LOCUS       sequence-199730-        6114 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human AR-174-566 in pGEX-3x bacterial expression vector as
            fusion protein with GST.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6114)
  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 6114)
  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: "J Biol 
            Chem."; date: "2000-07-28"; volume: "275(30)"; pages: "22986-94"
COMMENT     Sequence Label: GST-AR-174-566
FEATURES             Location/Qualifiers
     source          1..6114
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      21..609
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     510..529
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     763..780
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        853..930
                     /gene="lacI"
                     /label=lacI promoter
     CDS             931..2013
                     /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(950..969)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(2062..2067,2068..2085,2086..2092)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2100..2116
                     /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     2124..2140
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2124..2140
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2136..2663
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(2156..2173)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2156..2172)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2165..2187)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(2765..2770,2771..2786,2787..2793)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    2801..2817
                     /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             2840..3493
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     3451..3473
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             3503..3514
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4750..4772)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4913..4931)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4999..5103
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5104..5172,5173..5964)
                     /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(5322..5341)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
       61 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
      121 tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag
      181 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
      241 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
      301 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
      361 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
      421 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
      481 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
      541 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
      601 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
      661 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
      721 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
      781 gaagcggaag agcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac
      841 cgcataaatt ccgacaccat cgaatggcgc aaaacctttc gcggtatggc atgatagcgc
      901 ccggaagaga gtcaattcag ggtggtgaat gtgaaaccag taacgttata cgatgtcgca
      961 gagtatgccg gtgtctctta tcagaccgtt tcccgcgtgg tgaaccaggc cagccacgtt
     1021 tctgcgaaaa cgcgggaaaa agtggaagcg gcgatggcgg agctgaatta cattcccaac
     1081 cgcgtggcac aacaactggc gggcaaacag tcgttgctga ttggcgttgc cacctccagt
     1141 ctggccctgc acgcgccgtc gcaaattgtc gcggcgatta aatctcgcgc cgatcaactg
     1201 ggtgccagcg tggtggtgtc gatggtagaa cgaagcggcg tcgaagcctg taaagcggcg
     1261 gtgcacaatc ttctcgcgca acgcgtcagt gggctgatca ttaactatcc gctggatgac
     1321 caggatgcca ttgctgtgga agctgcctgc actaatgttc cggcgttatt tcttgatgtc
     1381 tctgaccaga cacccatcaa cagtattatt ttctcccatg aagacggtac gcgactgggc
     1441 gtggagcatc tggtcgcatt gggtcaccag caaatcgcgc tgttagcggg cccattaagt
     1501 tctgtctcgg cgcgtctgcg tctggctggc tggcataaat atctcactcg caatcaaatt
     1561 cagccgatag cggaacggga aggcgactgg agtgccatgt ccggttttca acaaaccatg
     1621 caaatgctga atgagggcat cgttcccact gcgatgctgg ttgccaacga tcagatggcg
     1681 ctgggcgcaa tgcgcgccat taccgagtcc gggctgcgcg ttggtgcgga tatctcggta
     1741 gtgggatacg acgataccga agacagctca tgttatatcc cgccgttaac caccatcaaa
     1801 caggattttc gcctgctggg gcaaaccagc gtggaccgct tgctgcaact ctctcagggc
     1861 caggcggtga agggcaatca gctgttgccc gtctcactgg tgaaaagaaa aaccaccctg
     1921 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
     1981 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg taagttagct
     2041 cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat
