LOCUS Exported 9323 bp ds-DNA circular SYN 25-JUN-2021 DEFINITION Mammalian expression of 3xFlag-tagged human androgen receptor Lysine mutant: AR-Lys313Ala-Lys318Ala (AR-K313A-K318A). ACCESSION . VERSION . KEYWORDS Flag-M4-AR-K313A-K318A SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9323) TITLE Balk lab plasmids REFERENCE 2 (bases 1 to 9323) AUTHORS . TITLE Direct Submission JOURNAL Exported Jun 25, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9323 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 126..148 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 140..157 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 141..157 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" enhancer 318..697 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 698..901 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 852..872 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" CDS 931..996 /codon_start=1 /product="three tandem FLAG(R) epitope tags, followed by an enterokinase cleavage site" /label=3xFLAG /translation="DYKDHDGDYKDHDIDYKDDDDK" polyA_signal 4082..4704 /label=hGH poly(A) signal /note="human growth hormone polyadenylation signal" primer_bind complement(4281..4300) /label=hGH-PA-R /note="Human growth hormone terminator, reverse primer" primer_bind complement(4728..4748) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 4746..5043 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind 4975..4994 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 5097..5891 /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(5151..5170) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 5761..5780 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 6543..6677 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(6580..6599) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 6634..6653 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(6682..6701) /label=T7 /note="T7 promoter, forward primer" promoter complement(6683..6701) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(6715..6731) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(6715..6731) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(6728..6750) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 6739..6755 /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)." promoter complement(6763..6793) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 6808..6829 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(6946..6963) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(7117..7705) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(7197..7216) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(7876..8736) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 8499..8518 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(8737..8841) /gene="bla" /label=AmpR promoter rep_origin 8868..9323 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(8955..8974) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 9165..9186 /label=F1ori-F /note="F1 origin, forward primer" ORIGIN 1 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct 61 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg 121 gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctgatct atacattgaa 181 tcaatattgg caattagcca tattagtcat tggttatata gcataaatca atattggcta 241 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc 301 aatatgaccg ccatgttgac attgattatt gactagttat taatagtaat caattacggg 361 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc 421 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat 481 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc 541 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga 601 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg 661 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac 721 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt 781 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc 841 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc 901 tcgtttagtg aaccgtcaga attaaccatg gactacaaag accatgacgg tgattataaa 961 gatcatgaca tcgattacaa ggatgacgat gacaagcttc cgcggggatc catggaagtg 1021 cagttagggc tgggaagggt ctaccctcgg ccgccgtcca agacctaccg aggagctttc 1081 cagaatctgt tccagagcgt gcgcgaagtg atccagaacc cgggccccag gcacccagag 1141 gccgcgagcg cagcacctcc cggcgccagt ttgctgctgc tgcagcagca gcagcagcag 1201 cagcagcagc agcagcagca gcagcagcag cagcagcagc agcagcaaga gactagcccc 1261 aggcagcagc agcagcagca gggtgaggat ggttctcccc aagcccatcg tagaggcccc 1321 acaggctacc tggtcctgga tgaggaacag caaccttcac agccgcagtc ggccctggag 1381 tgccaccccg agagaggttg cgtcccagag cctggagccg ccgtggccgc cagcaagggg 1441 ctgccgcagc agctgccagc acctccggac gaggatgact cagctgcccc atccacgttg 1501 tccctgctgg gccccacttt ccccggctta agcagctgct ccgctgacct taaagacatc 1561 ctgagcgagg ccagcaccat gcaactcctt cagcaacagc agcaggaagc agtatccgaa 1621 ggcagcagca gcgggagagc gagggaggcc tcgggggctc ccacttcctc caaggacaat 1681 tacttagggg gcacttcgac catttctgac aacgccaagg agttgtgtaa ggcagtgtcg 1741 gtgtccatgg gcctgggtgt ggaggcgttg gagcatctga gtccagggga acagcttcgg 1801 ggggattgca tgtacgcccc acttttggga gttccacccg ctgtgcgtcc cactccttgt 1861 gccccattgg ccgaatgcaa aggttctctg ctagacgaca gcgcaggcaa gagcactgaa 1921 gatactgctg agtattcccc tttcgcggga ggttacaccg cagggctaga aggcgagagc 1981 ctaggctgct ctggcagcgc tgcagcaggg agctccggga cacttgaact gccgtctacc 2041 ctgtctctct acaagtccgg agcactggac gaggcagctg cgtaccagag tcgcgactac 2101 tacaactttc cactggctct ggccggaccg ccgccccctc cgccgcctcc ccatccccac 2161 gctcgcatca agctggagaa cccgctggac tacggcagcg cctgggcggc tgcggcggcg 2221 cagtgccgct atggggacct ggcgagcctg catggcgcgg gtgcagcggg acccggttct 2281 gggtcaccct cagccgccgc ttcctcatcc tggcacactc tcttcacagc cgaagaaggc 2341 cagttgtatg gaccgtgtgg tggtggtggg ggtggtggcg gcggcggcgg cggcggcggc 2401 ggcggcggcg gcggcggcgg cggcggcgag gcgggagctg tagcccccta cggctacact 2461 cggccccctc aggggctggc gggccaggaa agcgacttca ccgcacctga tgtgtggtac 2521 cctggcggca tggtgagcag agtgccctat cccagtccca cttgtgtcaa aagcgaaatg 2581 ggcccctgga tggatagcta ctccggacct tacggggaca tgcgtttgga gactgccagg 2641 gaccatgttt tgcccattga ctattacttt ccaccccaga agacctgcct gatctgtgga 2701 gatgaagctt ctgggtgtca ctatggagct ctcacatgtg gaagctgcaa ggtcttcttc 2761 aaaagagccg ctgaagggaa acagaagtac ctgtgcgcca gcagaaatga ttgcactatt 2821 gataaattcc gaaggaaaaa ttgtccatct tgtcgtcttc ggaaatgtta tgaagcaggg 2881 atgactctgg gagcccggaa gctgaagaaa cttggtaatc tgaaactaca ggaggaagga 2941 gaggcttcca gcaccaccag ccccactgag gagacaaccc agaagctgac agtgtcacac 3001 attgaaggct atgaatgtca gcccatcttt ctgaatgtcc tggaagccat tgagccaggt 3061 gtagtgtgtg ctggacacga caacaaccag cccgactcct ttgcagcctt gctctctagc 3121 ctcaatgaac tgggagagag acagcttgta