LOCUS       Exported                9323 bp ds-DNA     circular SYN 25-JUN-2021
DEFINITION  Mammalian expression of 3xFlag-tagged human androgen receptor Lysine
            mutant: AR-Lys222Ala mutant (AR-K222A).
ACCESSION   .
VERSION     .
KEYWORDS    Flag-M4-AR-K222A
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 cgcggacaat
     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 tttcaaggga ggttacacca aagggctaga 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
//
