LOCUS       sequence-244558-        6103 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses NanoLuc-beta-2AR-AsLOV2(V416L) in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6103)
  AUTHORS   Kim CK, Cho KF, Kim MW, Ting AY
  TITLE     Luciferase-LOV BRET enables versatile and specific transcriptional 
            readout of cellular protein-protein interactions.
  JOURNAL   eLife. 2019 Apr 3;8. pii: 43826. doi: 10.7554/eLife.43826.
  PUBMED    30942168
REFERENCE   2  (bases 1 to 6103)
  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: "eLife."; 
            date: "2019-04-3"; volume: "8. pii: 43826. doi"; pages: " 
            10.7554/eLife.43826"
COMMENT     Sequence Label: pAAV-gLuc sp-NanoLuc-HA-beta-2AR-NNES-AsLOV2(V416L)
FEATURES             Location/Qualifiers
     source          1..6103
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   6..135
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     enhancer        206..509
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        510..713
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     664..684
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     758..777
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        758..776
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      789..798
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             846..1355
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="VFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQRI
                     VLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGVT
                     PNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWR
                     LCERILA"
     CDS             1356..1382
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2667..3095
                     /codon_start=1
                     /gene="NPH1 from Avena sativa"
                     /product="light, oxygen, or voltage domain from phototropin
                     1 of Avena sativa (Herrou and Crosson, 2011)"
                     /label=LOV2
                     /note="photoswitch that undergoes a light-inducible 
                     conformational change"
                     /translation="LATTLERIEKNFLITDPRLPDNPIIFASDSFLQLTEYSREEILGR
                     NCRFLQGPETDRATVRKIRDAIDNQTEVTVQLINYTKSGKKFWNLFHLQPMRDQKGDVQ
                     YFIGVQLDGTEHVRDAAEREGVMLIKKTAENIDEAAKEL"
     primer_bind     complement(3129..3146)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3135..3342
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3371..3511
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3371..3500
                     /label=AAV2 ITR
     rep_origin      3586..4041
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3673..3692)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3883..3904
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4058..4077)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4177..4199
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4237..4255)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4323..4427
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4428..4496,4497..5288)
                     /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     complement(4646..4665)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5459..6047
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5948..5967
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 catgtcctgc aggcagctgc gcgctcgctc gctcactgag gccgcccggg cgtcgggcga
       61 cctttggtcg cccggcctca gtgagcgagc gagcgcgcag agagggagtg gccaactcca
      121 tcactagggg ttcctgcggc cgcacgcgtt gtacatagta atcaattacg gggtcattag
      181 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
      241 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
      301 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
      361 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
      421 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
      481 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
      541 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
      601 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
      661 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctctctggc
      721 taactagaga acccactgct tactggctta tcgaaattaa tacgactcac tatagggaga
      781 cccagcttgc caccatggga gtcaaagttc tgtttgccct gatctgcatc gctgtggccg
      841 aggccgtctt cacactcgaa gatttcgttg gggactggcg acagacagcc ggctacaacc
      901 tggaccaagt ccttgaacag ggaggtgtgt ccagtttgtt tcagaatctc ggggtgtccg
      961 taactccgat ccaaaggatt gtcctgagcg gtgaaaatgg gctgaagatc gacatccatg
     1021 tcatcatccc gtatgaaggt ctgagcggcg accaaatggg ccagatcgaa aaaattttta
     1081 aggtggtgta ccctgtggat gatcatcact ttaaggtgat cctgcactat ggcacactgg
     1141 taatcgacgg ggttacgccg aacatgatcg actatttcgg acggccgtat gaaggcatcg
     1201 ccgtgttcga cggcaaaaag atcactgtaa cagggaccct gtggaacggc aacaaaatta
     1261 tcgacgagcg cctgatcaac cccgacggct ccctgctgtt ccgagtaacc atcaacggag
     1321 tgaccggctg gcggctgtgc gaacgcattc tggcgtatcc gtacgacgta ccagactacg
     1381 cagggcaacc cgggaacggc agcgccttct tgctggcacc caatagaagc catgcgccgg
     1441 accacgacgt cacgcagcag agggacgagg tgtgggtggt gggcatgggc atcgtcatgt
     1501 ctctcatcgt cctggccatc gtgtttggca atgtgctggt catcacagcc attgccaagt
