LOCUS       sequence-340589-        7028 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  firefly luciferase reporter vector with 12x clustered TEAD binding 
            sites.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7028)
  AUTHORS   Ma X, Mandausch FJ, Wu Y, Sahoo VK, Ma W, Leoni G, Hostiuc M, 
            Wintgens JP, Qiu J, Kannaiyan N, Rossner MJ, Wehr MC
  TITLE     Comprehensive split TEV based protein-protein interaction screening 
            reveals TAOK2 as a key modulator of Hippo signalling to limit 
            growth.
  JOURNAL   Cell Signal. 2024 Jan;113:110917. doi: 
            10.1016/j.cellsig.2023.110917. Epub 2023 Oct 7.
  PUBMED    37813295
REFERENCE   2  (bases 1 to 7028)
  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: "Cell 
            Signal."; date: "2024-01"; volume: "113:110917. doi"; pages: " 
            10.1016/j.cellsig.2023.110917. Epub 2023 Oct 7"
COMMENT     Sequence Label: pAAV_12xTEAD_luc2
FEATURES             Location/Qualifiers
     source          1..7028
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   4..133
                     /label=AAV2 ITR
     primer_bind     complement(200..219)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     protein_bind    484..508
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    join(519..525,526..527,528..534)
                     /label=2xGTIIC
                     /bound_moiety="TEF-1 family (TEAD) proteins"
                     /note="dimerized GTIIC motif from the SV40 enhancer, with 
                     high affinity for TEF-1 family (TEAD) proteins (Mahoney et 
                     al., 2005)"
     protein_bind    join(551..557,558..559,560..566)
                     /label=2xGTIIC
                     /bound_moiety="TEF-1 family (TEAD) proteins"
                     /note="dimerized GTIIC motif from the SV40 enhancer, with 
                     high affinity for TEF-1 family (TEAD) proteins (Mahoney et 
                     al., 2005)"
     protein_bind    join(583..589,590..591,592..598)
                     /label=2xGTIIC
                     /bound_moiety="TEF-1 family (TEAD) proteins"
                     /note="dimerized GTIIC motif from the SV40 enhancer, with 
                     high affinity for TEF-1 family (TEAD) proteins (Mahoney et 
                     al., 2005)"
     protein_bind    join(644..650,651..652,653..659)
                     /label=2xGTIIC
                     /bound_moiety="TEF-1 family (TEAD) proteins"
                     /note="dimerized GTIIC motif from the SV40 enhancer, with 
                     high affinity for TEF-1 family (TEAD) proteins (Mahoney et 
                     al., 2005)"
     protein_bind    join(676..682,683..684,685..691)
                     /label=2xGTIIC
                     /bound_moiety="TEF-1 family (TEAD) proteins"
                     /note="dimerized GTIIC motif from the SV40 enhancer, with 
                     high affinity for TEF-1 family (TEAD) proteins (Mahoney et 
                     al., 2005)"
     protein_bind    join(708..714,715..716,717..723)
                     /label=2xGTIIC
                     /bound_moiety="TEF-1 family (TEAD) proteins"
                     /note="dimerized GTIIC motif from the SV40 enhancer, with 
                     high affinity for TEF-1 family (TEAD) proteins (Mahoney et 
                     al., 2005)"
     protein_bind    complement(730..750)
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     protein_bind    891..911
                     /gene="mutant version of attB"
                     /label=attB3
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     promoter        938..956
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1019..1038)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1020..1038)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    complement(1062..1086)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      1165..1174
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1171..2823
                     /codon_start=1
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="synthetic luc2 version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTITEKEIVDYVASQATTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     misc_feature    2851..3439
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2904..2924)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3322..3333)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3466..3483)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3472..3696
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        complement(3906..4235)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3920..4055
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4220..4240
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(4481..5069)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4561..4580)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5240..6031,6032..6100))
                     /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     5863..5882
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6101..6205)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6273..6291
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6329..6351)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6451..6470
