LOCUS       sequence-244556-        6275 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SPARK2 NanoLuc-beta-arrestin2-TEVp (no HA) in mammalian 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6275)
  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 6275)
  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-NanoLuc-15aa linker-beta-arrestin2-no HA-GS
            l...
FEATURES             Location/Qualifiers
     source          1..6275
                     /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      790..799
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             796..1308
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     primer_bind     complement(3301..3318)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3307..3514
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3543..3683
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3543..3672
                     /label=AAV2 ITR
     rep_origin      3758..4213
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3845..3864)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4055..4076
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4230..4249)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4349..4371
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4409..4427)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4495..4599
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4600..4668,4669..5460)
                     /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(4818..4837)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5631..6219
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6120..6139
                     /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 ccaccatggt cttcacactc gaagatttcg ttggggactg gcgacagaca
      841 gccggctaca acctggacca agtccttgaa cagggaggtg tgtccagttt gtttcagaat
      901 ctcggggtgt ccgtaactcc gatccaaagg attgtcctga gcggtgaaaa tgggctgaag
      961 atcgacatcc atgtcatcat cccgtatgaa ggtctgagcg gcgaccaaat gggccagatc
     1021 gaaaaaattt ttaaggtggt gtaccctgtg gatgatcatc actttaaggt gatcctgcac
     1081 tatggcacac tggtaatcga cggggttacg ccgaacatga tcgactattt cggacggccg
     1141 tatgaaggca tcgccgtgtt cgacggcaaa aagatcactg taacagggac cctgtggaac
     1201 ggcaacaaaa ttatcgacga gcgcctgatc aaccccgacg gctccctgct gttccgagta
     1261 accatcaacg gagtgaccgg ctggcggctg tgcgaacgca ttctggcggg cgacccacta
     1321 gtccagtgtg gtggaattgc cggatccgcc accggtgaaa aacccgggac cagggtcttc
     1381 aagaagtcga gccctaactg caagctcacc gtgtacttgg gcaagcgtga ctttgtggat
     1441 cacttggaca aagtggatcc tgtcgatggt gtggtgcttg tggatcctga ctacttgaag
     1501 gaccggaaag tgtttgtgac cctcacctgt gccttccgct atggccgaga agacctggat
     1561 gtactgggcc tgtctttccg caaagatctg ttcatcgcca cctaccaggc cttccccccc
     1621 atgcccaacc cacctcggcc ccccacccgc ctacaggacc gactgctgaa gaagttgggc
     1681 cagcacgccc accccttttt tttcacaata ccccagaatt tgccttgctc cgtcacactg
     1741 cagccaggac cggaggacac agggaaggcc tgtggagtag actttgagat tcgagccttc
     1801 tgtgccaaat ctatagaaga aaaaagccac aaaaggaact ccgtgcggct tatcatcaga
     1861 aaggtacagt ttgctcctga gacacccggc ccccagccat cagctgaaac cacacgccac
     1921 ttcctcatgt ctgaccggag gtccctgcac ctagaggctt ccctggacaa agagctgtac
     1981 taccatgggg aacccctcaa tgtcaacgtc cacgtcacca acaattctgc caaaaccgtc
     2041 aagaagatca gagtgtctgt gagacagtat gccgacattt gcctcttcag caccgcgcag
     2101 tacaagtgtc ctgtggctca gcttgaacaa gatgaccagg tgtctcccag ttccacattc
     2161 tgcaaggtgt acaccataac cccgctgctc agtgacaacc gagagaagcg tggccttgcc
     2221 cttgatgggc aactcaagca cgaagacacc aacctggctt ccagcaccat tgtgaaggag
     2281 ggagccaaca aggaggtgct gggaatccta gtatcctaca gggtcaaggt gaagctggtg
     2341 gtgtctcgag gcggggatgt ctccgtggag ctacctttcg tcctaatgca ccccaagccc
     2401 cacgaccaca tcacccttcc ccgaccccag tcagcccccc gggaaataga catccctgtg
     2461 gataccaacc tcattgaatt cgataccaac tatgccacag acgacgacat cgtgtttgag
     2521 gactttgcga ggcttcggct gaaggggatg aaggatgacg actgtgatga ccagttctgc
     2581 ggtggaagtg gatcaggcag cggtggatct ggagaaagct tgtttaaggg accacgtgat
     2641 tacaacccga tatcgagcac catttgtcat ttgacgaatg aatctgatgg gcacacaaca
     2701 tcgttgtatg gtattggatt tggtcccttc atcattacaa acaagcactt gtttagaaga
     2761 aataatggaa cactgttggt ccaatcacta catggtgtat tcaaggtcaa gaacaccacg
     2821 actttgcaac aacacctcat tgatgggagg gacatgataa ttattcgcat gcctaaggat
     2881 ttcccaccat ttcctcaaaa gctgaaattt agagagccac aaagggaaga gcgcatatgt
     2941 cttgtgacaa ccaacttcca aactaagagc atgtctagca tggtgtcaga cactagttgc
     3001 acattccctt catctgatgg catattctgg aagcattgga ttcaaaccaa ggatgggcag
     3061 tgtggcagtc cattagtatc aactagagat gggttcattg ttggtataca ctcagcatcg
     3121 aatttcacca acacaaacaa ttatttcaca agcgtgccga aaaacttcat ggaattgttg
     3181 acaaatcagg aggcgcagca gtgggttagt ggttggcgat taaatgctga ctcagtattg
     3241 tggggtggcc ataaagtttt catggtgtaa gaattcctcg agctagagct cgctgatcag
     3301 cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct
     3361 tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc
     3421 attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg
     3481 aggattggga agacaatagc aggcatgctg gggaggtaac cacgtgcgga ccgagcggcc
     3541 gcaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct cgctcactga
     3601 ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct cagtgagcga
     3661 gcgagcgcgc agctgcctgc aggggcgcct gatgcggtat tttctcctta cgcatctgtg
     3721 cggtatttca caccgcatac gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta
     3781 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccttagcg
     3841 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     3901 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     3961 aaaaaacttg atttgggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     4021 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     4081 acactcaact ctatctcggg ctattctttt gatttataag ggattttgcc gatttcggtc
     4141 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
     4201 acgtttacaa ttttatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
     4261 cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca
     4321 tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg
     4381 tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat
     4441 gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga
     4501 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     4561 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     4621 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     4681 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     4741 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     4801 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     4861 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     4921 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     4981 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     5041 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     5101 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     5161 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     5221 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     5281 tttattgctg ataaatctgg agccggtgag cgtggaagcc gcggtatcat tgcagcactg
     5341 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     5401 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     5461 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     5521 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     5581 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     5641 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     5701 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     5761 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     5821 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     5881 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     5941 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     6001 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     6061 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     6121 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     6181 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     6241 ttacggttcc tggccttttg ctggcctttt gctca
//
