LOCUS Exported 5629 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expresses human AR-334-566 in pGEX-4T-1 bacterial expression vector as fusion protein with GST. ACCESSION . VERSION . KEYWORDS GST-AR-334-566 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5629) 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 5629) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5629 /organism="synthetic DNA construct" /mol_type="other DNA" rep_origin 18..606 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 507..526 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 760..777 /label=L4440 /note="L4440 vector, forward primer" promoter 850..927 /gene="lacI" /label=lacI promoter CDS 928..2010 /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(947..966) /label=LacI-R /note="LacI, reverse primer" promoter 2059..2089 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 2097..2113 /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 2121..2137 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 2121..2137 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" CDS 2133..2660 /codon_start=1 /gene="lacZ fragment" /product="LacZ-alpha fragment of beta-galactosidase" /label=lacZ-alpha /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR QPSEAVVWLCRS" primer_bind complement(2153..2170) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(2153..2169) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2162..2184) /label=M13/pUC Forward /note="In lacZ gene" promoter 2762..2790 /label=tac promoter /note="strong E. coli promoter; hybrid between the trp and lac UV5 promoters" protein_bind 2798..2814 /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 2837..3490 /codon_start=1 /product="glutathione S-transferase from Schistosoma japonicum" /label=GST /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK" primer_bind 3448..3470 /label=pGEX 5' /note="pGEX vectors, Glutathione-S-transferase, forward primer" CDS 3497..3514 /codon_start=1 /product="thrombin recognition and cleavage site" /label=thrombin site /translation="LVPRGS" primer_bind complement(4258..4280) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(4425..4443) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 4511..4615 /gene="bla" /label=AmpR promoter CDS 4616..5476 /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(4834..4853) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" ORIGIN 1 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca 61 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt 121 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg 181 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc 241 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga 301 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc 361 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc 421 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca 481 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg 541 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta 601 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct 661 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag 721 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa 781 gcggaagagc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc 841 ataaattccg acaccatcga atggcgcaaa acctttcgcg gtatggcatg atagcgcccg 901 gaagagagtc aattcagggt ggtgaatgtg aaaccagtaa cgttatacga tgtcgcagag 961 tatgccggtg tctcttatca gaccgtttcc cgcgtggtga accaggccag ccacgtttct 1021 gcgaaaacgc gggaaaaagt ggaagcggcg atggcggagc tgaattacat tcccaaccgc 1081 gtggcacaac aactggcggg caaacagtcg ttgctgattg gcgttgccac ctccagtctg 1141 gccctgcacg cgccgtcgca aattgtcgcg gcgattaaat ctcgcgccga tcaactgggt 1201 gccagcgtgg tggtgtcgat ggtagaacga agcggcgtcg aagcctgtaa agcggcggtg 1261 cacaatcttc tcgcgcaacg cgtcagtggg ctgatcatta actatccgct ggatgaccag 1321 gatgccattg ctgtggaagc tgcctgcact aatgttccgg cgttatttct tgatgtctct 1381 gaccagacac ccatcaacag tattattttc tcccatgaag acggtacgcg actgggcgtg 1441 gagcatctgg tcgcattggg tcaccagcaa atcgcgctgt tagcgggccc attaagttct 1501 gtctcggcgc gtctgcgtct ggctggctgg cataaatatc tcactcgcaa tcaaattcag 1561 ccgatagcgg aacgggaagg cgactggagt gccatgtccg gttttcaaca aaccatgcaa 1621 atgctgaatg agggcatcgt tcccactgcg atgctggttg ccaacgatca gatggcgctg 1681 ggcgcaatgc gcgccattac cgagtccggg ctgcgcgttg gtgcggatat ctcggtagtg 1741 ggatacgacg ataccgaaga