LOCUS       sequence-217277-        7485 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Protein expression and purification of VDR_Hormone-recep.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7485)
  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 7485)
  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_VDR_Hormone-recep
FEATURES             Location/Qualifiers
     source          1..7485
                     /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      1680..1727
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      complement(3320..3775)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3457..3478)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3669..3688
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     complement(3817..3839)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3925..4065
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3980..3999
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4080..4097)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4251..4838)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(5009..5800,5801..5869))
                     /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     5632..5651
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5870..5974)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6042..6060
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6083..6160
                     /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             6161..7243
                     /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(6180..6199)
                     /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 cggccttacg tccgaaactg agcgaagaac agcagcgtat tattgcaatt ctgctggatg
      961 cacatcacaa aacctatgat ccgacctata gcgatttttg ccagtttcgt ccgcctgttc
     1021 gtgttaatga tggtggtggt agcgttaccc tggaactgag ccagctgagc atgctgccgc
     1081 atctggcaga tctggttagc tatagcattc agaaagttat cggcttcgca aaaatgattc
     1141 cgggttttcg tgatctgacc agcgaagatc agattgttct gctgaaaagc agcgccattg
     1201 aagttattat gctgcgtagc aatgaaagct tcaccatgga tgatatgagc tggacctgtg
     1261 gtaatcagga ttacaaatat cgtgttagcg acgttaccaa agcaggtcat agcctggaat
     1321 taattgaacc gctgatcaaa tttcaggtgg gcctgaaaaa actgaatctg catgaagaag
     1381 aacacgtact gctgatggca atttgtattg ttagtccgga tcgtccgggt gttcaggatg
     1441 cagcactgat tgaagcaatt caggatcgtc tgagcaatac cctgcagacc tatattcgtt
     1501 gtcgtcatcc gcctccgggt agccatctgc tgtatgccaa aatgatccaa aaactggccg
     1561 atctgcgttc actgaatgag gaacatagca aacagtatcg ttgtctgagc tttcagccgg
     1621 aatgtagcat gaaactgaca ccgctggttc tggaagtgtt tggtgcggcc gcataataaa
     1681 ccgatacaat taaaggctcc ttttggagcc ttttttttgg agattttcaa cgtgaaaaaa
     1741 ttattattcg caattccttt agttgttcct ttctattctc actccgctga aactgttgaa
     1801 agttgtttag caaaacccca tacagaaaat tcatttacta acgtctggaa agacgacaaa
     1861 actttagatc gttacgctaa ctatgagggc tgtctgtgga atgctacagg cgttgtagtt
     1921 tgtactggtg acgaaactca gtgtctagct agagtggcgg tggctctggt tccggtgatt
     1981 ttgattatga aaagatggca aacgctaata agggggctat gaccgaaaat gccgatgaaa
     2041 acgcgctaca gtctgacgct aaaggcaaac ttgattctgt cgctactgat tacggtgctg
     2101 ctatcgatgg tttcattggt gacgtttccg gccttgctaa tggtaatggt gctactggtg
     2161 attttgctgg ctctaattcc caaatggctc aagtcggtga cggtgataat tcacctttaa
     2221 tgaataattt ccgtcaatat ttaccttccc tccctcaatc ggttgaatgt cgcccttttg
     2281 tctttggcgc tggtaaacca tatgaatttt ctattgattg tgacaaaata aacttattcc
     2341 gtggtgtctt tgcgtttctt ttatatgttg ccacctttat gtatgtattt tctacgtttg
     2401 ctaacatact gcgtaataag gagtcttaat catgccagtt cttttggcta gcgccgccct
     2461 ataccttgtc tgcctccccg cgttgcgtcg cggtgcatgg agccgggcca cctcgacctg
     2521 aatggaagcc ggcggcacct cgctaacgga ttcaccactc caagaattgg agccaatcaa
     2581 ttcttgcgga gaactgtgaa tgcgcaaacc aacccttggc agaacatatc catcgcgtcc
     2641 gccatctcca gcagccgcac gcggcgcatc tcgggcagcg ttgggtcctg gccacgggtg
     2701 cgcatgatcg tgctcctgtc gttgaggacc cggctaggct ggcggggttg ccttactggt
     2761 tagcagaatg aatcaccgat acgcgagcga acgtgaagcg actgctgctg caaaacgtct
     2821 gcgacctgag caacaacatg aatggtcttc ggtttccgtg tttcgtaaag tctggaaacg
     2881 cggaagtcag cgccctgcac cattatgttc cggatctgca tcgcaggatg ctgctggcta
     2941 ccctgtggaa cacctacatc tgtattaacg aagcgctggc attgaccctg agtgattttt
     3001 ctctggtccc gccgcatcca taccgccagt tgtttaccct cacaacgttc cagtaaccgg
     3061 gcatgttcat catcagtaac ccgtatcgtg agcatcctct ctcgtttcat cggtatcatt
     3121 acccccatga acagaaatcc cccttacacg gaggcatcag tgaccaaaca ggaaaaaacc
     3181 gcccttaaca tggcccgctt tatcagaagc cagacattaa cgcttctgga gaaactcaac
     3241 gagctggacg cggatgaaca ggcagacatc tgtgaatcgc ttcacgacca cgctgatgag
     3301 ctttaccgca ggatccgcga aattgtaagc gttaatattt tgttaaaatt cgcgttaaat
