LOCUS       Exported                9314 bp ds-DNA     circular SYN 24-JUN-2021
DEFINITION  Mammalian expression of 3xFlag-tagged human androgen receptor 
            Arginine mutant: AR-Arg13Lys (AR-R13K).
ACCESSION   .
VERSION     .
KEYWORDS    Flag-M4-AR-R13K
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9314)
  TITLE     Balk lab plasmids
REFERENCE   2  (bases 1 to 9314)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jun 24, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..9314
                     /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    4073..4695
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4272..4291)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(4719..4739)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4737..5034
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     4966..4985
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5088..5882
                     /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(5142..5161)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5752..5771
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6534..6668
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6571..6590)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6625..6644
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6673..6692)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6674..6692)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6706..6722)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6706..6722)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6719..6741)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6730..6746
                     /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(6754..6784)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6799..6820
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6937..6954)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7108..7696)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7188..7207)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7867..8727)
                     /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     8490..8509
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8728..8832)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      8859..9314
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8946..8965)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9156..9177
                     /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 tcaattggtg 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 ctaccctaag ccgccgtcca agacctaccg aggagctttc
     1081 cagaatctgt tccagagcgt gcgcgaagtg atccagaacc cgggccccag gcacccagag
     1141 gccgcgagcg cagcacctcc cggcgccagt ttgctgctgc agcagcagca gcagcagcag
     1201 cagcagcagc agcagcagca gcagcagcag cagcaacagc aagagactag ccccaggcag
     1261 cagcagcagc agcagggtga ggatggttct ccccaagccc atcgtagagg ccccacaggc
     1321 tacctggtcc tggatgagga acagcaacct tcacagccgc agtcggccct ggagtgccac
     1381 cccgagagag gttgcgtccc agagcctgga gccgccgtgg ccgccagcaa ggggctgccg
     1441 cagcagctgc cagcacctcc ggacgaggat gactcagctg ccccatccac gttgtccctg
     1501 ctgggcccca ctttccccgg cttaagcagc tgctccgctg accttaaaga catcctgagc
     1561 gaggccagca ccatgcaact ccttcagcaa cagcagcagg aagcagtatc cgaaggcagc
     1621 agcagcggga gagcgaggga ggcctcgggg gctcccactt cctccaagga caattactta
     1681 gggggcactt cgaccatttc tgacaacgcc aaggagttgt gtaaggcagt gtcggtgtcc
     1741 atgggcctgg gtgtggaggc gttggagcat ctgagtccag gggaacagct tcggggggat
     1801 tgcatgtacg ccccactttt gggagttcca cccgctgtgc gtcccactcc ttgtgcccca
     1861 ttggccgaat gcaaaggttc tctgctagac gacagcgcag gcaagagcac tgaagatact
     1921 gctgagtatt cccctttcaa gggaggttac accaaagggc tagaaggcga gagcctaggc
     1981 tgctctggca gcgctgcagc agggagctcc gggacacttg aactgccgtc taccctgtct
     2041 ctctacaagt ccggagcact ggacgaggca gctgcgtacc agagtcgcga ctactacaac
     2101 tttccactgg ctctggccgg accgccgccc cctccgccgc ctccccatcc ccacgctcgc
     2161 atcaagctgg agaacccgct ggactacggc agcgcctggg cggctgcggc ggcgcagtgc
     2221 cgctatgggg acctggcgag cctgcatggc gcgggtgcag cgggacccgg ttctgggtca
     2281 ccctcagccg ccgcttcctc atcctggcac actctcttca cagccgaaga aggccagttg
     2341 tatggaccgt gtggtggtgg tgggggtggt ggcggcggcg gcggcggcgg cggcggcggc
     2401 ggcggcggcg gcggcggcga ggcgggagct gtagccccct acggctacac tcggccccct
     2461 caggggctgg cgggccagga aagcgacttc accgcacctg atgtgtggta ccctggcggc
     2521 atggtgagca gagtgcccta tcccagtccc acttgtgtca aaagcgaaat gggcccctgg
     2581 atggatagct actccggacc ttacggggac atgcgtttgg agactgccag ggaccatgtt
     2641 ttgcccattg actattactt tccaccccag aagacctgcc tgatctgtgg agatgaagct
     2701 tctgggtgtc actatggagc tctcacatgt ggaagctgca aggtcttctt caaaagagcc
     2761 gctgaaggga aacagaagta cctgtgcgcc agcagaaatg attgcactat tgataaattc
     2821 cgaaggaaaa attgtccatc ttgtcgtctt cggaaatgtt atgaagcagg gatgactctg
     2881 ggagcccgga agctgaagaa acttggtaat ctgaaactac aggaggaagg agaggcttcc
     2941 agcaccacca gccccactga ggagacaacc cagaagctga cagtgtcaca cattgaaggc
     3001 tatgaatgtc agcccatctt tctgaatgtc ctggaagcca ttgagccagg tgtagtgtgt
     3061 gctggacacg acaacaacca gcccgactcc tttgcagcct tgctctctag cctcaatgaa
     3121 ctgggagaga gacagcttgt acacgtggtc aagtgggcca aggccttgcc tggcttccgc
     3181 aacttacacg tggacgacca gatggctgtc attcagtact cctggatggg gctcatggtg
     3241 tttgccatgg gctggcgatc cttcaccaat gtcaactcca ggatgctcta cttcgcccct
     3301 gatctggttt tcaatgagta ccgcatgcac aagtcccgga tgtacagcca gtgtgtccga
     3361 atgaggcacc tctctcaaga gtttggatgg ctccaaatca ccccccagga attcctgtgc
     3421 atgaaagcac tgctactctt cagcattatt ccagtggatg ggctgaaaaa tcaaaaattc
     3481 tttgatgaac ttcgaatgaa ctacatcaag gaactcgatc gtatcattgc atgcaaaaga
     3541 aaaaatccca catcctgctc aagacgcttc taccagctca ccaagctcct ggactccgtg
     3601 cagcctattg cgagagagct gcatcagttc acttttgacc tgctaatcaa gtcacacatg
     3661 gtgagcgtgg actttccgga aatgatggca gagatcatct ctgtgcaagt gcccaagatc
     3721 ctttctggga aagtcaagcc catctatttc cacacccagt gaagcattgg aaaccctatt
     3781 tccccacccc agctcatgcc ccctttcaga tgtcttctgc ctgttataac tctgcactac
     3841 tcctctgcag tgccttgggg aatttcctct attgatgtac agtctgtcat gaacatgttc