cagctcatgt tatatcccgc cgttaaccac catcaaacag 1801 gattttcgcc tgctggggca aaccagcgtg gaccgcttgc tgcaactctc tcagggccag 1861 gcggtgaagg gcaatcagct gttgcccgtc tcactggtga aaagaaaaac caccctggcg 1921 cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga 1981 caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtaa gttagctcac 2041 tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt 2101 gagcggataa caatttcaca caggaaacag ctatgaccat gattacggat tcactggccg 2161 tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag 2221 cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc 2281 aacagttgcg cagcctgaat ggcgaatggc gctttgcctg gtttccggca ccagaagcgg 2341 tgccggaaag ctggctggag tgcgatcttc ctgaggccga tactgtcgtc gtcccctcaa 2401 actggcagat gcacggttac gatgcgccca tctacaccaa cgtgacctat cccattacgg 2461 tcaatccgcc gtttgttccc acggagaatc cgacgggttg ttactcgctc acatttaatg 2521 ttgatgaaag ctggctacag gaaggccaga cgcgaattat ttttgatggc gttggaatta 2581 gcttatcgac tgcacggtgc accaatgctt ctggcgtcag gcagccatcg gaagctgtgg 2641 tatggctgtg caggtcgtaa atcactgcat aattcgtgtc gctcaaggcg cactcccgtt 2701 ctggataatg ttttttgcgc cgacatcata acggttctgg caaatattct gaaatgagct 2761 gttgacaatt aatcatcggc tcgtataatg tgtggaattg tgagcggata acaatttcac 2821 acaggaaaca gtattcatgt cccctatact aggttattgg aaaattaagg gccttgtgca 2881 acccactcga cttcttttgg aatatcttga agaaaaatat gaagagcatt tgtatgagcg 2941 cgatgaaggt gataaatggc gaaacaaaaa gtttgaattg ggtttggagt ttcccaatct 3001 tccttattat attgatggtg atgttaaatt aacacagtct atggccatca tacgttatat 3061 agctgacaag cacaacatgt tgggtggttg tccaaaagag cgtgcagaga tttcaatgct 3121 tgaaggagcg gttttggata ttagatacgg tgtttcgaga attgcatata gtaaagactt 3181 tgaaactctc aaagttgatt ttcttagcaa gctacctgaa atgctgaaaa tgttcgaaga 3241 tcgtttatgt cataaaacat atttaaatgg tgatcatgta acccatcctg acttcatgtt 3301 gtatgacgct cttgatgttg ttttatacat ggacccaatg tgcctggatg cgttcccaaa 3361 attagtttgt tttaaaaaac gtattgaagc tatcccacaa attgataagt acttgaaatc 3421 cagcaagtat atagcatggc ctttgcaggg ctggcaagcc acgtttggtg gtggcgacca 3481 tcctccaaaa tcggatctgg ttccgcgtgg atccgggaca cttgaactgc cgtctaccct 3541 gtctctctac aagtccggag cactggacga ggcagctgcg taccagagtc gcgactacta 3601 caactttcca ctggctctgg ccggaccgcc gccccctccg ccgcctcccc atccccacgc 3661 tcgcatcaag ctggagaacc cgctggacta cggcagcgcc tgggcggctg cggcggcgca 3721 gtgccgctat ggggacctgg cgagcctgca tggcgcgggt gcagcgggac ccggttctgg 3781 gtcaccctca gccgccgctt cctcatcctg gcacactctc ttcacagccg aagaaggcca 3841 gttgtatgga ccgtgtggtg gtggtggggg tggtggcggc ggcggcggcg gcggcggcgg 3901 cggcggcggc ggcgaggcgg gagctgtagc cccctacggc tacactcggc cccctcaggg 3961 gctggcgggc caggaaagcg acttcaccgc acctgatgtg tggtaccctg gcggcatggt 4021 gagcagagtg ccctatccca gtcccacttg tgtcaaaagc gaaatgggcc cctggatgga 4081 tagctactcc ggaccttacg gggacatgcg tttggagact gccagggacc atgttttgcc 4141 cattgactat tactttccac cccagaagac ctgcctgatc tgtggagatg aactcgagcg 4201 gccgcatcgt gactgactga cgatctgcct cgcgcgtttc ggtgatgacg gtgaaaacct 4261 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag 4321 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggcgcag ccatgaccca 4381 gtcacgtagc gatagcggag tgtataattc ttgaagacga aagggcctcg tgatacgcct 4441 atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg 4501 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc 4561 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag 4621 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt 4681 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt 4741 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga 4801 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtgt 4861 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga 4921 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag 4981 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg 5041 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg 5101 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgc 5161 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg 5221 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc 5281 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg 5341 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac 5401 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact 5461 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa 5521 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa 5581 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaa //