LOCUS Exported 7446 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Protein expression and purification of RXRA_Hormone-recep. ACCESSION . VERSION . KEYWORDS pHH0103_RXRA_Hormone-recep SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7446) TITLE Sidhu Lab peptide-recognition modules (PRMs) REFERENCE 2 (bases 1 to 7446) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7446 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 1..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 1641..1688 /label=fd terminator /note="central terminator from bacteriophage fd (Otsuka and Kunisawa, 1982)" rep_origin complement(3281..3736) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3418..3439) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 3630..3649 /label=F1ori-R /note="F1 origin, reverse primer" primer_bind complement(3778..3800) /label=pGEX 3' /note="pGEX vectors, reverse primer" misc_feature 3886..4026 /label=bom /note="basis of mobility region from pBR322" primer_bind 3941..3960 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind complement(4041..4058) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4212..4799) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(4970..5830) /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 5593..5612 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(5831..5935) /gene="bla" /label=AmpR promoter primer_bind 6003..6021 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 6044..6121 /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 6122..7204 /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(6141..6160) /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 cggccaccag ctctgcgaac gaagatatgc cggtggaacg tattctggaa gcagaactgg 961 ctgttgaacc gaaaaccgaa acgtatgtcg aagccaacat gggcctgaat ccgagttccc 1021 cgaacgatcc ggtcaccaat atttgccagg cggccgacaa acaactgttc accctggtgg 1081 aatgggcaaa gcgcatcccg catttttcag aactgccgct ggatgaccag gttattctgc 1141 tgcgtgcagg ttggaacgaa ctgctgatcg cttcattctc gcatcgctcg attgcggtca 1201 aagatggcat cctgctggcc acgggtctgc atgtgcaccg taatagcgcg cactctgccg 1261 gcgttggtgc aattttcgac cgtgtgctga ccgaactggt ttcaaaaatg cgcgatatgc 1321 aaatggacaa gacggaactg ggctgtctgc gcgctatcgt tctgtttaac ccggatagta 1381 aaggtctgtc caatccggca gaagtcgaag ctctgcgtga aaaggtgtac gcgagcctgg 1441 aagcctattg caaacacaag tacccggaac agccgggccg cttcgcaaaa ctgctgctgc 1501 gtctgccggc tctgcgctct atcggcctga aatgtctgga acacctgttt ttctttaagc 1561 tgattggtga taccccgatc gacacgtttc tgatggaaat gctggaagcc ccgcaccaaa 1621 tgaccgcggc cgcataataa accgatacaa ttaaaggctc cttttggagc cttttttttg 1681 gagattttca acgtgaaaaa attattattc gcaattcctt tagttgttcc tttctattct 1741 cactccgctg aaactgttga aagttgttta gcaaaacccc atacagaaaa ttcatttact 1801 aacgtctgga aagacgacaa aactttagat cgttacgcta actatgaggg ctgtctgtgg 1861 aatgctacag gcgttgtagt ttgtactggt gacgaaactc agtgtctagc tagagtggcg 1921 gtggctctgg ttccggtgat tttgattatg aaaagatggc aaacgctaat aagggggcta 1981 tgaccgaaaa tgccgatgaa aacgcgctac agtctgacgc taaaggcaaa cttgattctg 2041 tcgctactga ttacggtgct gctatcgatg gtttcattgg tgacgtttcc ggccttgcta 2101 atggtaatgg tgctactggt gattttgctg gctctaattc ccaaatggct caagtcggtg 2161 acggtgataa ttcaccttta atgaataatt tccgtcaata tttaccttcc ctccctcaat 2221 cggttgaatg tcgccctttt gtctttggcg ctggtaaacc atatgaattt tctattgatt 2281 gtgacaaaat aaacttattc cgtggtgtct ttgcgtttct tttatatgtt gccaccttta 2341 tgtatgtatt ttctacgttt gctaacatac tgcgtaataa ggagtcttaa tcatgccagt 2401 tcttttggct agcgccgccc tataccttgt ctgcctcccc gcgttgcgtc gcggtgcatg 2461 gagccgggcc acctcgacct gaatggaagc cggcggcacc tcgctaacgg attcaccact 2521 ccaagaattg gagccaatca attcttgcgg agaactgtga atgcgcaaac caacccttgg 2581 cagaacatat ccatcgcgtc cgccatctcc agcagccgca cgcggcgcat ctcgggcagc 2641 gttgggtcct ggccacgggt gcgcatgatc gtgctcctgt cgttgaggac ccggctaggc 2701 tggcggggtt gccttactgg ttagcagaat gaatcaccga tacgcgagcg aacgtgaagc 2761 gactgctgct gcaaaacgtc tgcgacctga gcaacaacat gaatggtctt cggtttccgt 2821 gtttcgtaaa gtctggaaac gcggaagtca gcgccctgca ccattatgtt ccggatctgc 2881 atcgcaggat gctgctggct accctgtgga acacctacat ctgtattaac gaagcgctgg 2941 cattgaccct gagtgatttt tctctggtcc cgccgcatcc ataccgccag ttgtttaccc 3001 tcacaacgtt ccagtaaccg ggcatgttca tcatcagtaa cccgtatcgt gagcatcctc 3061 tctcgtttca tcggtatcat tacccccatg aacagaaatc ccccttacac ggaggcatca 3121 gtgaccaaac aggaaaaaac cgcccttaac atggcccgct ttatcagaag ccagacatta 3181 acgcttctgg agaaactcaa cgagctggac gcggatgaac aggcagacat ctgtgaatcg 3241 cttcacgacc acgctgatga gctttaccgc aggatccgcg aaattgtaag cgttaatatt 3301 ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa 3361 atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca 3421 gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc 3481 gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg 3541 aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg 3601 ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg 3661 gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg 3721 ccgctacagg gcgcgtcgcg gatcctgcct cgcgcgtttc ggtgatgacg gtgaaaacct 3781 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag 3841 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggcgcag ccatgaccca 3901 gtcacgtagc gatagcggag tgtatactgg cttaactatg cggcatcaga gcagattgta 3961 ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc 4021 atcaggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg 4081 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac 4141 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg 4201 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca 4261 agtcagaggt ggcgaaaccc gacaggacta taaagatacc agggtttccc cctggaagct 4321 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc 4381 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg 4441 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct 4501 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag 4561 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga 4621 agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga 4681 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg 4741 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag 4801 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag 4861 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat 4921 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct 4981 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac 5041 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa 5101 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg 5161 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt 5221 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca 5281 ttgctgcagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt 5341 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct 5401 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg 5461 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg 5521 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg 5581 cgtcaacacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa 5641 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt 5701 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt 5761 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt 5821 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca 5881 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat 5941 ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata 6001 aaaataggcg tatcacgagg ccctttcgtc ttcaagaatt cccgacacca tcgaatggtg 6061 caaaaccttt cgcggtatgg catgatagcg cccggaagag agtcaattca gggtggtgaa 6121 tgtgaaacca gtaacgttat acgatgtcgc agagtatgcc ggtgtctctt atcagaccgt 6181 ttcccgcgtg gtgaaccagg ccagccacgt ttctgcgaaa acgcgggaaa aagtggaagc 6241 ggcgatggcg gagctgaatt acattcccaa ccgcgtggca caacaactgg cgggcaaaca 6301 gtcgttgctg attggcgttg ccacctccag tctggccctg cacgcgccgt cgcaaattgt 6361 cgcggcgatt aaatctcgcg ccgatcaact gggtgccagc gtggtggtgt cgatggtaga 6421 acgaagcggc gtcgaagcct gtaaagcggc ggtgcacaat cttctcgcgc aacgcgtcag 6481 tgggctgatc attaactatc cgctggatga ccaggatgcc attgctgtgg aagctgcctg 6541 cactaatgtt ccggcgttat ttcttgatgt ctctgaccag acacccatca acagtattat 6601 tttctcccat gaagacggta cgcgactggg cgtggagcat ctggtcgcat tgggtcacca 6661 gcaaatcgcg ctgttagcgg gcccattaag ttctgtctcg gcgcgtctgc gtctggctgg 6721 ctggcataaa tatctcactc gcaatcaaat tcagccgata gcggaacggg aaggcgactg 6781 gagtgccatg tccggttttc aacaaaccat gcaaatgctg aatgagggca tcgttcccac 6841 tgcgatgctg gttgccaacg atcagatggc gctgggcgca atgcgcgcca ttaccgagtc 6901 cgggctgcgc gttggtgcgg atatctcggt agtgggatac gacgataccg aagacagctc 6961 atgttatatc ccgccgttaa ccaccatcaa acaggatttt cgcctgctgg ggcaaaccag 7021 cgtggaccgc ttgctgcaac tctctcaggg ccaggcggtg aagggcaatc agctgttgcc 7081 cgtctcactg gtgaaaagaa aaaccaccct ggcgcccaat acgcaaaccg cctctccccg 7141 cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca 7201 gtgagcgcaa cgcaattaat gtaagttagc tcactcatta ggcacaattc tcatgtttga 7261 cagcttatca tcgactgcac ggtgcaccaa tgcttctggc gtcaggcagc catcggaagc 7321 tgtggtatgg ctgtgcaggt cgtaaatcac tgcataattc gtgtcgctca aggcgcactc 7381 ccgttctgga taatgttttt tgcgccgaca tcataacggt tctggcaaat attctgaaat 7441 gagctg //