     3901 ctcaattcta tttgctgggc tttttttttc tctttctctc ctttcttttt cttcttccct
     3961 ccctatctaa ccctcccatg gcaccttcag actttgcttc ccattgtggc tcctatctgt
     4021 gttttgaatg gtgttgtatg cctttatcca gcacagtggc ggccgcgatt ccgggtggca
     4081 tccctgtgac ccctccccag tgcctctcct ggccctggaa gttgccactc cagtgcccac
     4141 cagccttgtc ctaataaaat taagttgcat cattttgtct gactaggtgt ccttctataa
     4201 tattatgggg tggagggggg tggtatggag caaggggcaa gttgggaaga caacctgtag
     4261 ggcctgcggg gtctattggg aaccaagctg gagtgcagtg gcacaatctt ggctcactgc
     4321 aatctccgcc tcctgggttc aagcgattct cctgcctcag cctcccgagt tgttgggatt
     4381 ccaggcatgc atgaccaggc tcagctaatt tttgtttttt tggtagagac ggggtttcac
     4441 catattggcc aggctggtct ccaactccta atctcaggtg atctacccac cttggcctcc
     4501 caaattgctg ggattacagg cgtgaaccac tgctcccttc cctgtccttc tgattttaaa
     4561 ataactatac cagcaggagg acgtccagac acagcatagg ctacctggcc atgcccaacc
     4621 ggtgggacat ttgagttgct tgcttggcac tgtcctctca tgcgttgggt ccactcagta
     4681 gatgcctgtt gaattgggta cgcggccagc ttggctgtgg aatgtgtgtc agttagggtg
     4741 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     4801 agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     4861 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
     4921 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     4981 cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggaat
     5041 tgatcagctt gggatctgat caagagacag gatgaggatc gtttcgcatg attgaacaag
     5101 atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg
     5161 cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc
     5221 cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag
     5281 cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca
     5341 ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat
     5401 ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata
     5461 cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac
     5521 gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc
     5581 tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg
     5641 tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg
     5701 gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta
     5761 cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg
     5821 gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct
     5881 gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga
     5941 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc
     6001 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc acctcgggct
     6061 cgatcccctc gcgagttggt tcagctgctg cctgaggctg gacgacctcg cggagttcta
     6121 ccggcagtgc aaatccgtcg gcatccagga aaccagcagc ggctatccgc gcatccatgc
     6181 ccccgaactg caggagtggg gaggcacgat ggccgctttg gtcgacccgg acgggacgct
     6241 cctgcgcctg atacagaacg aattgcttgc aggcatctca tgagtgtgtc ttcccgtttt
     6301 ccgcctgagg tcactgcgtg gatggagcgc tggcgcctgc tgcgcgacgg cgagctgctc
     6361 accacccact cgccaagctg gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     6421 ctccgccgat cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc
     6481 tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt
     6541 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     6601 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     6661 tctggatcaa ttccctatag tgagtcgtat taaattcgta atcatgtcat agctgtttcc
     6721 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     6781 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     6841 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     6901 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     6961 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     7021 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     7081 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     7141 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     7201 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     7261 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     7321 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     7381 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     7441 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     7501 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     7561 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     7621 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     7681 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     7741 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     7801 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     7861 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     7921 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     7981 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     8041 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     8101 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     8161 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     8221 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     8281 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     8341 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     8401 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     8461 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     8521 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     8581 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     8641 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     8701 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     8761 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     8821 aggggttccg cgcacatttc cccgaaaagt gccacctgac gcgccctgta gcggcgcatt
     8881 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     8941 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     9001 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     9061 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     9121 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     9181 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc
     9241 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt
     9301 aacgcttaca attt
//
