LOCUS       Exported                6474 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  bacterial expression of human androgen receptor ligand bindng domain
            amino acid residues 663-919 with histidine tag.
ACCESSION   .
VERSION     .
KEYWORDS    pET-15b-hAR-663-919
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6474)
  AUTHORS   Askew EB, Gampe RT Jr, Stanley TB, Faggart JL, Wilson EM
  TITLE     Modulation of androgen receptor activation function 2 by 
            testosterone and dihydrotestosterone.
  JOURNAL   J Biol Chem. 2007 Aug 31;282(35):25801-16. doi: 
            10.1074/jbc.M703268200. Epub 2007 Jun 25.
  PUBMED    17591767
REFERENCE   2  (bases 1 to 6474)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6474
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    20..44
                     /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)."
     RBS             59..81
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             100..117
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             127..144
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(975..993)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      989..1036
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(1211..1239)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(1266..1284)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1352..1456
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1457..2317
                     /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(1675..1694)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2488..3076
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2977..2996
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3230..3247
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3262..3401
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3327..3346)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3487..3509
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3503..3694)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5006..6088)
                     /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     6050..6069
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6089..6166)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6372..6391
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atataccatg ggcagcagcc atcatcatca tcatcacagc
      121 agcggcctgg tgccgcgcgg cagccatatg cacattgaag gctatgaatg tcagcccatc
      181 tttctgaatg tcctggaagc cattgagcca ggtgtagtgt gtgctggaca cgacaacaac
      241 cagcccgact cctttgcagc cttgctctct agcctcaatg aactgggaga gagacagctt
      301 gtacacgtgg tcaagtgggc caaggccttg cctggcttcc gcaacttaca cgtggacgac
      361 cagatggctg tcattcagta ctcctggatg gggctcatgg tgtttgccat gggctggcga
      421 tccttcacca atgtcaactc caggatgctc tacttcgccc ctgatctggt tttcaatgag
      481 taccgcatgc acaagtcccg gatgtacagc cagtgtgtcc gaatgaggca cctctctcaa
      541 gagtttggat ggctccaaat caccccccag gaattcctgt gcatgaaagc actgctactc
      601 ttcagcatta ttccagtgga tgggctgaaa aatcaaaaat tctttgatga acttcgaatg
      661 aactacatca aggaactcga tcgtatcatt gcatgcaaaa gaaaaaatcc cacatcctgc
      721 tcaagacgct tctaccagct caccaagctc ctggactccg tgcagcctat tgcgagagag
      781 ctgcatcagt tcacttttga cctgctaatc aagtcacaca tggtgagcgt ggactttccg
      841 gaaatgatgg cagagatcat ctctgtgcaa gtgcccaaga tcctttctgg gaaagtcaag
      901 cccatctatt tccacaccca gtgaggatcc ggctgctaac aaagcccgaa aggaagctga
      961 gttggctgct gccaccgctg agcaataact agcataaccc cttggggcct ctaaacgggt
     1021 cttgaggggt tttttgctga aaggaggaac tatatccgga tatcccgcaa gaggcccggc
     1081 agtaccggca taaccaagcc tatgcctaca gcatccaggg tgacggtgcc gaggatgacg
     1141 atgagcgcat tgttagattt catacacggt gcctgactgc gttagcaatt taactgtgat
     1201 aaactaccgc attaaagctt atcgatgata agctgtcaaa catgagaatt cttgaagacg
     1261 aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta
     1321 gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     1381 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     1441 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     1501 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     1561 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     1621 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     1681 tggcgcggta ttatcccgtg ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     1741 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     1801 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     1861 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     1921 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     1981 gcgtgacacc acgatgcctg cagcaatggc aacaacgttg cgcaaactat taactggcga
     2041 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     2101 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     2161 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     2221 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     2281 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     2341 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     2401 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     2461 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     2521 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     2581 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct
     2641 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     2701 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     2761 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     2821 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     2881 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     2941 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     3001 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     3061 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     3121 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     3181 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
     3241 gagcgaggaa gcggaagagc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat
     3301 ttcacaccgc aatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag
     3361 tatacactcc gctatcgcta cgtgactggg tcatggctgc gccccgacac ccgccaacac
     3421 ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga
     3481 ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgaggc
     3541 agctgcggta aagctcatca gcgtggtcgt gaagcgattc acagatgtct gcctgttcat
     3601 ccgcgtccag ctcgttgagt ttctccagaa gcgttaatgt ctggcttctg ataaagcggg
     3661 ccatgttaag ggcggttttt tcctgtttgg tcactgatgc ctccgtgtaa gggggatttc
     3721 tgttcatggg ggtaatgata ccgatgaaac gagagaggat gctcacgata cgggttactg
     3781 atgatgaaca tgcccggtta ctggaacgtt gtgagggtaa acaactggcg gtatggatgc
     3841 ggcgggacca gagaaaaatc actcagggtc aatgccagcg cttcgttaat acagatgtag
     3901 gtgttccaca gggtagccag cagcatcctg cgatgcagat ccggaacata atggtgcagg
     3961 gcgctgactt ccgcgtttcc agactttacg aaacacggaa accgaagacc attcatgttg
     4021 ttgctcaggt cgcagacgtt ttgcagcagc agtcgcttca cgttcgctcg cgtatcggtg
     4081 attcattctg ctaaccagta aggcaacccc gccagcctag ccgggtcctc aacgacagga
     4141 gcacgatcat gcgcacccgt ggccaggacc caacgctgcc cgagatgcgc cgcgtgcggc
     4201 tgctggagat ggcggacgcg atggatatgt tctgccaagg gttggtttgc gcattcacag
     4261 ttctccgcaa gaattgattg gctccaattc ttggagtggt gaatccgtta gcgaggtgcc
     4321 gccggcttcc attcaggtcg aggtggcccg gctccatgca ccgcgacgca acgcggggag
     4381 gcagacaagg tatagggcgg cgcctacaat ccatgccaac ccgttccatg tgctcgccga
     4441 ggcggcataa atcgccgtga cgatcagcgg tccaatgatc gaagttaggc tggtaagagc
     4501 cgcgagcgat ccttgaagct gtccctgatg gtcgtcatct acctgcctgg acagcatggc
     4561 ctgcaacgcg ggcatcccga tgccgccgga agcgagaaga atcataatgg ggaaggccat
     4621 ccagcctcgc gtcgcgaacg ccagcaagac gtagcccagc gcgtcggccg ccatgccggc
     4681 gataatggcc tgcttctcgc cgaaacgttt ggtggcggga ccagtgacga aggcttgagc
     4741 gagggcgtgc aagattccga ataccgcaag cgacaggccg atcatcgtcg cgctccagcg
     4801 aaagcggtcc tcgccgaaaa tgacccagag cgctgccggc acctgtccta cgagttgcat
     4861 gataaagaag acagtcataa gtgcggcgac gatagtcatg ccccgcgccc accggaagga
     4921 gctgactggg ttgaaggctc tcaagggcat cggtcgagat cccggtgcct aatgagtgag
     4981 ctaacttaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     5041 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgcca
     5101 gggtggtttt tcttttcacc agtgagacgg gcaacagctg attgcccttc accgcctggc
     5161 cctgagagag ttgcagcaag cggtccacgc tggtttgccc cagcaggcga aaatcctgtt
     5221 tgatggtggt taacggcggg atataacatg agctgtcttc ggtatcgtcg tatcccacta
     5281 ccgagatatc cgcaccaacg cgcagcccgg actcggtaat ggcgcgcatt gcgcccagcg
     5341 ccatctgatc gttggcaacc agcatcgcag tgggaacgat gccctcattc agcatttgca
     5401 tggtttgttg aaaaccggac atggcactcc agtcgccttc ccgttccgct atcggctgaa
     5461 tttgattgcg agtgagatat ttatgccagc cagccagacg cagacgcgcc gagacagaac
     5521 ttaatgggcc cgctaacagc gcgatttgct ggtgacccaa tgcgaccaga tgctccacgc
     5581 ccagtcgcgt accgtcttca tgggagaaaa taatactgtt gatgggtgtc tggtcagaga
     5641 catcaagaaa taacgccgga acattagtgc aggcagcttc cacagcaatg gcatcctggt
     5701 catccagcgg atagttaatg atcagcccac tgacgcgttg cgcgagaaga ttgtgcaccg
     5761 ccgctttaca ggcttcgacg ccgcttcgtt ctaccatcga caccaccacg ctggcaccca
     5821 gttgatcggc gcgagattta atcgccgcga caatttgcga cggcgcgtgc agggccagac
     5881 tggaggtggc aacgccaatc agcaacgact gtttgcccgc cagttgttgt gccacgcggt
     5941 tgggaatgta attcagctcc gccatcgccg cttccacttt ttcccgcgtt ttcgcagaaa
     6001 cgtggctggc ctggttcacc acgcgggaaa cggtctgata agagacaccg gcatactctg
     6061 cgacatcgta taacgttact ggtttcacat tcaccaccct gaattgactc tcttccgggc
     6121 gctatcatgc cataccgcga aaggttttgc gccattcgat ggtgtccggg atctcgacgc
     6181 tctcccttat gcgactcctg cattaggaag cagcccagta gtaggttgag gccgttgagc
     6241 accgccgccg caaggaatgg tgcatgcaag gagatggcgc ccaacagtcc cccggccacg
     6301 gggcctgcca ccatacccac gccgaaacaa gcgctcatga gcccgaagtg gcgagcccga
     6361 tcttccccat cggtgatgtc ggcgatatag gcgccagcaa ccgcacctgt ggcgccggtg
     6421 atgccggcca cgatgcgtcc ggcgtagagg atcgagatct cgatcccgcg aaat
//
