LOCUS       sequence-256762-        7348 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  NS3aH1-VPR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 7348)
  AUTHORS   Foight GW, Wang Z, Wei CT, Jr Greisen P, Warner KM, 
            Cunningham-Bryant D, Park K, Brunette TJ, Sheffler W, Baker D, Maly 
            DJ
  TITLE     Multi-input chemical control of protein dimerization for programming
            graded cellular responses.
  JOURNAL   Nat Biotechnol. 2019 Sep 9. pii: 10.1038/s41587-019-0242-8. doi: 
            10.1038/s41587-019-0242-8.
  PUBMED    31501561
REFERENCE   2  (bases 1 to 7348)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Biotechnol. 2019 Sep 9. pii: 10.1038/s41587-019-0242-8. doi: 
            10.1038/s41587-019-0242-8."
COMMENT     Sequence Label: GWF196
FEATURES             Location/Qualifiers
     source          1..7348
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     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"
     protein_bind    820..838
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    841..859
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     869..893
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             1614..1643
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1671..1820
                     /codon_start=1
                     /product="tetrameric repeat of the minimal activation 
                     domain of herpes simplex virus VP16 (Beerli et al., 1998)"
                     /label=VP64
                     /translation="DALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDF
                     DLDML"
     CDS             1827..1847
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2277..2648
                     /codon_start=1
                     /product="transcriptional activation domain of human RelA, 
                     also known as p65 (O'Shea and Perkins, 2008)"
                     /label=RelA (p65) AD
                     /translation="PTQAGEGTLSEALLQLQFDDEDLGALLGNSTDPAVFTDLASVDNS
                     EFQQLLNQGIPVAPHTTEPMLMEYPEAITRLVTGAQRPPDPAPAPLGAPGLPNGLLSGD
                     EDFSSIADMDFSALLSQISS"
     CDS             2667..3236
                     /codon_start=1
                     /product="transcriptional activation domain from the human 
                     herpesvirus 4 (Epstein-Barr virus) replication and 
                     transcription activator Rta/BRLF1 (Hardwick et al., 1992; 
                     Chavez et al., 2015)"
                     /label=Rta AD
                     /translation="RDSREGMFLPKPEAGSAISDVFEGREVCQPKRIRPFHPPGSPWAN
                     RPLPASLAPTPTGPVHEPVGSLTPAPVPQPLDPAPAVTPEASHLLEDPDEETSQAVKAL
                     REMADTVIPQKEEAAICGQMDLSHPPPRGHLDELTTTLESMTEDLNLDSPLTPELNEIL
                     DTFLNDECLLHAMHISTGLSIFDTSLF"
     primer_bind     complement(3300..3317)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3306..3530
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3663..3682)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    3814..3861
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             3869..4891
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             3869..4891
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     polyA_signal    5021..5142
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5058..5077)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5112..5131
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5191..5207)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5191..5207)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5204..5226)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5215..5231
                     /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(join(5239..5245,5246..5263,5264..5269))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5284..5305
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5422..5439)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5593..6181)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5673..5692)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6352..7143,7144..7212))
                     /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     6975..6994
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7213..7317)
                     /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 ccctatcagt gatagagatc
      841 tccctatcag tgatagagat cgtcgacgag ctcgtttagt gaaccgtcag atcgcctgga
      901 gacgccatcc acgctgtttt gacctccata gaagacaccg ggaccgatcc agcctccgga
      961 ctctagcgtt taaacttaag cttggtaccg agctcgccac catgaaaaag aaaggttctg
     1021 ttgtcatcgt tggacgcata aatctctctg gcgacaccgc atattctcag cagacgcgag
     1081 gtctggaagg gtgtcaagag acctcacaaa ccgggaggga caagaatcaa gtagaagggg
     1141 aggtccaggt ggtatctacg gcaacccaat ccttcttggc gacatctatc aatggagtcc
     1201 tgtggacggt ttatcatggt gcgggcactc ggacaattgc ctctccaaaa ggtccagtta
     1261 cccagatgta tactaatgta gacaaagatc ttgtcggatg gcaagcacca caagggtctc
     1321 ggagcctcac tccctgtacg tgtggaagtt cagaccttta tctcgtaact agacacgcgg
     1381 atgttatccc cgttaggaga agaggagata gtaggggatc cctgctttca ccacgcccaa
     1441 tatcctatct taagggttct tcaggtggtc ccttgttgtg tccggcgggt cacgcggtag
     1501 gtatattccg cgcggccgta tctactcggg gcgtggctaa ggcggtcgat ttcataccgg
     1561 ttgagtcctt ggaaacaacg atgcggagcc ccggttcagg gacaggctct ggtgagcaaa
     1621 aactcatatc tgaggaggac ttggaattct caagcgctgc aggtactagt gatgctttag
     1681 acgattttga cttagatatg cttggttcag acgcgttaga cgacttcgac ctagacatgt
     1741 taggctcaga tgcattggac gacttcgatt tagatatgtt gggctccgat gccctagatg
     1801 actttgatct agatatgcta ggtagtccca aaaagaagag gaaagtggga tcccagtatc
     1861 tgcccgacac agatgataga caccgaatcg aagagaaacg caagcgaacg tatgaaacct
     1921 tcaaatcgat catgaagaaa tcgcccttct cgggtccgac cgatcccagg cccccaccga
     1981 gaaggattgc ggtcccgtcc cgctcgtcgg ccagcgtgcc gaagcctgcg ccgcagccct
     2041 accccttcac gtcgagcctg agcacaatca attatgacga gttcccgacg atggtgttcc