     2101 tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg gattcactgg
     2161 ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg
     2221 cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt
     2281 cccaacagtt gcgcagcctg aatggcgaat ggcgctttgc ctggtttccg gcaccagaag
     2341 cggtgccgga aagctggctg gagtgcgatc ttcctgaggc cgatactgtc gtcgtcccct
     2401 caaactggca gatgcacggt tacgatgcgc ccatctacac caacgtgacc tatcccatta
     2461 cggtcaatcc gccgtttgtt cccacggaga atccgacggg ttgttactcg ctcacattta
     2521 atgttgatga aagctggcta caggaaggcc agacgcgaat tatttttgat ggcgttggaa
     2581 ttagcttatc gactgcacgg tgcaccaatg cttctggcgt caggcagcca tcggaagctg
     2641 tggtatggct gtgcaggtcg taaatcactg cataattcgt gtcgctcaag gcgcactccc
     2701 gttctggata atgttttttg cgccgacatc ataacggttc tggcaaatat tctgaaatga
     2761 gctgttgaca attaatcatc ggctcgtata atgtgtggaa ttgtgagcgg ataacaattt
     2821 cacacaggaa acagtattca tgtcccctat actaggttat tggaaaatta agggccttgt
     2881 gcaacccact cgacttcttt tggaatatct tgaagaaaaa tatgaagagc atttgtatga
     2941 gcgcgatgaa ggtgataaat ggcgaaacaa aaagtttgaa ttgggtttgg agtttcccaa
     3001 tcttccttat tatattgatg gtgatgttaa attaacacag tctatggcca tcatacgtta
     3061 tatagctgac aagcacaaca tgttgggtgg ttgtccaaaa gagcgtgcag agatttcaat
     3121 gcttgaagga gcggttttgg atattagata cggtgtttcg agaattgcat atagtaaaga
     3181 ctttgaaact ctcaaagttg attttcttag caagctacct gaaatgctga aaatgttcga
     3241 agatcgttta tgtcataaaa catatttaaa tggtgatcat gtaacccatc ctgacttcat
     3301 gttgtatgac gctcttgatg ttgttttata catggaccca atgtgcctgg atgcgttccc
     3361 aaaattagtt tgttttaaaa aacgtattga agctatccca caaattgata agtacttgaa
     3421 atccagcaag tatatagcat ggcctttgca gggctggcaa gccacgtttg gtggtggcga
     3481 ccatcctcca aaatcggatc tgatcgaagg tcgtgggatc ttaagcagct gctccgctga
     3541 ccttaaagac atcctgagcg aggccagcac catgcaactc cttcagcaac agcagcagga
     3601 agcagtatcc gaaggcagca gcagcgggag agcgagggag gcctcggggg ctcccacttc
     3661 ctccaaggac aattacttag ggggcacttc gaccatttct gacaacgcca aggagttgtg
     3721 taaggcagtg tcggtgtcca tgggcctggg tgtggaggcg ttggagcatc tgagtccagg
     3781 ggaacagctt cggggggatt gcatgtacgc cccacttttg ggagttccac ccgctgtgcg
     3841 tcccactcct tgtgccccat tggccgaatg caaaggttct ctgctagacg acagcgcagg
     3901 caagagcact gaagatactg ctgagtattc ccctttcaag ggaggttaca ccaaagggct
     3961 agaaggcgag agcctaggct gctctggcag cgctgcagca gggagctccg ggacacttga
     4021 actgccgtct accctgtctc tctacaagtc cggagcactg gacgaggcag ctgcgtacca
     4081 gagtcgcgac tactacaact ttccactggc tctggccgga ccgccgcccc ctccgccgcc
     4141 tccccatccc cacgctcgca tcaagctgga gaacccgctg gactacggca gcgcctgggc
     4201 ggctgcggcg gcgcagtgcc gctatgggga cctggcgagc ctgcatggcg cgggtgcagc
     4261 gggacccggt tctgggtcac cctcagccgc cgcttcctca tcctggcaca ctctcttcac
     4321 agccgaagaa ggccagttgt atggaccgtg tggtggtggt gggggtggtg gcggcggcgg
     4381 cggcggcggc ggcggcggcg gcggcggcgg cgaggcggga gctgtagccc cctacggcta
     4441 cactcggccc cctcaggggc tggcgggcca ggaaagcgac ttcaccgcac ctgatgtgtg
     4501 gtaccctggc ggcatggtga gcagagtgcc ctatcccagt cccacttgtg tcaaaagcga
     4561 aatgggcccc tggatggata gctactccgg accttacggg gacatgcgtt tggagactgc
     4621 cagggaccat gttttgccca ttgactatta ctttccaccc cagaagacct gcctgatctg
     4681 tggagatgaa gctattcatc gtgactgact gacgatctgc ctcgcgcgtt tcggtgatga
     4741 cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga
     4801 tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggcgc
     4861 agccatgacc cagtcacgta gcgatagcgg agtaattctt gaagacgaaa gggcctcgtg
     4921 atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc
     4981 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     5041 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     5101 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     5161 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     5221 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     5281 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     5341 tcccgtgttg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     5401 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     5461 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     5521 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     5581 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     5641 atgcctgcag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     5701 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     5761 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     5821 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     5881 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     5941 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     6001 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     6061 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgta
//
