LOCUS       sequence-201547-        6272 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 6272)
  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 6272)
  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-AVPR2-eLOV-TEVcs-FLAG-GAL4-V5
FEATURES             Location/Qualifiers
     source          1..6272
                     /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"
     CDS             2381..2401
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQM"
     CDS             2414..2437
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2438..2875
                     /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             2876..3217
                     /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             3218..3259
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(3293..3310)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3299..3506
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3535..3675
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3535..3664
                     /label=AAV2 ITR
     rep_origin      3750..4205
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3837..3856)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4047..4068
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4222..4241)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4341..4363
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4401..4419)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4487..4591
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4592..4660,4661..5452)
                     /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(4810..4829)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5623..6211
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6112..6131
                     /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 atgctcatgg cctctaccac aagcgctgtg cccggacatc catctcttcc
      841 aagtcttccc tccaacagct cacaggagag gcctctggat acccgggacc cacttcttgc
      901 aagagcggag ctcgccttgc tctccatcgt tttcgttgcc gtcgcgttga gcaatggtct
      961 ggtgctggcc gccctggcgc gacgaggccg ccgaggccac tgggcgccta ttcatgtctt
     1021 tatcggccat ctctgtcttg cagacctggc cgtggcgctg tttcaggtgc tccctcaact
     1081 ggcttggaaa gccaccgata gattccgggg gccagacgca ctgtgtcggg ccgtgaaata
     1141 cctgcagatg gtgggaatgt atgccagttc atatatgata cttgctatga ccttggacag
     1201 acacagggct atctgtcgcc ccatgctggc gtataggcac ggttccggag cccattggaa
     1261 caggccagtt ctggttgcgt gggctttttc cctgcttctg agcttgcccc agttgtttat
     1321 tttcgcacag agaaatgtgg agggcggatc tggcgtgacc gattgctggg cttgcttcgc
     1381 tgagccatgg ggcagaagga cctacgtgac gtggattgct ctcatggtgt tcgttgctcc
     1441 caccctgggg atcgccgcat gccaggtcct gatattcaga gaaatacacg cctccttggt
     1501 tcctgggcca agcgagcgcc ctggtggaag aagacggggt cgaaggactg gctctccagg
     1561 cgaaggcgcc cacgtatcag ctgcggtagc taagaccgta cgcatgacac tggtcatcgt
     1621 cgtcgtttac gttctttgct gggccccgtt ttttttggtg cagctctggg cggcctggga
     1681 ccccgaagca cccctggaag gtgctccatt cgttctgctc atgcttctcg cttcactgaa
     1741 tagctgtacc aatccttgga tttacgcctc tttctcctcc agtgttagct ccgagctgcg
     1801 gagtcttctc tgttgcgccc ggggcaggac ccccccaagc ctgggtccac aggacgaatc
     1861 ctgcactacc gcctccagtt ccctggccaa agacacctct tctgagctgg ccgagaagct
     1921 ggccggcctg gacatcaacg gtggagctag cggcagccgg gcgaccaccc tggaacgtat
     1981 tgaaaagagc tttgtgatta ccgatccgcg tctgccggat aacccgatta tttttgtgag
     2041 cgatagcttt ctgcagctga ccgaatatag ccgtgaagaa attctgggcc gtaactgccg
     2101 ttttctgcag ggcccggaaa ccgatcgtgc gaccgtgcgt aaaattcgtg atgcgattga
     2161 taaccagacc gaagtgaccg tgcagctgat taactatacc aaaagcggca aaaaattttg
     2221 gaacctgttt catctgcagc cgatgcgtga tcagaaaggc gatgtgcagt attttattgg
     2281 cgtgcagctg gatggtaccg aacgtgtgcg tgatgcggcg gaacgtgaag ctgtgatgct
     2341 ggtcaagaaa accgcggagg aaatcgacga agccgctaag gaaaatctct acttccagat
     2401 gggcggagga tccgactaca aggatgacga cgataagaag ctactgtctt ctatcgaaca
     2461 agcatgcgat atttgccgac ttaaaaagct caagtgctcc aaagaaaaac cgaagtgcgc
     2521 caagtgtctg aagaacaact gggagtgtcg ctactctccc aaaaccaaaa ggtctccgct
     2581 gactagggca catctgacag aagtggaatc aaggctagaa agactggaac agctatttct
     2641 actgattttt cctcgagaag accttgacat gattttgaaa atggattctt tacaggatat
     2701 aaaagcattg ttaacaggat tatttgtaca agataatgtg aataaagatg ccgtcacaga
     2761 tagattggct tcagtggaga ctgatatgcc tctaacattg agacagcata gaataagtgc
     2821 gacatcatca tcggaagaga gtagtaacaa aggtcaaaga cagttgactg tatcggccaa
     2881 ttttaatcaa agtgggaata ttgctgatag ctcattgtcc ttcactttca ctaacagtag
     2941 caacggtccg aacctcataa caactcaaac aaattctcaa gcgctttcac aaccaattgc
     3001 ctcctctaac gttcatgata acttcatgaa taatgaaatc acggctagta aaattgatga
     3061 tggtaataat tcaaaaccac tgtcacctgg ttggacggac caaactgcgt ataacgcgtt
     3121 tggaatcact acagggatgt ttaataccac tacaatggat gatgtatata actatctatt
     3181 cgatgatgaa gataccccac caaacccaaa aaaagagggc aagccgatcc ctaaccctct
     3241 gctgggcctg gacagcactt aagaattcct cgagctagag ctcgctgatc agcctcgact
     3301 gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg
     3361 gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg
     3421 agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg
     3481 gaagacaata gcaggcatgc tggggaggta accacgtgcg gaccgagcgg ccgcaggaac
     3541 ccctagtgat ggagttggcc actccctctc tgcgcgctcg ctcgctcact gaggccgggc
     3601 gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc
     3661 gcagctgcct gcaggggcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt
     3721 cacaccgcat acgtcaaagc aaccatagta cgcgccctgt agcggcgcat taagcgcggc
     3781 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccttag cgcccgctcc
     3841 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     3901 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     3961 tgatttgggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     4021 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     4081 ctctatctcg ggctattctt ttgatttata agggattttg ccgatttcgg tctattggtt
     4141 aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgtttac
     4201 aattttatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg
     4261 acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta
     4321 cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc
     4381 gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat
     4441 aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     4501 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     4561 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
     4621 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     4681 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     4741 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     4801 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
     4861 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     4921 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     4981 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     5041 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     5101 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
     5161 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     5221 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     5281 tgataaatct ggagccggtg agcgtggaag ccgcggtatc attgcagcac tggggccaga
     5341 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     5401 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
     5461 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     5521 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     5581 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     5641 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     5701 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     5761 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     5821 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     5881 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     5941 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     6001 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     6061 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     6121 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     6181 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     6241 cctggccttt tgctggcctt ttgctcacat gt
//