     3361 ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa
     3421 tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta
     3481 ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca
     3541 ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat
     3601 cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg
     3661 agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc
     3721 acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcgcgg
     3781 atcctgcctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga
     3841 gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc
     3901 agcgggtgtt ggcgggtgtc ggggcgcagc catgacccag tcacgtagcg atagcggagt
     3961 gtatactggc ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatgcgg
     4021 tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgctc ttccgcttcc
     4081 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     4141 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     4201 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     4261 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     4321 acaggactat aaagatacca gggtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     4381 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     4441 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     4501 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     4561 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     4621 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     4681 acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     4741 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     4801 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     4861 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     4921 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     4981 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     5041 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     5101 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     5161 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     5221 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     5281 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctgcaggc atcgtggtgt
     5341 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     5401 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     5461 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     5521 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     5581 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaacacgg gataataccg
     5641 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     5701 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     5761 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     5821 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     5881 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     5941 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     6001 acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc
     6061 cctttcgtct tcaagaattc ccgacaccat cgaatggtgc aaaacctttc gcggtatggc
     6121 atgatagcgc ccggaagaga gtcaattcag ggtggtgaat gtgaaaccag taacgttata
     6181 cgatgtcgca gagtatgccg gtgtctctta tcagaccgtt tcccgcgtgg tgaaccaggc
     6241 cagccacgtt tctgcgaaaa cgcgggaaaa agtggaagcg gcgatggcgg agctgaatta
     6301 cattcccaac cgcgtggcac aacaactggc gggcaaacag tcgttgctga ttggcgttgc
     6361 cacctccagt ctggccctgc acgcgccgtc gcaaattgtc gcggcgatta aatctcgcgc
     6421 cgatcaactg ggtgccagcg tggtggtgtc gatggtagaa cgaagcggcg tcgaagcctg
     6481 taaagcggcg gtgcacaatc ttctcgcgca acgcgtcagt gggctgatca ttaactatcc
     6541 gctggatgac caggatgcca ttgctgtgga agctgcctgc actaatgttc cggcgttatt
     6601 tcttgatgtc tctgaccaga cacccatcaa cagtattatt ttctcccatg aagacggtac
     6661 gcgactgggc gtggagcatc tggtcgcatt gggtcaccag caaatcgcgc tgttagcggg
     6721 cccattaagt tctgtctcgg cgcgtctgcg tctggctggc tggcataaat atctcactcg
     6781 caatcaaatt cagccgatag cggaacggga aggcgactgg agtgccatgt ccggttttca
     6841 acaaaccatg caaatgctga atgagggcat cgttcccact gcgatgctgg ttgccaacga
     6901 tcagatggcg ctgggcgcaa tgcgcgccat taccgagtcc gggctgcgcg ttggtgcgga
     6961 tatctcggta gtgggatacg acgataccga agacagctca tgttatatcc cgccgttaac
     7021 caccatcaaa caggattttc gcctgctggg gcaaaccagc gtggaccgct tgctgcaact
     7081 ctctcagggc caggcggtga agggcaatca gctgttgccc gtctcactgg tgaaaagaaa
     7141 aaccaccctg gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat
     7201 gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg
     7261 taagttagct cactcattag gcacaattct catgtttgac agcttatcat cgactgcacg
     7321 gtgcaccaat gcttctggcg tcaggcagcc atcggaagct gtggtatggc tgtgcaggtc
     7381 gtaaatcact gcataattcg tgtcgctcaa ggcgcactcc cgttctggat aatgtttttt
     7441 gcgccgacat cataacggtt ctggcaaata ttctgaaatg agctg
//
