LOCUS       sequence-201551-        5343 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  SPARK reporter gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5343)
  AUTHORS   Kim MW, Wang W, Sanchez MI, Coukos R, von Zastrow M, Ting AY
  TITLE     Time-gated detection of protein-protein interactions with 
            transcriptional readout.
  JOURNAL   Elife. 2017 Nov 30;6. doi: 10.7554/eLife.30233.
  PUBMED    29189201
REFERENCE   2  (bases 1 to 5343)
  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: "2017-11-30"; volume: "6. doi"; pages: " 10.7554/eLife.30233"
COMMENT     Sequence Label: pAAV-UAS-CITRINE
FEATURES             Location/Qualifiers
     source          1..5343
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     5'UTR           619..661
                     /label=Xenopus globin 5'-UTR
                     /note="translational enhancer from the 5'-UTR of the major 
                     beta-globin gene of Xenopus laevis"
     regulatory      720..729
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(726..728,729..731,732..1442)
                     /codon_start=1
                     /product="enhanced variant of YFP (Heikal et al., 2001)"
                     /label=Citrine
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(771..792)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1032..1051)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1379..1400
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    1470..2058
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1523..1543)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1941..1952)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2090..2566
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2289..2308)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   2606..2746
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2606..2735
                     /label=AAV2 ITR
     rep_origin      2821..3276
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2908..2927)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3118..3139
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3293..3312)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3412..3434
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3472..3490)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3558..3662
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3663..3731,3732..4523)
                     /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(3881..3900)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4694..5282
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5183..5202
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtgcaaaa agcttaggcc tccaaggcgg agtactgtcc
      181 tccgggctgg cggagtactg tcctccggca aggtcggagt actgtcctcc gacactagag
      241 gtcggagtac tgtcctccga cgcaaggcgg agtactgtcc tccgggctgc ggagtactgt
      301 cctccggcaa ggtcggagta ctgtcctccg acactagagg tcggagtact gtcctccgac
      361 gcaaggtcgg agtactgtcc tccgacacta gaggtcggag tactgtcctc cgacgcaagg
      421 tcggagtact gtcctccgac actagaggtc ggagtactgt cctccgacgc aaggcggagt
      481 actgtcctcc gggctggcgg agtactgtcc tccggcaagg gtcgactcta gagggtatat
      541 aatggatccc atcgcgtctc agcctcactt tgagctcctc cacacgaatt ccccgatacc
      601 gtcgattcaa ggagcttgct tgttcttttt gcagaagctc agaataaacg ctcaactttg
      661 gcagatctac caagcttggt accgagctcg gatccactag tgatccccgg gtaccggtcg
      721 ccaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc
      781 tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca
      841 cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc
      901 ccaccctcgt gaccaccttc ggctacggcc tgatgtgctt cgcccgctac cccgaccaca
      961 tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca
     1021 tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca
     1081 ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg
     1141 ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga
     1201 agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc
     1261 tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca
     1321 accactacct gagctaccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca
     1381 tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca
     1441 aggacctttg agatatcaag cttatcgata atcaacctct ggattacaaa atttgtgaaa
     1501 gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa
     1561 tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat
     1621 cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt
     1681 gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc
     1741 tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc
     1801 ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg
     1861 ggaaatcatc gtcctttcct tggctgctcg cctatgttgc cacctggatt ctgcgcggga
     1921 cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc
     1981 tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc
     2041 tttgggccgc ctccccgcat cgataccgag cgctgctcga gagatctacg ggtggcatcc
     2101 ctgtgacccc tccccagtgc ctctcctggc cctggaagtt gccactccag tgcccaccag
     2161 ccttgtccta ataaaattaa gttgcatcat tttgtctgac taggtgtcct tctataatat
     2221 tatggggtgg aggggggtgg tatggagcaa ggggcaagtt gggaagacaa cctgtagggc
     2281 ctgcggggtc tattgggaac caagctggag tgcagtggca caatcttggc tcactgcaat
     2341 ctccgcctcc tgggttcaag cgattctcct gcctcagcct cccgagttgt tgggattcca
     2401 ggcatgcatg accaggctca gctaattttt gtttttttgg tagagacggg gtttcaccat
     2461 attggccagg ctggtctcca actcctaatc tcaggtgatc tacccacctt ggcctcccaa
     2521 attgctggga ttacaggcgt gaaccactgc tcccttccct gtccttctga ttttgtaggt
     2581 aaccacgtgc ggaccgagcg gccgcaggaa cccctagtga tggagttggc cactccctct
     2641 ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt
     2701 gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcgg
     2761 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag caaccatagt
     2821 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     2881 ctacacttgc cagcgcctta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     2941 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     3001 gtgctttacg gcacctcgac cccaaaaaac ttgatttggg tgatggttca cgtagtgggc
     3061 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     3121 gactcttgtt ccaaactgga acaacactca actctatctc gggctattct tttgatttat
     3181 aagggatttt gccgatttcg gtctattggt taaaaaatga gctgatttaa caaaaattta
     3241 acgcgaattt taacaaaata ttaacgttta caattttatg gtgcactctc agtacaatct
     3301 gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct
     3361 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
     3421 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga
     3481 tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca
     3541 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     3601 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     3661 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     3721 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     3781 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     3841 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     3901 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     3961 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     4021 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     4081 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     4141 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     4201 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     4261 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     4321 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     4381 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     4441 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     4501 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     4561 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     4621 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     4681 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     4741 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     4801 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     4861 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     4921 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     4981 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     5041 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     5101 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     5161 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     5221 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     5281 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     5341 tgt
//
