LOCUS       sequence-217293-        7542 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Protein expression and purification of NR4A2_Hormone-recep.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7542)
  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 7542)
  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_NR4A2_Hormone-recep
FEATURES             Location/Qualifiers
     source          1..7542
                     /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      1737..1784
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      complement(3377..3832)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3514..3535)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3726..3745
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     complement(3874..3896)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3982..4122
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4037..4056
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4137..4154)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4308..4895)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(5066..5857,5858..5926))
                     /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     5689..5708
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5927..6031)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6099..6117
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6140..6217
                     /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             6218..7300
                     /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(6237..6256)
                     /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 cggccggtat ggttaaagaa gttgttcgta ccgatagcct gaaaggtcgt cgtggtcgtc
      961 tgccgagcaa accgaaaagt ccgcaagaac cgagccctcc gagcccaccg gttagcctga
     1021 ttagcgcact ggttcgtgca catgttgata gcaatccggc aatgaccagc ctggattata
     1081 gccgttttca ggcaaacccg gattatcaga tgagcggtga tgatacccag catattcagc
     1141 agttttatga tctgctgacc ggtagcatgg aaattattcg tggttgggca gaaaaaattc
     1201 ctggttttgc cgatctgccg aaagccgatc aggacctgct gtttgaaagc gcatttctgg
     1261 aactgtttgt tctgcgtctg gcatatcgta gtaatccggt tgaaggtaaa ctgatttttt
     1321 gcaatggtgt tgtgctgcat cgtctgcagt gtgtgcgtgg ttttggtgaa tggattgatt
     1381 ccattgttga gtttagcagc aatctgcaga acatgaacat tgatattagc gcctttagct
     1441 gtattgcagc actggcaatg gttaccgaac gtcatggtct gaaagaacct aaacgtgttg
     1501 aagaactgca gaataaaatc gtgaactgcc tgaaggatca tgtgaccttt aataacggtg
     1561 gcctgaatcg tccgaattat ctgagcaaac tgttaggcaa actgccggaa ctgcgtaccc
     1621 tgtgcaccca gggtctgcag cgtatctttt atctgaaact ggaagatctg gttccgccac
     1681 cggcaattat tgataaactg tttctggata ccctgccgtt tgcggccgca taataaaccg
     1741 atacaattaa aggctccttt tggagccttt tttttggaga ttttcaacgt gaaaaaatta
     1801 ttattcgcaa ttcctttagt tgttcctttc tattctcact ccgctgaaac tgttgaaagt
     1861 tgtttagcaa aaccccatac agaaaattca tttactaacg tctggaaaga cgacaaaact
     1921 ttagatcgtt acgctaacta tgagggctgt ctgtggaatg ctacaggcgt tgtagtttgt
     1981 actggtgacg aaactcagtg tctagctaga gtggcggtgg ctctggttcc ggtgattttg
     2041 attatgaaaa gatggcaaac gctaataagg gggctatgac cgaaaatgcc gatgaaaacg
     2101 cgctacagtc tgacgctaaa ggcaaacttg attctgtcgc tactgattac ggtgctgcta
     2161 tcgatggttt cattggtgac gtttccggcc ttgctaatgg taatggtgct actggtgatt
     2221 ttgctggctc taattcccaa atggctcaag tcggtgacgg tgataattca cctttaatga
     2281 ataatttccg tcaatattta ccttccctcc ctcaatcggt tgaatgtcgc ccttttgtct
     2341 ttggcgctgg taaaccatat gaattttcta ttgattgtga caaaataaac ttattccgtg
     2401 gtgtctttgc gtttctttta tatgttgcca cctttatgta tgtattttct acgtttgcta
     2461 acatactgcg taataaggag tcttaatcat gccagttctt ttggctagcg ccgccctata
     2521 ccttgtctgc ctccccgcgt tgcgtcgcgg tgcatggagc cgggccacct cgacctgaat
     2581 ggaagccggc ggcacctcgc taacggattc accactccaa gaattggagc caatcaattc
     2641 ttgcggagaa ctgtgaatgc gcaaaccaac ccttggcaga acatatccat cgcgtccgcc
     2701 atctccagca gccgcacgcg gcgcatctcg ggcagcgttg ggtcctggcc acgggtgcgc
     2761 atgatcgtgc tcctgtcgtt gaggacccgg ctaggctggc ggggttgcct tactggttag
     2821 cagaatgaat caccgatacg cgagcgaacg tgaagcgact gctgctgcaa aacgtctgcg
     2881 acctgagcaa caacatgaat ggtcttcggt ttccgtgttt cgtaaagtct ggaaacgcgg
     2941 aagtcagcgc cctgcaccat tatgttccgg atctgcatcg caggatgctg ctggctaccc
     3001 tgtggaacac ctacatctgt attaacgaag cgctggcatt gaccctgagt gatttttctc
     3061 tggtcccgcc gcatccatac cgccagttgt ttaccctcac aacgttccag taaccgggca
     3121 tgttcatcat cagtaacccg tatcgtgagc atcctctctc gtttcatcgg tatcattacc
     3181 cccatgaaca gaaatccccc ttacacggag gcatcagtga ccaaacagga aaaaaccgcc
     3241 cttaacatgg cccgctttat cagaagccag acattaacgc ttctggagaa actcaacgag
     3301 ctggacgcgg atgaacaggc agacatctgt gaatcgcttc acgaccacgc tgatgagctt