     2101 cctcgggaca aatctcacaa gcctcggcgc tcgcaccagc gcctccccaa gtccttccgc
     2161 aagcgcctgc cccagcgcct gcaccggcaa tggtgtccgc cctcgcacag gcccctgcgc
     2221 ccgtccccgt gctcgcgcct ggaccgcccc aggcggtcgc tccaccggct ccgaagccga
     2281 cgcaggccgg agagggaaca ctctccgaag cacttcttca actccagttt gatgacgagg
     2341 atcttggagc actccttgga aactcgacag accctgcggt gtttaccgac ctcgcgtcag
     2401 tagataactc cgaatttcag cagcttttga accagggtat cccggtcgcg ccacatacaa
     2461 cggagcccat gttgatggaa taccccgaag caatcacgag acttgtgacg ggagcgcagc
     2521 ggcctcccga tcccgcaccc gcacctttgg gggcacctgg cctccctaac ggacttttga
     2581 gcggcgacga ggatttctcc tccatcgccg atatggattt ctcagccttg ctgtcacaga
     2641 tttccagcgg ctctggcagc ggcagccggg attccaggga agggatgttt ttgccgaagc
     2701 ctgaggccgg ctccgctatt agtgacgtgt ttgagggccg cgaggtgtgc cagccaaaac
     2761 gaatccggcc atttcatcct ccaggaagtc catgggccaa ccgcccactc cccgccagcc
     2821 tcgcaccaac accaaccggt ccagtacatg agccagtcgg gtcactgacc ccggcaccag
     2881 tccctcagcc actggatcca gcgcccgcag tgactcccga ggccagtcac ctgttggagg
     2941 atcccgatga agagacgagc caggctgtca aagcccttcg ggagatggcc gatactgtga
     3001 ttccccagaa ggaagaggct gcaatctgtg gccaaatgga cctttcccat ccgcccccaa
     3061 ggggccatct ggatgagctg acaaccacac ttgagtccat gaccgaggat ctgaacctgg
     3121 actcacccct gaccccggaa ttgaacgaga ttctggatac cttcctgaac gacgagtgcc
     3181 tcttgcatgc catgcatatc agcacaggac tgtccatctt cgacacatct ctgttttgat
     3241 aaatccagca cagtggcggc cgctcgagtc tagagggccc gtttaaaccc gctgatcagc
     3301 ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt
     3361 gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca
     3421 ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga
     3481 ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc
     3541 ggaaagaacc agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag
     3601 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     3661 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     3721 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     3781 aaaacttgat tagggtgatg gttcacgtac ctagaagttc ctattccgaa gttcctattc
     3841 tctagaaagt ataggaactt ccttggccaa aaagcctgaa ctcaccgcga cgtctgtcga
     3901 gaagtttctg atcgaaaagt tcgacagcgt ctccgacctg atgcagctct cggagggcga
     3961 agaatctcgt gctttcagct tcgatgtagg agggcgtgga tatgtcctgc gggtaaatag
     4021 ctgcgccgat ggtttctaca aagatcgtta tgtttatcgg cactttgcat cggccgcgct
     4081 cccgattccg gaagtgcttg acattgggga attcagcgag agcctgacct attgcatctc
     4141 ccgccgtgca cagggtgtca cgttgcaaga cctgcctgaa accgaactgc ccgctgttct
     4201 gcagccggtc gcggaggcca tggatgcgat cgctgcggcc gatcttagcc agacgagcgg
     4261 gttcggccca ttcggaccgc aaggaatcgg tcaatacact acatggcgtg atttcatatg
     4321 cgcgattgct gatccccatg tgtatcactg gcaaactgtg atggacgaca ccgtcagtgc
     4381 gtccgtcgcg caggctctcg atgagctgat gctttgggcc gaggactgcc ccgaagtccg
     4441 gcacctcgtg cacgcggatt tcggctccaa caatgtcctg acggacaatg gccgcataac
     4501 agcggtcatt gactggagcg aggcgatgtt cggggattcc caatacgagg tcgccaacat
     4561 cttcttctgg aggccgtggt tggcttgtat ggagcagcag acgcgctact tcgagcggag
     4621 gcatccggag cttgcaggat cgccgcggct ccgggcgtat atgctccgca ttggtcttga
     4681 ccaactctat cagagcttgg ttgacggcaa tttcgatgat gcagcttggg cgcagggtcg
     4741 atgcgacgca atcgtccgat ccggagccgg gactgtcggg cgtacacaaa tcgcccgcag
     4801 aagcgcggcc gtctggaccg atggctgtgt agaagtactc gccgatagtg gaaaccgacg
     4861 ccccagcact cgtccgaggg caaaggaata gcacgtacta cgagatttcg attccaccgc
     4921 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
     4981 ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta
     5041 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     5101 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc
     5161 gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     5221 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     5281 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     5341 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     5401 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     5461 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     5521 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     5581 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     5641 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     5701 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     5761 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     5821 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     5881 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     5941 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     6001 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     6061 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     6121 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     6181 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     6241 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     6301 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     6361 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     6421 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     6481 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     6541 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     6601 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     6661 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     6721 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     6781 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     6841 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     6901 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     6961 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     7021 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     7081 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     7141 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     7201 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     7261 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     7321 atttccccga aaagtgccac ctgacgtc
//
