LOCUS HA-HA-AR-DBD-LBD 6596 bp ds-DNA circular SYN 25-JUN-2021
DEFINITION Mammalian expression of 2xHA-tagged human androgen receptor
trancate: AR DNA-bindind domain and Ligand-binding domain (DBD-LBD).
ACCESSION .
VERSION .
KEYWORDS HA-HA-AR-DBD-LBD
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6596)
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 6596)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Jun 25, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6596
/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(2187..2204)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 2193..2417
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 2463..2891
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2550..2569)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 2760..2781
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(2900..2920)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 2905..3234
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 3085..3220
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 3147..3166
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 3301..4095
/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(3355..3374)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 3965..3984
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
polyA_signal 4269..4390
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(4306..4325)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 4360..4379
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(4439..4455)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4439..4455)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(4452..4474)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 4463..4479
/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(4487..4517)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4532..4553
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(4670..4687)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4841..5429)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4921..4940)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5600..6460)
/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 6223..6242
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(6461..6565)
/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 tacgcggatc caatgcgttt ggagactgcc
1021 agggaccatg ttttgcccat tgactattac tttccacccc agaagacctg cctgatctgt
1081 ggagatgaag cttctgggtg tcactatgga gctctcacat gtggaagctg caaggtcttc
1141 ttcaaaagag ccgctgaagg gaaacagaag tacctgtgcg ccagcagaaa tgattgcact
1201 attgataaat tccgaaggaa aaattgtcca tcttgtcgtc ttcggaaatg ttatgaagca
1261 gggatgactc tgggagcccg gaagctgaag aaacttggta atctgaaact acaggaggaa
1321 ggagaggctt ccagcaccac cagccccact gaggagacaa cccagaagct gacagtgtca
1381 cacattgaag gctatgaatg tcagcccatc tttctgaatg tcctggaagc cattgagcca
1441 ggtgtagtgt gtgctggaca cgacaacaac cagcccgact cctttgcagc cttgctctct
1501 agcctcaatg aactgggaga gagacagctt gtacacgtgg tcaagtgggc caaggccttg
1561 cctggcttcc gcaacttaca cgtggacgac cagatggctg tcattcagta ctcctggatg
1621 gggctcatgg tgtttgccat gggctggcga tccttcacca atgtcaactc caggatgctc
1681 tacttcgccc ctgatctggt tttcaatgag taccgcatgc acaagtcccg gatgtacagc
1741 cagtgtgtcc gaatgaggca cctctctcaa gagtttggat ggctccaaat caccccccag
1801 gaattcctgt gcatgaaagc actgctactc ttcagcatta ttccagtgga tgggctgaaa
1861 aatcaaaaat tctttgatga acttcgaatg aactacatca aggaactcga tcgtatcatt
1921 gcatgcaaaa gaaaaaatcc cacatcctgc tcaagacgct tctaccagct caccaagctc
1981 ctggactccg tgcagcctat tgcgagagag ctgcatcagt tcacttttga cctgctaatc
2041 aagtcacaca tggtgagcgt ggactttccg gaaatgatgg cagagatcat ctctgtgcaa
2101 gtgcccaaga tcctttctgg gaaagtcaag cccatctatt tccacaccca gtgattctag
2161 agggcccgtt taaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt
2221 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc
2281 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg
2341 tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga
2401 tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc
2461 ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
2521 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
2581 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
2641 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
2701 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
2761 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
2821 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
2881 taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
2941 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
3001 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
3061 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
3121 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct
3181 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc
3241 ccgggagctt gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc
3301 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
3361 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
3421 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
3481 caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
3541 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
3601 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
3661 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
3721 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
3781 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac
3841 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
3901 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
3961 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
4021 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
4081 gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc
4141 tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg
4201 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg
4261 cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
4321 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
4381 atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt
4441 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
4501 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
4561 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
4621 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
4681 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
4741 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
4801 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
4861 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
4921 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
4981 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
5041 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
5101 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
5161 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
5221 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
5281 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
5341 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
5401 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
5461 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
5521 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
5581 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
5641 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
5701 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
5761 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
5821 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
5881 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
5941 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
6001 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
6061 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
6121 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
6181 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
6241 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
6301 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
6361 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
6421 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
6481 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
6541 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtc
//
