LOCUS       sequence-201549-        6398 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 6398)
  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 6398)
  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-eLOV-TEVcs-FLAG-GAL4-V5
FEATURES             Location/Qualifiers
     source          1..6398
                     /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             2507..2527
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQM"
     CDS             2540..2563
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2564..3001
                     /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             3002..3343
                     /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             3344..3385
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(3419..3436)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3425..3632
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3661..3801
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3661..3790
                     /label=AAV2 ITR
     rep_origin      3876..4331
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3963..3982)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4173..4194
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4348..4367)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4467..4489
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4527..4545)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4613..4717
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4718..4786,4787..5578)
                     /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(4936..4955)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5749..6337
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6238..6257
                     /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 agctagcggc agccgggcga ccaccctgga
     2101 acgtattgaa aagagctttg tgattaccga tccgcgtctg ccggataacc cgattatttt
     2161 tgtgagcgat agctttctgc agctgaccga atatagccgt gaagaaattc tgggccgtaa
     2221 ctgccgtttt ctgcagggcc cggaaaccga tcgtgcgacc gtgcgtaaaa ttcgtgatgc
     2281 gattgataac cagaccgaag tgaccgtgca gctgattaac tataccaaaa gcggcaaaaa
     2341 attttggaac ctgtttcatc tgcagccgat gcgtgatcag aaaggcgatg tgcagtattt
     2401 tattggcgtg cagctggatg gtaccgaacg tgtgcgtgat gcggcggaac gtgaagctgt
     2461 gatgctggtc aagaaaaccg cggaggaaat cgacgaagcc gctaaggaaa atctctactt
     2521 ccagatgggc ggaggatccg actacaagga tgacgacgat aagaagctac tgtcttctat
     2581 cgaacaagca tgcgatattt gccgacttaa aaagctcaag tgctccaaag aaaaaccgaa
     2641 gtgcgccaag tgtctgaaga acaactggga gtgtcgctac tctcccaaaa ccaaaaggtc
     2701 tccgctgact agggcacatc tgacagaagt ggaatcaagg ctagaaagac tggaacagct
     2761 atttctactg atttttcctc gagaagacct tgacatgatt ttgaaaatgg attctttaca
     2821 ggatataaaa gcattgttaa caggattatt tgtacaagat aatgtgaata aagatgccgt
     2881 cacagataga ttggcttcag tggagactga tatgcctcta acattgagac agcatagaat
     2941 aagtgcgaca tcatcatcgg aagagagtag taacaaaggt caaagacagt tgactgtatc
     3001 ggccaatttt aatcaaagtg ggaatattgc tgatagctca ttgtccttca ctttcactaa
     3061 cagtagcaac ggtccgaacc tcataacaac tcaaacaaat tctcaagcgc tttcacaacc
     3121 aattgcctcc tctaacgttc atgataactt catgaataat gaaatcacgg ctagtaaaat
     3181 tgatgatggt aataattcaa aaccactgtc acctggttgg acggaccaaa ctgcgtataa
     3241 cgcgtttgga atcactacag ggatgtttaa taccactaca atggatgatg tatataacta
     3301 tctattcgat gatgaagata ccccaccaaa cccaaaaaaa gagggcaagc cgatccctaa
     3361 ccctctgctg ggcctggaca gcacttaaga attcctcgag ctagagctcg ctgatcagcc
     3421 tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg
     3481 accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat
     3541 tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag
     3601 gattgggaag acaatagcag gcatgctggg gaggtaacca cgtgcggacc gagcggccgc
     3661 aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg
     3721 ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc
     3781 gagcgcgcag ctgcctgcag gggcgcctga tgcggtattt tctccttacg catctgtgcg
     3841 gtatttcaca ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg gcgcattaag
     3901 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccttagcgcc
     3961 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     4021 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     4081 aaaacttgat ttgggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     4141 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     4201 actcaactct atctcgggct attcttttga tttataaggg attttgccga tttcggtcta
     4261 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
     4321 gtttacaatt ttatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     4381 gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc
     4441 cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc
     4501 atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt
     4561 catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac
     4621 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     4681 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     4741 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     4801 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     4861 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     4921 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     4981 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     5041 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     5101 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     5161 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     5221 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     5281 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     5341 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     5401 tattgctgat aaatctggag ccggtgagcg tggaagccgc ggtatcattg cagcactggg
     5461 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     5521 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     5581 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     5641 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     5701 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     5761 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     5821 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     5881 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     5941 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     6001 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     6061 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     6121 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     6181 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     6241 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     6301 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     6361 acggttcctg gccttttgct ggccttttgc tcacatgt
//