     3361 taccgcagga tccgcgaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt
     3421 tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca
     3481 aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta
     3541 aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta
     3601 cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg
     3661 aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga
     3721 aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg
     3781 ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcgcggatc
     3841 ctgcctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac
     3901 ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc
     3961 gggtgttggc gggtgtcggg gcgcagccat gacccagtca cgtagcgata gcggagtgta
     4021 tactggctta actatgcggc atcagagcag attgtactga gagtgcacca tatgcggtgt
     4081 gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgctcttc cgcttcctcg
     4141 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     4201 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     4261 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     4321 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     4381 ggactataaa gataccaggg tttccccctg gaagctccct cgtgcgctct cctgttccga
     4441 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     4501 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     4561 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     4621 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     4681 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     4741 ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     4801 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     4861 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     4921 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     4981 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     5041 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     5101 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     5161 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     5221 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     5281 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     5341 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tgcaggcatc gtggtgtcac
     5401 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     5461 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     5521 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     5581 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     5641 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aacacgggat aataccgcgc
     5701 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     5761 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     5821 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     5881 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     5941 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     6001 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     6061 tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct
     6121 ttcgtcttca agaattcccg acaccatcga atggtgcaaa acctttcgcg gtatggcatg
     6181 atagcgcccg gaagagagtc aattcagggt ggtgaatgtg aaaccagtaa cgttatacga
     6241 tgtcgcagag tatgccggtg tctcttatca gaccgtttcc cgcgtggtga accaggccag
     6301 ccacgtttct gcgaaaacgc gggaaaaagt ggaagcggcg atggcggagc tgaattacat
     6361 tcccaaccgc gtggcacaac aactggcggg caaacagtcg ttgctgattg gcgttgccac
     6421 ctccagtctg gccctgcacg cgccgtcgca aattgtcgcg gcgattaaat ctcgcgccga
     6481 tcaactgggt gccagcgtgg tggtgtcgat ggtagaacga agcggcgtcg aagcctgtaa
     6541 agcggcggtg cacaatcttc tcgcgcaacg cgtcagtggg ctgatcatta actatccgct
     6601 ggatgaccag gatgccattg ctgtggaagc tgcctgcact aatgttccgg cgttatttct
     6661 tgatgtctct gaccagacac ccatcaacag tattattttc tcccatgaag acggtacgcg
     6721 actgggcgtg gagcatctgg tcgcattggg tcaccagcaa atcgcgctgt tagcgggccc
     6781 attaagttct gtctcggcgc gtctgcgtct ggctggctgg cataaatatc tcactcgcaa
     6841 tcaaattcag ccgatagcgg aacgggaagg cgactggagt gccatgtccg gttttcaaca
     6901 aaccatgcaa atgctgaatg agggcatcgt tcccactgcg atgctggttg ccaacgatca
     6961 gatggcgctg ggcgcaatgc gcgccattac cgagtccggg ctgcgcgttg gtgcggatat
     7021 ctcggtagtg ggatacgacg ataccgaaga cagctcatgt tatatcccgc cgttaaccac
     7081 catcaaacag gattttcgcc tgctggggca aaccagcgtg gaccgcttgc tgcaactctc
     7141 tcagggccag gcggtgaagg gcaatcagct gttgcccgtc tcactggtga aaagaaaaac
     7201 caccctggcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
     7261 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtaa
     7321 gttagctcac tcattaggca caattctcat gtttgacagc ttatcatcga ctgcacggtg
     7381 caccaatgct tctggcgtca ggcagccatc ggaagctgtg gtatggctgt gcaggtcgta
     7441 aatcactgca taattcgtgt cgctcaaggc gcactcccgt tctggataat gttttttgcg
     7501 ccgacatcat aacggttctg gcaaatattc tgaaatgagc tg
//
