ttacgcatct
     3721 gtgcggtatt tcacaccgca tatggtgcac tctcagtaca atctgctctg atgccgcata
     3781 gttaagccag tatacactcc gctatcgcta cgtgactggg tcatggctgc gccccgacac
     3841 ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga
     3901 caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa
     3961 cgcgcgaggc aggatccgcg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     4021 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     4081 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     4141 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     4201 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     4261 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     4321 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     4381 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta caatttcgcg
     4441 gatcctgcgg taaagctcat cagcgtggtc gtgaagcgat tcacagatgt ctgcctgttc
     4501 atccgcgtcc agctcgttga gtttctccag aagcgttaat gtctggcttc tgataaagcg
     4561 ggccatgtta agggcggttt tttcctgttt ggtcactgat gcctccgtgt aagggggatt
     4621 tctgttcatg ggggtaatga taccgatgaa acgagagagg atgctcacga tacgggttac
     4681 tgatgatgaa catgcccggt tactggaacg ttgtgagggt aaacaactgg cggtatggat
     4741 gcggcgggac cagagaaaaa tcactcaggg tcaatgccag cgcttcgtta atacagatgt
     4801 aggtgttcca cagggtagcc agcagcatcc tgcgatgcag atccggaaca taatggtgca
     4861 gggcgctgac ttccgcgttt ccagacttta cgaaacacgg aaaccgaaga ccattcatgt
     4921 tgttgctcag gtcgcagacg ttttgcagca gcagtcgctt cacgttcgct cgcgtatcgg
     4981 tgattcattc tgctaaccag taaggcaacc ccgccagcct agccgggtcc tcaacgacag
     5041 gagcacgatc atgcgcaccc gtggccagga cccaacgctg cccgagatgc gccgcgtgcg
     5101 gctgctggag atggcggacg cgatggatat gttctgccaa gggttggttt gcgcattcac
     5161 agttctccgc aagaattgat tggctccaat tcttggagtg gtgaatccgt tagcgaggtg
     5221 ccgccggctt ccattcaggt cgaggtggcc cggctccatg caccgcgacg caacgcgggg
     5281 aggcagacaa ggtatagggc ggcgctagcc aaaagaactg gcatgattaa gactccttat
     5341 tacgcagtat gttagcaaac gtagaaaata catacataaa ggtggcaaca tataaaagaa
     5401 acgcaaagac accacggaat aagtttattt tgtcacaatc aatagaaaat tcatatggtt
     5461 taccagcgcc aaagacaaaa gggcgacatt caaccgattg agggagggaa ggtaaatatt
     5521 gacggaaatt attcattaaa ggtgaattat caccgtcacc gacttgagcc atttgggaat
     5581 tagagccagc aaaatcacca gtagcaccat taccattagc aaggccggaa acgtcaccaa
     5641 tgaaaccatc gatagcagca ccgtaatcag tagcgacaga atcaagtttg cctttagcgt
     5701 cagactgtag cgcgttttca tcggcatttt cggtcatagc ccccttatta gcgtttgcca
     5761 tcttttcata atcaaaatca ccggaaccag agccaccgcc actctagcta gacactgagt
     5821 ttcgtcacca gtacaaacta caacgcctgt agcattccac agacagccct catagttagc
     5881 gtaacgatct aaagttttgt cgtctttcca gacgttagta aatgaatttt ctgtatgggg
     5941 ttttgctaaa caactttcaa cagtttcagc ggagtgagaa tagaaaggaa caactaaagg
     6001 aattgcgaat aataattttt tcacgttgaa aatctccaaa aaaaaggctc caaaaggagc
     6061 ctttaattgt atcggtttat tatgcggccg cttgtgcatc aacacgaaaa gacagcagct
     6121 tttcgctatg catattattc agggtacgca gatccggcag cttcagcagc agtttggtaa
     6181 aacggctggt ttccagcgga cgatttttca gaaccagggc acgcagtgca cgcagcagtg
     6241 tctcttgcag ctgttcaacg cttgcgctat tttccatacc actacgatct gcgctaacca
     6301 gaacaactgc ggtaaacagt cccagttcct cttcggtcag ggccaggcta ttcagttttt
     6361 cactaaaatc aaacattgcg ctcagcagat cacccatacc cattgcaccc agttcttgca
     6421 ggctataggt ggtacggctc agaaacataa cggtctgatc tttaacatta aacaggctgg
     6481 caaaacgaac catcagaact tcaaaggtgc ctgctttcag cagggtaacc tgatcatgct
     6541 ggctcagatc acgaaaaccc ggaatgtgtt ttgcaaattc aacaacttca cgaactgccg
     6601 gtgtaaagct catgctaaaa tcttcccaaa tttcttgaac ggtacgaccg ctacgaccat
     6661 gcggatacgg gctaccgctt gcatgatttg cattaaagtt acccggactg ctacccagtt
     6721 tatcatgtgc ataggtaaag atttcacgat gggcacgtgc aacctggcta ataacatctt
     6781 caacggtcgg ttcaggggcc ggctgggccg cggtaccgga accacgcgga accagaccgc
     6841 tggacagggg cccctggaac agaacttcca gatccgattt tggaggatgg tcgccaccac
     6901 caaacgtggc ttgccagccc tgcaaaggcc atgctatata cttgctggat ttcaagtact
     6961 tatcaatttg tgggatagct tcaatacgtt ttttaaaaca aactaatttt gggaacgcat
     7021 ccaggcacat tgggtccatg tataaaacaa catcaagagc gtcatacaac atgaagtcag
     7081 gatgggttac atgatcacca tttaaatatg ttttatgaca taaacgatct tcgaacattt
     7141 tcagcatttc aggtagcttg ctaagaaaat caactttgag agtttcaaag tctttactat
     7201 atgcaattct cgaaacaccg tatctaatat ccaaaaccgc tccttcaagc attgaaatct
     7261 ctgcacgctc ttttggacaa ccacccaaca tgttgtgctt gtcagctata taacgtatga
     7321 tggccataga ctgtgttaat ttaacatcac catcaatata ataaggaaga ttgggaaact
     7381 ccaaacccaa ttcaaacttt ttgtttcgcc atttatcacc ttcatcgcgc t
//
