LOCUS       sequence-201550-        6386 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  b2AR-SPARK TF component.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6386)
  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 6386)
  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-b2AR-hLOV1-TEVcs-FLAG-GAL4-V5
FEATURES             Location/Qualifiers
     source          1..6386
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     enhancer        201..504
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        505..708
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     659..679
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     753..772
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        753..771
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      785..794
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2495..2515
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQM"
     CDS             2528..2551
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2552..2989
                     /codon_start=1
                     /gene="Saccharomyces cerevisiae GAL4 (truncated)"
                     /product="DNA binding domain of the GAL4 transcriptional 
                     activator"
                     /label=GAL4 DNA binding domain
                     /translation="KLLSSIEQACDICRLKKLKCSKEKPKCAKCLKNNWECRYSPKTKR
                     SPLTRAHLTEVESRLERLEQLFLLIFPREDLDMILKMDSLQDIKALLTGLFVQDNVNKD
                     AVTDRLASVETDMPLTLRQHRISATSSSEESSNKGQRQLTVS"
     CDS             2990..3331
                     /codon_start=1
                     /gene="Saccharomyces cerevisiae GAL4 (truncated)"
                     /product="activation domain of the GAL4 transcriptional 
                     activator"
                     /label=GAL4 activation domain
                     /translation="ANFNQSGNIADSSLSFTFTNSSNGPNLITTQTNSQALSQPIASSN
                     VHDNFMNNEITASKIDDGNNSKPLSPGWTDQTAYNAFGITTGMFNTTTMDDVYNYLFDD
                     EDTPPNPKKE"
     CDS             3332..3373
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(3407..3424)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3413..3620
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3649..3789
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3649..3778
                     /label=AAV2 ITR
     rep_origin      3864..4319
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3951..3970)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4161..4182
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4336..4355)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4455..4477
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4515..4533)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4601..4705
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4706..4774,4775..5566)
                     /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(4924..4943)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5737..6325
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6226..6245
                     /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 gcgttgtaca tagtaatcaa ttacggggtc attagttcat
      181 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
      241 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
      301 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
      361 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
      421 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
      481 gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga
      541 tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg
      601 ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg
      661 caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact
      721 agagaaccca ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa
      781 gcttgccacc atggggcaac ccgggaacgg cagcgccttc ttgctggcac ccaatagaag
      841 ccatgcgccg gaccacgacg tcacgcagca gagggacgag gtgtgggtgg tgggcatggg
      901 catcgtcatg tctctcatcg tcctggccat cgtgtttggc aatgtgctgg tcatcacagc
      961 cattgccaag ttcgagcgtc tgcagacggt caccaactac ttcatcactt cactggcctg
     1021 tgctgatctg gtcatgggcc tggcagtggt gccctttggg gccgcccata ttcttatgaa
     1081 aatgtggact tttggcaact tctggtgcga gttttggact tccattgatg tgctgtgcgt
     1141 cacggccagc attgagaccc tgtgcgtgat cgcagtggat cgctactttg ccattacttc
     1201 acctttcaag taccagagcc tgctgaccaa gaataaggcc cgggtgatca ttctgatggt
     1261 gtggattgtg tcaggcctta cctccttctt gcccattcag atgcactggt accgggccac
     1321 ccaccaggaa gccatcaact gctatgccaa tgagacctgc tgtgacttct tcacgaacca
     1381 agcctatgcc attgcctctt ccatcgtgtc cttctacgtt cccctggtga tcatggtctt
     1441 cgtctactcc agggtctttc aggaggccaa aaggcagctc cagaagattg acaaatctga
     1501 gggccgcttc catgtccaga accttagcca ggtggagcag gatgggcgga cggggcatgg
     1561 actccgcaga tcttccaagt tctgcttgaa ggagcacaaa gccctcaaga cgttaggcat
     1621 catcatgggc actttcaccc tctgctggct gcccttcttc atcgttaaca ttgtgcatgt
     1681 gatccaggat aacctcatcc gtaaggaagt ttacatcctc ctaaattgga taggctatgt
     1741 caattctggt ttcaatcccc ttatctactg ccggagccca gatttcagga ttgccttcca
     1801 ggagcttctg tgcctgcgca ggtcttcttt gaaggcctat gggaatggct actccagcaa
