LOCUS       sequence-217270-        7503 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Protein expression and purification of RORB_Hormone-recep.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7503)
  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 7503)
  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_RORB_Hormone-recep
FEATURES             Location/Qualifiers
     source          1..7503
                     /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      1698..1745
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      complement(3338..3793)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3475..3496)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3687..3706
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     complement(3835..3857)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3943..4083
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3998..4017
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4098..4115)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4269..4856)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(5027..5818,5819..5887))
                     /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     5650..5669
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5888..5992)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6060..6078
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6101..6178
                     /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             6179..7261
                     /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(6198..6217)
                     /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 cggccggtca gctggcaccg ggtattacca tgagcgaaat tgatcgtatt gcccagaaca
      961 tcattaaaag ccatctggaa acctgtcagt acaccatgga agaactgcac cagctggcct
     1021 ggcagaccca tacctatgaa gaaattaaag cctatcagag caaaagccgt gaagcactgt
     1081 ggcagcagtg tgcaattcag attacccatg ccattcagta tgttgtggaa tttgcaaaac
     1141 gcatcaccgg ttttatggaa ctgtgtcaga atgatcagat tctgctgctg aaaagcggtt
     1201 gtctggaagt tgttctggtt cgtatgtgtc gtgcatttaa tccgctgaat aacaccgttc
     1261 tgtttgaagg taaatatggt ggcatgcaga tgtttaaagc actgggtagt gatgatctgg
     1321 tgaatgaagc atttgatttc gccaaaaatc tgtgtagcct gcagctgaca gaagaagaaa
     1381 tcgcactgtt tagcagcgca gttctgatta gtccggatcg tgcatggctg ctggaaccgc
     1441 gtaaagttca gaaactgcaa gagaaaatct attttgccct gcagcatgtg atccagaaaa
     1501 atcatctgga tgatgaaacc ctggcaaaac tgattgcaaa aattccgacc attaccgcag
     1561 tttgtaacct gcatggtgaa aaactgcagg tatttaaaca gagccatccg gatattgtga
     1621 ataccctgtt tccgcctctg tataaagaac tgtttaaccc ggattgtgcc accggttgta
     1681 aagcggccgc ataataaacc gatacaatta aaggctcctt ttggagcctt ttttttggag
     1741 attttcaacg tgaaaaaatt attattcgca attcctttag ttgttccttt ctattctcac
     1801 tccgctgaaa ctgttgaaag ttgtttagca aaaccccata cagaaaattc atttactaac
     1861 gtctggaaag acgacaaaac tttagatcgt tacgctaact atgagggctg tctgtggaat
     1921 gctacaggcg ttgtagtttg tactggtgac gaaactcagt gtctagctag agtggcggtg
     1981 gctctggttc cggtgatttt gattatgaaa agatggcaaa cgctaataag ggggctatga
     2041 ccgaaaatgc cgatgaaaac gcgctacagt ctgacgctaa aggcaaactt gattctgtcg
     2101 ctactgatta cggtgctgct atcgatggtt tcattggtga cgtttccggc cttgctaatg
     2161 gtaatggtgc tactggtgat tttgctggct ctaattccca aatggctcaa gtcggtgacg
     2221 gtgataattc acctttaatg aataatttcc gtcaatattt accttccctc cctcaatcgg
     2281 ttgaatgtcg cccttttgtc tttggcgctg gtaaaccata tgaattttct attgattgtg
     2341 acaaaataaa cttattccgt ggtgtctttg cgtttctttt atatgttgcc acctttatgt
     2401 atgtattttc tacgtttgct aacatactgc gtaataagga gtcttaatca tgccagttct
     2461 tttggctagc gccgccctat accttgtctg cctccccgcg ttgcgtcgcg gtgcatggag
     2521 ccgggccacc tcgacctgaa tggaagccgg cggcacctcg ctaacggatt caccactcca
     2581 agaattggag ccaatcaatt cttgcggaga actgtgaatg cgcaaaccaa cccttggcag
     2641 aacatatcca tcgcgtccgc catctccagc agccgcacgc ggcgcatctc gggcagcgtt
     2701 gggtcctggc cacgggtgcg catgatcgtg ctcctgtcgt tgaggacccg gctaggctgg
     2761 cggggttgcc ttactggtta gcagaatgaa tcaccgatac gcgagcgaac gtgaagcgac
     2821 tgctgctgca aaacgtctgc gacctgagca acaacatgaa tggtcttcgg tttccgtgtt
     2881 tcgtaaagtc tggaaacgcg gaagtcagcg ccctgcacca ttatgttccg gatctgcatc
     2941 gcaggatgct gctggctacc ctgtggaaca cctacatctg tattaacgaa gcgctggcat
     3001 tgaccctgag tgatttttct ctggtcccgc cgcatccata ccgccagttg tttaccctca
     3061 caacgttcca gtaaccgggc atgttcatca tcagtaaccc gtatcgtgag catcctctct
     3121 cgtttcatcg gtatcattac ccccatgaac agaaatcccc cttacacgga ggcatcagtg
     3181 accaaacagg aaaaaaccgc ccttaacatg gcccgcttta tcagaagcca gacattaacg
     3241 cttctggaga aactcaacga gctggacgcg gatgaacagg cagacatctg tgaatcgctt
     3301 cacgaccacg ctgatgagct ttaccgcagg atccgcgaaa ttgtaagcgt taatattttg