cacgtggtca agtgggccaa ggccttgcct 3181 ggcttccgca acttacacgt ggacgaccag atggctgtca ttcagtactc ctggatgggg 3241 ctcatggtgt ttgccatggg ctggcgatcc ttcaccaatg tcaactccag gatgctctac 3301 ttcgcccctg atctggtttt caatgagtac cgcatgcaca agtcccggat gtacagccag 3361 tgtgtccgaa tgaggcacct ctctcaagag tttggatggc tccaaatcac cccccaggaa 3421 ttcctgtgca tgaaagcact gctactcttc agcattattc cagtggatgg gctgaaaaat 3481 caaaaattct ttgatgaact tcgaatgaac tacatcaagg aactcgatcg tatcattgca 3541 tgcaaaagaa aaaatcccac atcctgctca agacgcttct accagctcac caagctcctg 3601 gactccgtgc agcctattgc gagagagctg catcagttca cttttgacct gctaatcaag 3661 tcacacatgg tgagcgtgga ctttccggaa atgatggcag agatcatctc tgtgcaagtg 3721 cccaagatcc tttctgggaa agtcaagccc atctatttcc acacccagtg aagcattgga 3781 aaccctattt ccccacccca gctcatgccc cctttcagat gtcttctgcc tgttataact 3841 ctgcactact cctctgcagt gccttgggga atttcctcta ttgatgtaca gtctgtcatg 3901 aacatgttcc tcaattctat ttgctgggct ttttttttct ctttctctcc tttctttttc 3961 ttcttccctc cctatctaac cctcccatgg caccttcaga ctttgcttcc cattgtggct 4021 cctatctgtg ttttgaatgg tgttgtatgc ctttatccag cacagtggcg gccgcgattc 4081 cgggtggcat ccctgtgacc cctccccagt gcctctcctg gccctggaag ttgccactcc 4141 agtgcccacc agccttgtcc taataaaatt aagttgcatc attttgtctg actaggtgtc 4201 cttctataat attatggggt ggaggggggt ggtatggagc aaggggcaag ttgggaagac 4261 aacctgtagg gcctgcgggg tctattggga accaagctgg agtgcagtgg cacaatcttg 4321 gctcactgca atctccgcct cctgggttca agcgattctc ctgcctcagc ctcccgagtt 4381 gttgggattc caggcatgca tgaccaggct cagctaattt ttgttttttt ggtagagacg 4441 gggtttcacc atattggcca ggctggtctc caactcctaa tctcaggtga tctacccacc 4501 ttggcctccc aaattgctgg gattacaggc gtgaaccact gctcccttcc ctgtccttct 4561 gattttaaaa taactatacc agcaggagga cgtccagaca cagcataggc tacctggcca 4621 tgcccaaccg gtgggacatt tgagttgctt gcttggcact gtcctctcat gcgttgggtc 4681 cactcagtag atgcctgttg aattgggtac gcggccagct tggctgtgga atgtgtgtca 4741 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct 4801 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca 4861 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc 4921 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta 4981 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt 5041 tggaggaatt gatcagcttg ggatctgatc aagagacagg atgaggatcg tttcgcatga 5101 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct 5161 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc 5221 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg 5281 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg 5341 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc 5401 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc 5461 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg 5521 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc 5581 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg 5641 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc 5701 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag 5761 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg 5821 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg 5881 agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc 5941 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt 6001 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca 6061 cctcgggctc gatcccctcg cgagttggtt cagctgctgc ctgaggctgg acgacctcgc 6121 ggagttctac cggcagtgca aatccgtcgg catccaggaa accagcagcg gctatccgcg 6181 catccatgcc cccgaactgc aggagtgggg aggcacgatg gccgctttgg tcgacccgga 6241 cgggacgctc ctgcgcctga tacagaacga attgcttgca ggcatctcat gagtgtgtct 6301 tcccgttttc cgcctgaggt cactgcgtgg atggagcgct ggcgcctgct gcgcgacggc 6361 gagctgctca ccacccactc gccaagctgg aaccgtaaaa aggccgcgtt gctggcgttt 6421 ttccataggc tccgccgatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt 6481 taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg 6541 ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca 6601 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat 6661 cttatcatgt ctggatcaat tccctatagt gagtcgtatt aaattcgtaa tcatgtcata 6721 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag 6781 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg 6841 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca 6901 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc 6961 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg 7021 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa 7081 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga 7141 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag 7201 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct 7261 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg 7321 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc 7381 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt 7441 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta 7501 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac 7561 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc 7621 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat 7681 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc 7741 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt 7801 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta 7861 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct 7921 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg 7981 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga 8041 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt 8101 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt 8161 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt 8221 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat 8281 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc 8341 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc 8401 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat 8461 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag 8521 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt 8581 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc 8641 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa 8701 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg 8761 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa 8821 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg cgccctgtag 8881 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag 8941 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt 9001 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca 9061 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata 9121 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca 9181 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc 9241 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa 9301 caaaatatta acgcttacaa ttt //