     1561 tcgagcgtct gcagacggtc accaactact tcatcacttc actggcctgt gctgatctgg
     1621 tcatgggcct ggcagtggtg ccctttgggg ccgcccatat tcttatgaaa atgtggactt
     1681 ttggcaactt ctggtgcgag ttttggactt ccattgatgt gctgtgcgtc acggccagca
     1741 ttgagaccct gtgcgtgatc gcagtggatc gctactttgc cattacttca cctttcaagt
     1801 accagagcct gctgaccaag aataaggccc gggtgatcat tctgatggtg tggattgtgt
     1861 caggccttac ctccttcttg cccattcaga tgcactggta ccgggccacc caccaggaag
     1921 ccatcaactg ctatgccaat gagacctgct gtgacttctt cacgaaccaa gcctatgcca
     1981 ttgcctcttc catcgtgtcc ttctacgttc ccctggtgat catggtcttc gtctactcca
     2041 gggtctttca ggaggccaaa aggcagctcc agaagattga caaatctgag ggccgcttcc
     2101 atgtccagaa ccttagccag gtggagcagg atgggcggac ggggcatgga ctccgcagat
     2161 cttccaagtt ctgcttgaag gagcacaaag ccctcaagac gttaggcatc atcatgggca
     2221 ctttcaccct ctgctggctg cccttcttca tcgttaacat tgtgcatgtg atccaggata
     2281 acctcatccg taaggaagtt tacatcctcc taaattggat aggctatgtc aattctggtt
     2341 tcaatcccct tatctactgc cggagcccag atttcaggat tgccttccag gagcttctgt
     2401 gcctgcgcag gtcttctttg aaggcctatg ggaatggcta ctccagcaac ggcaacacag
     2461 gggagcagag tggatatcac gtggaacagg agaaagaaaa taaactgctg tgtgaagacc
     2521 tcccaggcac ggaagacttt gtgggccatc aaggtactgt gcctagcgat aacattgatt
     2581 cacaagggag gaattgtagt acaaatgact cactgctgga gctggccgag aagctggccg
     2641 gcctggacat caacggtgga gctagcttgg ctactacact tgaacgtatt gagaagaact
     2701 ttctcattac tgacccaaga ttgccagata atcccattat attcgcgtcc gatagtttct
     2761 tgcagttgac agaatatagc cgtgaagaaa ttttgggaag aaactgcagg tttctacaag
     2821 gtcctgaaac tgatcgcgcg acagtgagaa aaattagaga tgccatagat aaccaaacag
     2881 aggtcactgt tcagctgatt aattatacaa agagtggtaa aaagttctgg aacctctttc
     2941 acttgcagcc tatgcgagat cagaagggag atgtccagta ctttattggg gttcagttgg
     3001 atggaactga gcatgtccga gatgctgccg agagagaggg agtcatgctg attaagaaaa
     3061 ctgcagaaaa tattgatgag gcggcaaaag aactttaaga attcctcgag ctagagctcg
     3121 ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt
     3181 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat
     3241 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag
     3301 caagggggag gattgggaag acaatagcag gcatgctggg gaggtaacca cgtgcggacc
     3361 gagcggccgc aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg
     3421 ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca
     3481 gtgagcgagc gagcgcgcag ctgcctgcag gggcgcctga tgcggtattt tctccttacg
     3541 catctgtgcg gtatttcaca ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg
     3601 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     3661 ccttagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     3721 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     3781 tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag tgggccatcg ccctgataga
     3841 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     3901 ctggaacaac actcaactct atctcgggct attcttttga tttataaggg attttgccga
     3961 tttcggtcta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca
     4021 aaatattaac gtttacaatt ttatggtgca ctctcagtac aatctgctct gatgccgcat
     4081 agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc
     4141 tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt
     4201 tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat
     4261 aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg
     4321 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     4381 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     4441 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     4501 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     4561 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     4621 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     4681 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     4741 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     4801 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     4861 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     4921 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     4981 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     5041 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     5101 ctggctggtt tattgctgat aaatctggag ccggtgagcg tggaagccgc ggtatcattg
     5161 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     5221 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     5281 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     5341 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     5401 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     5461 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     5521 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     5581 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     5641 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     5701 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     5761 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     5821 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     5881 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     5941 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     6001 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     6061 cggccttttt acggttcctg gccttttgct ggccttttgc tca
//
