LOCUS       sequence-244557-        6101 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses beta-2AR-NanoLuc-AsLOV2(V416L) in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6101)
  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 6101)
  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-HA-beta-2AR-NNES-NanoLuc-15aa 
            linker-AsLOV2(V...
FEATURES             Location/Qualifiers
     source          1..6101
                     /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"
     CDS             799..825
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2110..2619
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="VFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQRI
                     VLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGVT
                     PNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWR
                     LCERILA"
     CDS             2665..3093
                     /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(3127..3144)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3133..3340
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3369..3509
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3369..3498
                     /label=AAV2 ITR
     rep_origin      3584..4039
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3671..3690)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3881..3902
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4056..4075)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4175..4197
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4235..4253)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4321..4425
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4426..4494,4495..5286)
                     /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(4644..4663)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5457..6045
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5946..5965
                     /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 cccaagcttg ccaccatgta tccgtacgac gtaccagact acgcagggca acccgggaac
      841 ggcagcgcct tcttgctggc acccaataga agccatgcgc cggaccacga cgtcacgcag
      901 cagagggacg aggtgtgggt ggtgggcatg ggcatcgtca tgtctctcat cgtcctggcc
      961 atcgtgtttg gcaatgtgct ggtcatcaca gccattgcca agttcgagcg tctgcagacg
     1021 gtcaccaact acttcatcac ttcactggcc tgtgctgatc tggtcatggg cctggcagtg
     1081 gtgccctttg gggccgccca tattcttatg aaaatgtgga cttttggcaa cttctggtgc
     1141 gagttttgga cttccattga tgtgctgtgc gtcacggcca gcattgagac cctgtgcgtg
     1201 atcgcagtgg atcgctactt tgccattact tcacctttca agtaccagag cctgctgacc
     1261 aagaataagg cccgggtgat cattctgatg gtgtggattg tgtcaggcct tacctccttc
     1321 ttgcccattc agatgcactg gtaccgggcc acccaccagg aagccatcaa ctgctatgcc
     1381 aatgagacct gctgtgactt cttcacgaac caagcctatg ccattgcctc ttccatcgtg
     1441 tccttctacg ttcccctggt gatcatggtc ttcgtctact ccagggtctt tcaggaggcc
     1501 aaaaggcagc tccagaagat tgacaaatct gagggccgct tccatgtcca gaaccttagc
     1561 caggtggagc aggatgggcg gacggggcat ggactccgca gatcttccaa gttctgcttg
     1621 aaggagcaca aagccctcaa gacgttaggc atcatcatgg gcactttcac cctctgctgg
     1681 ctgcccttct tcatcgttaa cattgtgcat gtgatccagg ataacctcat ccgtaaggaa
     1741 gtttacatcc tcctaaattg gataggctat gtcaattctg gtttcaatcc ccttatctac
     1801 tgccggagcc cagatttcag gattgccttc caggagcttc tgtgcctgcg caggtcttct
     1861 ttgaaggcct atgggaatgg ctactccagc aacggcaaca caggggagca gagtggatat
     1921 cacgtggaac aggagaaaga aaataaactg ctgtgtgaag acctcccagg cacggaagac
     1981 tttgtgggcc atcaaggtac tgtgcctagc gataacattg attcacaagg gaggaattgt
     2041 agtacaaatg actcactgct ggagctggcc gagaagctgg ccggcctgga catcaacggt
     2101 ggagctagcg tcttcacact cgaagatttc gttggggact ggcgacagac agccggctac
     2161 aacctggacc aagtccttga acagggaggt gtgtccagtt tgtttcagaa tctcggggtg
     2221 tccgtaactc cgatccaaag gattgtcctg agcggtgaaa atgggctgaa gatcgacatc
     2281 catgtcatca tcccgtatga aggtctgagc ggcgaccaaa tgggccagat cgaaaaaatt
     2341 tttaaggtgg tgtaccctgt ggatgatcat cactttaagg tgatcctgca ctatggcaca
     2401 ctggtaatcg acggggttac gccgaacatg atcgactatt tcggacggcc gtatgaaggc
     2461 atcgccgtgt tcgacggcaa aaagatcact gtaacaggga ccctgtggaa cggcaacaaa
     2521 attatcgacg agcgcctgat caaccccgac ggctccctgc tgttccgagt aaccatcaac
     2581 ggagtgaccg gctggcggct gtgcgaacgc attctggcgg gcgacccact agtccagtgt
     2641 ggtggaattg ccggatccgc caccttggct actacacttg aacgtattga gaagaacttt
     2701 ctcattactg acccaagatt gccagataat cccattatat tcgcgtccga tagtttcttg
     2761 cagttgacag aatatagccg tgaagaaatt ttgggaagaa actgcaggtt tctacaaggt
     2821 cctgaaactg atcgcgcgac agtgagaaaa attagagatg ccatagataa ccaaacagag
     2881 gtcactgttc agctgattaa ttatacaaag agtggtaaaa agttctggaa cctctttcac
     2941 ttgcagccta tgcgagatca gaagggagat gtccagtact ttattggggt tcagttggat
     3001 ggaactgagc atgtccgaga tgctgccgag agagagggag tcatgctgat taagaaaact
     3061 gcagaaaata ttgatgaggc ggcaaaagaa ctttaagaat tcctcgagct agagctcgct
     3121 gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc
     3181 cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg
     3241 catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca
     3301 agggggagga ttgggaagac aatagcaggc atgctgggga ggtaaccacg tgcggaccga
     3361 gcggccgcag gaacccctag tgatggagtt ggccactccc tctctgcgcg ctcgctcgct
     3421 cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg cggcctcagt
     3481 gagcgagcga gcgcgcagct gcctgcaggg gcgcctgatg cggtattttc tccttacgca
     3541 tctgtgcggt atttcacacc gcatacgtca aagcaaccat agtacgcgcc ctgtagcggc
     3601 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     3661 ttagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     3721 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     3781 gaccccaaaa aacttgattt gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     3841 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     3901 ggaacaacac tcaactctat ctcgggctat tcttttgatt tataagggat tttgccgatt
     3961 tcggtctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
     4021 atattaacgt ttacaatttt atggtgcact ctcagtacaa tctgctctga tgccgcatag
     4081 ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc
     4141 ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt
     4201 tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct atttttatag
     4261 gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg
     4321 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     4381 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     4441 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     4501 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     4561 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     4621 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     4681 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     4741 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     4801 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     4861 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     4921 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     4981 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     5041 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     5101 ggctggttta ttgctgataa atctggagcc ggtgagcgtg gaagccgcgg tatcattgca
     5161 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     5221 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     5281 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     5341 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     5401 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     5461 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     5521 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     5581 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     5641 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     5701 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     5761 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     5821 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     5881 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     5941 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     6001 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     6061 gcctttttac ggttcctggc cttttgctgg ccttttgctc a
//
