LOCUS       Exported                8512 bp ds-DNA     circular SYN 25-JUN-2021
DEFINITION  Mammalian expression of 2xHA-tagged human androgen receptor (hAR).
ACCESSION   .
VERSION     .
KEYWORDS    HA-HA-BI-AR
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8512)
  AUTHORS   Liu X, Han W, Gulla S, Simon NI, Gao Y, Cai C, Yang H, Zhang X, Liu 
            J, Balk SP, Chen S
  TITLE     Protein phosphatase 1 suppresses androgen receptor ubiquitylation 
            and degradation.
  JOURNAL   Oncotarget. 2016 Jan 12;7(2):1754-64. doi: 10.18632/oncotarget.6434.
  PUBMED    26636645
REFERENCE   2  (bases 1 to 8512)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jun 25, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8512
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             928..954
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             970..996
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(4103..4120)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4109..4333
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4379..4807
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4466..4485)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4676..4697
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4816..4836)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4821..5150
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5001..5136
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5063..5082
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5217..6011
                     /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(5271..5290)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5881..5900
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6185..6306
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6222..6241)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6276..6295
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6355..6371)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6355..6371)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6368..6390)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6379..6395
                     /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(6403..6433)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6448..6469
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6586..6603)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6757..7345)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6837..6856)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7516..8376)
                     /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     8139..8158
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8377..8481)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttccac catggcctac ccatacgacg tcccagacta cgctcagctc
      961 cagcttatgt acccctacga cgtgcccgac tacgcgggat ccatggaagt gcagttaggg
     1021 ctgggaaggg tctaccctcg gccgccgtcc aagacctacc gaggagcttt ccagaatctg
     1081 ttccagagcg tgcgcgaagt gatccagaac ccgggcccca ggcacccaga ggccgcgagc
     1141 gcagcacctc ccggcgccag tttgctgctg ctgcagcagc agcagcagca gcagcagcag
     1201 cagcagcagc agcagcagca gcagcagcag cagcagcaag agactagccc caggcagcag
     1261 cagcagcagc agggtgagga tggttctccc caagcccatc gtagaggccc cacaggctac
     1321 ctggtcctgg atgaggaaca gcaaccttca cagccgcagt cggccctgga gtgccacccc
     1381 gagagaggtt gcgtcccaga gcctggagcc gccgtggccg ccagcaaggg gctgccgcag
     1441 cagctgccag cacctccgga cgaggatgac tcagctgccc catccacgtt gtccctgctg
     1501 ggccccactt tccccggctt aagcagctgc tccgctgacc ttaaagacat cctgagcgag
     1561 gccagcacca tgcaactcct tcagcaacag cagcaggaag cagtatccga aggcagcagc
     1621 agcgggagag cgagggaggc ctcgggggct cccacttcct ccaaggacaa ttacttaggg
     1681 ggcacttcga ccatttctga caacgccaag gagttgtgta aggcagtgtc ggtgtccatg
     1741 ggcctgggtg tggaggcgtt ggagcatctg agtccagggg aacagcttcg gggggattgc
     1801 atgtacgccc cacttttggg agttccaccc gctgtgcgtc ccactccttg tgccccattg
     1861 gccgaatgca aaggttctct gctagacgac agcgcaggca agagcactga agatactgct
     1921 gagtattccc ctttcaaggg aggttacacc aaagggctag aaggcgagag cctaggctgc
     1981 tctggcagcg ctgcagcagg gagctccggg acacttgaac tgccgtctac cctgtctctc
     2041 tacaagtccg gagcactgga cgaggcagct gcgtaccaga gtcgcgacta ctacaacttt
     2101 ccactggctc tggccggacc gccgccccct ccgccgcctc cccatcccca cgctcgcatc
     2161 aagctggaga acccgctgga ctacggcagc gcctgggcgg ctgcggcggc gcagtgccgc
     2221 tatggggacc tggcgagcct gcatggcgcg ggtgcagcgg gacccggttc tgggtcaccc
     2281 tcagccgccg cttcctcatc ctggcacact ctcttcacag ccgaagaagg ccagttgtat
     2341 ggaccgtgtg gtggtggtgg gggtggtggc 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 ggccgctcga gtctagaggg
     4081 cccgtttaaa cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt
     4141 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
     4201 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
     4261 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg
     4321 gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac
     4381 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     4441 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     4501 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     4561 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     4621 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     4681 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     4741 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     4801 gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     4861 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     4921 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     4981 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     5041 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc
     5101 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg
     5161 gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga
     5221 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     5281 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     5341 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg
     5401 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     5461 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     5521 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     5581 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     5641 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     5701 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg
     5761 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     5821 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     5881 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     5941 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     6001 agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc
     6061 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     6121 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     6181 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     6241 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     6301 atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata
     6361 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     6421 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     6481 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     6541 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     6601 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     6661 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     6721 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     6781 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     6841 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     6901 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     6961 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     7021 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     7081 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     7141 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     7201 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     7261 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     7321 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     7381 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     7441 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     7501 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     7561 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     7621 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     7681 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     7741 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     7801 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     7861 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     7921 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     7981 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     8041 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     8101 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     8161 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     8221 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     8281 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     8341 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     8401 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     8461 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tc
//