     1861 cggcaacaca ggggagcaga gtggatatca cgtggaacag gagaaagaaa ataaactgct
     1921 gtgtgaagac ctcccaggca cggaagactt tgtgggccat caaggtactg tgcctagcga
     1981 taacattgat tcacaaggga ggaattgtag tacaaatgac tcactgctgg agctggccga
     2041 gaagctggcc ggcctggaca tcaacggtgg agctagcgag tttcgggcaa ccacactgga
     2101 acggatcgag aaaagcttcg tgattactga tccgagactg cctgacaacc caatcatttt
     2161 tgtgagcgat tccttcctgc agctgacaga atattctcgg gaagagatcc tggggcgcaa
     2221 ttgccgtttt ctgcagggac ccgagacaga ccgtgccact gttcggaaaa tcagagatgc
     2281 tattgacaac cagactgaag tgaccgttca gctgatcaat tataccaaga gcggcaagaa
     2341 gttctggaac gtgttccacc tgcagccgat gcgcgattat aagggcgacg tccagtactt
     2401 cattggcgtg cagctggatg gcaccgaacg tcttcatggc gccgctgagc gtgaggcggt
     2461 ctgcctggtc aaaaagacag cctttcagat tgctgagaac ctgtacttcc agatgggcgg
     2521 aggatccgac tacaaggatg acgacgataa gaagctactg tcttctatcg aacaagcatg
     2581 cgatatttgc cgacttaaaa agctcaagtg ctccaaagaa aaaccgaagt gcgccaagtg
     2641 tctgaagaac aactgggagt gtcgctactc tcccaaaacc aaaaggtctc cgctgactag
     2701 ggcacatctg acagaagtgg aatcaaggct agaaagactg gaacagctat ttctactgat
     2761 ttttcctcga gaagaccttg acatgatttt gaaaatggat tctttacagg atataaaagc
     2821 attgttaaca ggattatttg tacaagataa tgtgaataaa gatgccgtca cagatagatt
     2881 ggcttcagtg gagactgata tgcctctaac attgagacag catagaataa gtgcgacatc
     2941 atcatcggaa gagagtagta acaaaggtca aagacagttg actgtatcgg ccaattttaa
     3001 tcaaagtggg aatattgctg atagctcatt gtccttcact ttcactaaca gtagcaacgg
     3061 tccgaacctc ataacaactc aaacaaattc tcaagcgctt tcacaaccaa ttgcctcctc
     3121 taacgttcat gataacttca tgaataatga aatcacggct agtaaaattg atgatggtaa
     3181 taattcaaaa ccactgtcac ctggttggac ggaccaaact gcgtataacg cgtttggaat
     3241 cactacaggg atgtttaata ccactacaat ggatgatgta tataactatc tattcgatga
     3301 tgaagatacc ccaccaaacc caaaaaaaga gggcaagccg atccctaacc ctctgctggg
     3361 cctggacagc acttaagaat tcctcgagct agagctcgct gatcagcctc gactgtgcct
     3421 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
     3481 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg
     3541 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac
     3601 aatagcaggc atgctgggga ggtaaccacg tgcggaccga gcggccgcag gaacccctag
     3661 tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa
     3721 aggtcgcccg acgcccgggc tttgcccggg cggcctcagt gagcgagcga gcgcgcagct
     3781 gcctgcaggg gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc
     3841 gcatacgtca aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     3901 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ttagcgcccg ctcctttcgc
     3961 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     4021 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgattt
     4081 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     4141 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaactctat
     4201 ctcgggctat tcttttgatt tataagggat tttgccgatt tcggtctatt ggttaaaaaa
     4261 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt
     4321 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc
     4381 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca
     4441 agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg
     4501 cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat
     4561 ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     4621 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     4681 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     4741 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     4801 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     4861 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     4921 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     4981 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     5041 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     5101 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     5161 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     5221 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     5281 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     5341 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     5401 atctggagcc ggtgagcgtg gaagccgcgg tatcattgca gcactggggc cagatggtaa
     5461 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     5521 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     5581 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     5641 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     5701 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     5761 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     5821 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     5881 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     5941 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     6001 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     6061 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     6121 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     6181 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     6241 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     6301 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     6361 cttttgctgg ccttttgctc acatgt
//