     3361 ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc
     3421 ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt
     3481 tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc
     3541 tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg
     3601 tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga
     3661 aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg
     3721 ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg
     3781 ctacagggcg cgtcgcggat cctgcctcgc gcgtttcggt gatgacggtg aaaacctctg
     3841 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca
     3901 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggcgcagcca tgacccagtc
     3961 acgtagcgat agcggagtgt atactggctt aactatgcgg catcagagca gattgtactg
     4021 agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc
     4081 aggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     4141 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     4201 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     4261 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     4321 cagaggtggc gaaacccgac aggactataa agataccagg gtttccccct ggaagctccc
     4381 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     4441 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     4501 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     4561 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     4621 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     4681 ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc
     4741 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     4801 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     4861 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     4921 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     4981 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     5041 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     5101 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     5161 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     5221 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     5281 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     5341 ctgcaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     5401 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     5461 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     5521 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     5581 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     5641 caacacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     5701 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     5761 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     5821 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     5881 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     5941 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     6001 cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa
     6061 ataggcgtat cacgaggccc tttcgtcttc aagaattccc gacaccatcg aatggtgcaa
     6121 aacctttcgc ggtatggcat gatagcgccc ggaagagagt caattcaggg tggtgaatgt
     6181 gaaaccagta acgttatacg atgtcgcaga gtatgccggt gtctcttatc agaccgtttc
     6241 ccgcgtggtg aaccaggcca gccacgtttc tgcgaaaacg cgggaaaaag tggaagcggc
     6301 gatggcggag ctgaattaca ttcccaaccg cgtggcacaa caactggcgg gcaaacagtc
     6361 gttgctgatt ggcgttgcca cctccagtct ggccctgcac gcgccgtcgc aaattgtcgc
     6421 ggcgattaaa tctcgcgccg atcaactggg tgccagcgtg gtggtgtcga tggtagaacg
     6481 aagcggcgtc gaagcctgta aagcggcggt gcacaatctt ctcgcgcaac gcgtcagtgg
     6541 gctgatcatt aactatccgc tggatgacca ggatgccatt gctgtggaag ctgcctgcac
     6601 taatgttccg gcgttatttc ttgatgtctc tgaccagaca cccatcaaca gtattatttt
     6661 ctcccatgaa gacggtacgc gactgggcgt ggagcatctg gtcgcattgg gtcaccagca
     6721 aatcgcgctg ttagcgggcc cattaagttc tgtctcggcg cgtctgcgtc tggctggctg
     6781 gcataaatat ctcactcgca atcaaattca gccgatagcg gaacgggaag gcgactggag
     6841 tgccatgtcc ggttttcaac aaaccatgca aatgctgaat gagggcatcg ttcccactgc
     6901 gatgctggtt gccaacgatc agatggcgct gggcgcaatg cgcgccatta ccgagtccgg
     6961 gctgcgcgtt ggtgcggata tctcggtagt gggatacgac gataccgaag acagctcatg
     7021 ttatatcccg ccgttaacca ccatcaaaca ggattttcgc ctgctggggc aaaccagcgt
     7081 ggaccgcttg ctgcaactct ctcagggcca ggcggtgaag ggcaatcagc tgttgcccgt
     7141 ctcactggtg aaaagaaaaa ccaccctggc gcccaatacg caaaccgcct ctccccgcgc
     7201 gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg
     7261 agcgcaacgc aattaatgta agttagctca ctcattaggc acaattctca tgtttgacag
     7321 cttatcatcg actgcacggt gcaccaatgc ttctggcgtc aggcagccat cggaagctgt
     7381 ggtatggctg tgcaggtcgt aaatcactgc ataattcgtg tcgctcaagg cgcactcccg
     7441 ttctggataa tgttttttgc gccgacatca taacggttct ggcaaatatt ctgaaatgag
     7501 ctg
//
