LOCUS       sequence-201552-        6258 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 6258)
  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 6258)
  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-Luciferase
FEATURES             Location/Qualifiers
     source          1..6258
                     /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"
     misc_feature    2385..2973
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2438..2458)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2856..2867)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3005..3481
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3204..3223)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3521..3661
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3521..3650
                     /label=AAV2 ITR
     rep_origin      3736..4191
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3823..3842)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4033..4054
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4208..4227)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4327..4349
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4387..4405)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4473..4577
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4578..4646,4647..5438)
                     /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(4796..4815)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5609..6197
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6098..6117
                     /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 ccaccatgga agatgccaaa aacattaaga agggcccagc gccattctac ccactcgaag
      781 acgggaccgc cggcgagcag ctgcacaaag ccatgaagcg ctacgccctg gtgcccggca
      841 ccatcgcctt taccgacgca catatcgagg tggacattac ctacgccgag tacttcgaga
      901 tgagcgttcg gctggcagaa gctatgaagc gctatgggct gaatacaaac catcggatcg
      961 tggtgtgcag cgagaatagc ttgcagttct tcatgcccgt gttgggtgcc ctgttcatcg
     1021 gtgtggctgt ggccccagct aacgacatct acaacgagcg cgagctgctg aacagcatgg
     1081 gcatcagcca gcccaccgtc gtattcgtga gcaagaaagg gctgcaaaag atcctcaacg
     1141 tgcaaaagaa gctaccgatc atacaaaaga tcatcatcat ggatagcaag accgactacc
     1201 agggcttcca aagcatgtac accttcgtga cttcccattt gccacccggc ttcaacgagt
     1261 acgacttcgt gcccgagagc ttcgaccggg acaaaaccat cgccctgatc atgaacagta
     1321 gtggcagtac cggattgccc aagggcgtag ccctaccgca ccgcaccgct tgtgtccgat
     1381 tcagtcatgc ccgcgacccc atcttcggca accagatcat ccccgacacc gctatcctca
     1441 gcgtggtgcc atttcaccac ggcttcggca tgttcaccac gctgggctac ttgatctgcg
     1501 gctttcgggt cgtgctcatg taccgcttcg aggaggagct attcttgcgc agcttgcaag
     1561 actataagat tcaatctgcc ctgctggtgc ccacactatt tagcttcttc gctaagagca
     1621 ctctcatcga caagtacgac ctaagcaact tgcacgagat cgccagcggc ggggcgccgc
     1681 tcagcaagga ggtaggtgag gccgtggcca aacgcttcca cctaccaggc atccgccagg
     1741 gctacggcct gacagaaaca accagcgcca ttctgatcac ccccgaaggg gacgacaagc
     1801 ctggcgcagt aggcaaggtg gtgcccttct tcgaggctaa ggtggtggac ttggacaccg
     1861 gtaagacact gggtgtgaac cagcgcggcg agctgtgcgt ccgtggcccc atgatcatga
     1921 gcggctacgt taacaacccc gaggctacaa acgctctcat cgacaaggac ggctggctgc
     1981 acagcggcga catcgcctac tgggacgagg acgagcactt cttcatcgtg gaccggctga
     2041 agagcctgat caaatacaag ggctaccagg tagccccagc cgaactggag agcatcctgc
     2101 tgcaacaccc caacatcttc gacgccgggg tcgccggcct gcccgacgac gatgccggcg
     2161 agctgcccgc cgcagtcgtc gtgctggaac acggtaaaac catgaccgag aaggagatcg
     2221 tggactatgt ggccagccag gttacaaccg ccaagaagct gcgcggtggt gttgtgttcg
     2281 tggacgaggt gcctaaagga ctgaccggca agttggacgc ccgcaagatc cgcgagattc
     2341 tcattaaggc caagaaggac ctttgagata tcaagcttat cgataatcaa cctctggatt
     2401 acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg
     2461 gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct
     2521 cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc
     2581 aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca
     2641 ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac
     2701 tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt
     2761 ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct gctcgcctat gttgccacct
     2821 ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc
     2881 cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga
     2941 cgagtcggat ctccctttgg gccgcctccc cgcatcgata ccgagcgctg ctcgagagat
     3001 ctacgggtgg catccctgtg acccctcccc agtgcctctc ctggccctgg aagttgccac
     3061 tccagtgccc accagccttg tcctaataaa attaagttgc atcattttgt ctgactaggt
     3121 gtccttctat aatattatgg ggtggagggg ggtggtatgg agcaaggggc aagttgggaa
     3181 gacaacctgt agggcctgcg gggtctattg ggaaccaagc tggagtgcag tggcacaatc
     3241 ttggctcact gcaatctccg cctcctgggt tcaagcgatt ctcctgcctc agcctcccga
     3301 gttgttggga ttccaggcat gcatgaccag gctcagctaa tttttgtttt tttggtagag
     3361 acggggtttc accatattgg ccaggctggt ctccaactcc taatctcagg tgatctaccc
     3421 accttggcct cccaaattgc tgggattaca ggcgtgaacc actgctccct tccctgtcct
     3481 tctgattttg taggtaacca cgtgcggacc gagcggccgc aggaacccct agtgatggag
     3541 ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc
     3601 cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag
     3661 gggcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatacgt
     3721 caaagcaacc atagtacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     3781 cgcgcagcgt gaccgctaca cttgccagcg ccttagcgcc cgctcctttc gctttcttcc
     3841 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     3901 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg
     3961 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     4021 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaactct atctcgggct
     4081 attcttttga tttataaggg attttgccga tttcggtcta ttggttaaaa aatgagctga
     4141 tttaacaaaa atttaacgcg aattttaaca aaatattaac gtttacaatt ttatggtgca
     4201 ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac
     4261 ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga
     4321 ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac
     4381 gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt
     4441 agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     4501 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     4561 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     4621 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     4681 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     4741 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     4801 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     4861 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     4921 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     4981 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     5041 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     5101 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     5161 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     5221 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     5281 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     5341 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     5401 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     5461 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     5521 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     5581 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     5641 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     5701 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     5761 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     5821 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     5881 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     5941 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     6001 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     6061 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     6121 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     6181 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     6241 ggccttttgc tcacatgt
//
