LOCUS       sequence-244567-        6329 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SPARK2 NanoLuc-beta-arrestin2-TEVp in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6329)
  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 6329)
  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-HA-GS
            link...
FEATURES             Location/Qualifiers
     source          1..6329
                     /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"
     CDS             2581..2607
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2608..2634
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(3355..3372)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3361..3568
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3597..3737
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3597..3726
                     /label=AAV2 ITR
     rep_origin      3812..4267
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3899..3918)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4109..4130
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4284..4303)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4403..4425
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4463..4481)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4549..4653
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4654..4722,4723..5514)
                     /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(4872..4891)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5685..6273
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6174..6193
                     /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 tacccatacg atgtgccaga ttacgcctat ccctatgacg tccctgatta cgctggtgga
     2641 agtggatcag gcagcggtgg atctggagaa agcttgttta agggaccacg tgattacaac
     2701 ccgatatcga gcaccatttg tcatttgacg aatgaatctg atgggcacac aacatcgttg
     2761 tatggtattg gatttggtcc cttcatcatt acaaacaagc acttgtttag aagaaataat
     2821 ggaacactgt tggtccaatc actacatggt gtattcaagg tcaagaacac cacgactttg
     2881 caacaacacc tcattgatgg gagggacatg ataattattc gcatgcctaa ggatttccca
     2941 ccatttcctc aaaagctgaa atttagagag ccacaaaggg aagagcgcat atgtcttgtg
     3001 acaaccaact tccaaactaa gagcatgtct agcatggtgt cagacactag ttgcacattc
     3061 ccttcatctg atggcatatt ctggaagcat tggattcaaa ccaaggatgg gcagtgtggc
     3121 agtccattag tatcaactag agatgggttc attgttggta tacactcagc atcgaatttc
     3181 accaacacaa acaattattt cacaagcgtg ccgaaaaact tcatggaatt gttgacaaat
     3241 caggaggcgc agcagtgggt tagtggttgg cgattaaatg ctgactcagt attgtggggt
     3301 ggccataaag ttttcatggt gtaagaattc ctcgagctag agctcgctga tcagcctcga
     3361 ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc
     3421 tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc
     3481 tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt
     3541 gggaagacaa tagcaggcat gctggggagg taaccacgtg cggaccgagc ggccgcagga
     3601 acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca ctgaggccgg
     3661 gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc
     3721 gcgcagctgc ctgcaggggc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat
     3781 ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct gtagcggcgc attaagcgcg
     3841 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgcctt agcgcccgct
     3901 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     3961 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     4021 cttgatttgg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     4081 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     4141 aactctatct cgggctattc ttttgattta taagggattt tgccgatttc ggtctattgg
     4201 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgttt
     4261 acaattttat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc
     4321 cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct
     4381 tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca
     4441 ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg
     4501 ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     4561 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     4621 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     4681 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     4741 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     4801 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     4861 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
     4921 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     4981 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     5041 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     5101 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     5161 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
     5221 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     5281 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     5341 gctgataaat ctggagccgg tgagcgtgga agccgcggta tcattgcagc actggggcca
     5401 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     5461 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     5521 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     5581 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     5641 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     5701 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     5761 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     5821 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     5881 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     5941 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     6001 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     6061 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     6121 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     6181 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     6241 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     6301 ttcctggcct tttgctggcc ttttgctca
//
