LOCUS Exported 5490 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expresses human AR aa 1-173 in pGEX-5x-1 bacterial expression vector with GST. ACCESSION . VERSION . KEYWORDS GST-AR-1-173 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5490) AUTHORS He B, Kemppainen JA, Wilson EM TITLE FXXLF and WXXLF sequences mediate the NH2-terminal interaction with the ligand binding domain of the androgen receptor. JOURNAL J Biol Chem. 2000 Jul 28;275(30):22986-94. PUBMED 10816582 REFERENCE 2 (bases 1 to 5490) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5490 /organism="synthetic DNA construct" /mol_type="other DNA" rep_origin 27..615 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 516..535 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 769..786 /label=L4440 /note="L4440 vector, forward primer" promoter 859..936 /gene="lacI" /label=lacI promoter CDS 937..2019 /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(956..975) /label=LacI-R /note="LacI, reverse primer" promoter 2068..2098 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 2106..2122 /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 2130..2146 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 2130..2146 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" CDS 2142..2669 /codon_start=1 /gene="lacZ fragment" /product="LacZ-alpha fragment of beta-galactosidase" /label=lacZ-alpha /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR QPSEAVVWLCRS" primer_bind complement(2162..2179) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(2162..2178) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2171..2193) /label=M13/pUC Forward /note="In lacZ gene" promoter 2771..2799 /label=tac promoter /note="strong E. coli promoter; hybrid between the trp and lac UV5 promoters" protein_bind 2807..2823 /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)." CDS 2846..3499 /codon_start=1 /product="glutathione S-transferase from Schistosoma japonicum" /label=GST /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK" primer_bind 3457..3479 /label=pGEX 5' /note="pGEX vectors, Glutathione-S-transferase, forward primer" CDS 3509..3520 /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" primer_bind complement(4128..4150) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(4295..4313) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 4381..4485 /gene="bla" /label=AmpR promoter CDS 4486..5346 /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(4704..4723) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" ORIGIN 1 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc 61 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca 121 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta 181 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct 241 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg 301 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc 361 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta 421 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg 481 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt 541 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg 601 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg 661 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc 721 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg 781 agcgaggaag cggaagagcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt 841 tcacaccgca taaattccga caccatcgaa tggcgcaaaa cctttcgcgg tatggcatga 901 tagcgcccgg aagagagtca attcagggtg gtgaatgtga aaccagtaac gttatacgat 961 gtcgcagagt atgccggtgt ctcttatcag accgtttccc gcgtggtgaa ccaggccagc 1021 cacgtttctg cgaaaacgcg ggaaaaagtg gaagcggcga tggcggagct gaattacatt 1081 cccaaccgcg tggcacaaca actggcgggc aaacagtcgt tgctgattgg cgttgccacc 1141 tccagtctgg ccctgcacgc gccgtcgcaa attgtcgcgg cgattaaatc tcgcgccgat 1201 caactgggtg ccagcgtggt ggtgtcgatg gtagaacgaa gcggcgtcga agcctgtaaa 1261 gcggcggtgc acaatcttct cgcgcaacgc gtcagtgggc tgatcattaa ctatccgctg 1321 gatgaccagg atgccattgc tgtggaagct gcctgcacta atgttccggc gttatttctt 1381 gatgtctctg accagacacc catcaacagt attattttct cccatgaaga cggtacgcga 1441 ctgggcgtgg agcatctggt cgcattgggt caccagcaaa tcgcgctgtt agcgggccca 1501 ttaagttctg tctcggcgcg tctgcgtctg gctggctggc ataaatatct cactcgcaat 1561 caaattcagc cgatagcgga acgggaaggc gactggagtg ccatgtccgg ttttcaacaa 1621 accatgcaaa tgctgaatga gggcatcgtt cccactgcga tgctggttgc caacgatcag 1681 atggcgctgg gcgcaatgcg cgccattacc gagtccgggc tgcgcgttgg tgcggatatc 1741 tcggtagtgg gatacgacga taccgaagac agctcatgtt atatcccgcc gttaaccacc 1801 atcaaacagg attttcgcct gctggggcaa accagcgtgg accgcttgct gcaactctct 1861 cagggccagg cggtgaaggg caatcagctg ttgcccgtct cactggtgaa aagaaaaacc 1921 accctggcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag 1981 ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtaag 2041 ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg 2101 tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg attacggatt 2161 cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc 2221 gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc 2281 gcccttccca acagttgcgc agcctgaatg gcgaatggcg ctttgcctgg tttccggcac 2341 cagaagcggt gccggaaagc tggctggagt gcgatcttcc tgaggccgat actgtcgtcg 2401 tcccctcaaa ctggcagatg cacggttacg atgcgcccat ctacaccaac gtgacctatc 2461 ccattacggt caatccgccg tttgttccca cggagaatcc gacgggttgt tactcgctca 2521 catttaatgt tgatgaaagc tggctacagg aaggccagac gcgaattatt tttgatggcg 2581 ttggaattag cttatcgact gcacggtgca ccaatgcttc tggcgtcagg cagccatcgg 2641 aagctgtggt atggctgtgc aggtcgtaaa tcactgcata attcgtgtcg ctcaaggcgc 2701 actcccgttc tggataatgt tttttgcgcc gacatcataa cggttctggc aaatattctg 2761 aaatgagctg ttgacaatta atcatcggct cgtataatgt gtggaattgt gagcggataa 2821 caatttcaca caggaaacag tattcatgtc ccctatacta ggttattgga aaattaaggg 2881 ccttgtgcaa cccactcgac ttcttttgga atatcttgaa gaaaaatatg aagagcattt 2941 gtatgagcgc gatgaaggtg ataaatggcg aaacaaaaag tttgaattgg gtttggagtt 3001 tcccaatctt ccttattata ttgatggtga tgttaaatta acacagtcta tggccatcat 3061 acgttatata gctgacaagc acaacatgtt gggtggttgt ccaaaagagc gtgcagagat 3121 ttcaatgctt gaaggagcgg ttttggatat tagatacggt gtttcgagaa ttgcatatag 3181 taaagacttt gaaactctca aagttgattt tcttagcaag ctacctgaaa tgctgaaaat 3241 gttcgaagat cgtttatgtc ataaaacata tttaaatggt gatcatgtaa cccatcctga 3301 cttcatgttg tatgacgctc ttgatgttgt tttatacatg gacccaatgt gcctggatgc 3361 gttcccaaaa ttagtttgtt ttaaaaaacg tattgaagct atcccacaaa ttgataagta 3421 cttgaaatcc agcaagtata tagcatggcc tttgcagggc tggcaagcca cgtttggtgg 3481 tggcgaccat cctccaaaat cggatctgat cgaaggtcgt gggatccgtc gacatatgga 3541 agtgcagtta gggctgggaa gggtctaccc tcggccgccg tccaagacct accgaggagc 3601 tttccagaat ctgttccaga gcgtgcgcga agtgatccag aacccgggcc ccaggcaccc 3661 agaggccgcg agcgcagcac ctcccggcgc cagtttgctg ctgctgcagc agcagcagca 3721 gcagcagcag cagcagcagc agcagcagca gcagcaacag cagcagcagc agcagcagca 3781 agagactagc cccaggcagc agcagcagca gcagggtgag gatggttctc cccaagccca 3841 tcgtagaggc cccacaggct acctggtcct ggatgaggaa cagcaacctt cacagccgca 3901 gtcggccctg gagtgccacc ccgagagagg ttgcgtccca gagcctggag ccgccgtggc 3961 cgccagcaag gggctgccgc agcagctgcc agcacctccg gacgaggatg actcagctgc 4021 cccatccacg ttgtccctgc tgggccccac tttccccggc ttatcgagcg gccgcatcgt 4081 gactgactga cgatctgcct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg 4141 cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt 4201 cagggcgcgt cagcgggtgt tggcgggtgt cggggcgcag ccatgaccca gtcacgtagc 4261 gatagcggag tgtataattc ttgaagacga aagggcctcg tgatacgcct atttttatag 4321 gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg 4381 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga 4441 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat 4501 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca 4561 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc 4621 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca 4681 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtgt tgacgccggg 4741 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca 4801 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata 4861 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag 4921 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg 4981 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgc agcaatggca 5041 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta 5101 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct 5161 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca 5221 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag 5281 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat 5341 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt 5401 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa 5461 cgtgagtttt cgttccactg agcgtcagac //