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6684..6703
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        6993..7011
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     6993..7010
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 tagctgcgcg ctcgctcgct cactgaggcc gcccgggcaa agcccgggcg tcgggcgacc
       61 tttggtcgcc cggcctcagt gagcgagcga gcgcgcagag agggagtggc caactccatc
      121 actaggggtt ccttgtagtt aatgattaac ccgccatgct acttatctac gtagccatgc
      181 tctaggtacc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga
      241 gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa
      301 catcctgggg cacaagctgg agtacaacta caacagccac aacgtccata tcatggccga
      361 caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag
      421 cgtgcagctc gccgaccact catatgtact cgagttgtcg acggtatcga taagcttgat
      481 atcacaagtt tgtacaaaaa agcaggctaa gcttgcagtg gaatgtgtgg aatgtgtctc
      541 aagcttgcag tggaatgtgt ggaatgtgtc tcaagcttgc agtggaatgt gtggaatgtg
      601 tctcacgcgt ggcctcgcct gcaggaggcg gccaagcttg cagtggaatg tgtggaatgt
      661 gtctcaagct tgcagtggaa tgtgtggaat gtgtctcaag cttgcagtgg aatgtgtgga
      721 atgtgtctcc aacttttcta tacaaagttg gcaagtgcag gtgccagaac atttctctgg
      781 cctaactgga agacgctagc ggggggctat aaaagggggt gggggcgttc gtcctcactc
      841 tagatctgcg atctaagtaa gcttggcaat ccggtactgt tggtaaagcc caactttgta
      901 taataaagtt gagctagttg ctaagtctgc cgagtagaat taaccctcac taaagggtag
      961 gtgacactat tctattacat caatcatcta agtgcgtctt ctattactta cctttaatcc
     1021 ctatagtgag tcgtattagt agacgcgagt caatgcatgt acgacagctt tcttgtacaa
     1081 agtggtgata tcgaattcct gcagcccggg ggatccgccc gggctagagc ggccgccacc
     1141 gcggtggagc tcgcgactag tgacgccacc atggaagatg ccaaaaacat taagaagggc
     1201 ccagcgccat tctacccact cgaagacggg accgccggcg agcagctgca caaagccatg
     1261 aagcgctacg ccctggtgcc tggcaccatc gcctttaccg acgcacatat cgaggtggac
     1321 attacctacg ccgagtactt cgagatgagc gttcggctgg cagaagctat gaagcgctat
     1381 gggctgaata caaaccatcg gatcgtggtg tgcagcgaga atagcttgca gttcttcatg
     1441 cccgtgttgg gtgccctgtt catcggtgtg gctgtggccc cagctaacga catctacaac
     1501 gagcgcgagc tgctgaacag catgggcatc agccagccca ccgtcgtatt cgtgagcaag
     1561 aaagggctgc aaaagatcct caacgtgcaa aagaagctac cgatcataca aaagatcatc
     1621 atcatggata gcaagaccga ctaccagggc ttccaaagca tgtacacctt cgtgacttcc
     1681 catttgccac ccggcttcaa cgagtacgac ttcgtgcccg agagcttcga ccgggacaaa
     1741 accatcgccc tgatcatgaa cagtagtggc agtaccggat tgcccaaggg cgtagcccta
     1801 ccgcaccgca ccgcttgtgt ccgattcagt catgcccgcg accccatctt cggcaaccag
     1861 atcatccccg acaccgctat cctcagcgtg gtgccatttc accacggctt cggcatgttc
     1921 accacgctgg gctacttgat ctgcggcttt cgggtcgtgc tcatgtaccg cttcgaggag
     1981 gagctattct tgcgcagctt gcaagactat aagattcaat ctgccctgct ggtgcccaca
     2041 ctatttagct tcttcgctaa gagcactctc atcgacaagt acgacctaag caacttgcac
     2101 gagatcgcca gcggcggggc gccgctcagc aaggaggtag gtgaggccgt ggccaaacgc
     2161 ttccacctac caggcatccg ccagggctac ggcctgacag aaacaaccag cgccattctg
     2221 atcacccccg aaggggacga caagcctggc gcagtaggca aggtggtgcc cttcttcgag
     2281 gctaaggtgg tggacttgga caccggtaag acactgggtg tgaaccagcg cggcgagctg
     2341 tgcgtccgtg gccccatgat catgagcggc tacgttaaca accccgaggc tacaaacgct
     2401 ctcatcgaca aggacggctg gctgcacagc ggcgacatcg cctactggga cgaggacgag
     2461 cacttcttca tcgtggaccg gctgaagagc ctgatcaaat acaagggcta ccaggtagcc
     2521 ccagccgaac tggagagcat cctgctgcaa caccccaaca tcttcgacgc cggggtcgcc
     2581 ggcctgcccg acgacgatgc cggcgagctg cccgccgcag tcgtcgtgct ggaacacggt
     2641 aaaaccataa ccgagaagga gatcgtggac tatgtggcca gccaggctac aaccgccaag
     2701 aagctgcgcg gtggtgttgt gttcgtggac gaggtgccta aaggactgac cggcaagttg
     2761 gacgcccgca agatccgcga gattctcatt aaggccaaga agggcggcaa gatcgccgtg
     2821 taattcgaat tcgatatcaa gcttatcgat aatcaacctc tggattacaa aatttgtgaa
     2881 agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta
     2941 atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa
     3001 tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg
     3061 tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc
     3121 ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc
     3181 cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg
     3241 gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg ccacctggat tctgcgcggg
     3301 acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg
     3361 ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc
     3421 ctttgggccg cctccccgca tcgataccgt cgactcgctg atcagcctcg actgtgcctt
     3481 ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg
     3541 ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt
     3601 gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca
     3661 atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct
     3721 ggggctcgac tagagcatgg ctacgtagat aagtagcatg gcgggttaat cattaactac
     3781 aaggaacccc tagtgatgga gttggccact ccctctctgc gcgctcgctc gctcactgag
     3841 gccgggcgac caaaggtcgc ccgacgcccg ggcggcctca gtgagcgagc gagcgcgcag
     3901 agctttttgc aaaagcctag gcctccaaaa aagcctcctc actacttctg gaatagctca
     3961 gaggccgagg cggcctcggc ctctgcataa ataaaaaaaa ttagtcagcc atggggcgga
     4021 gaatgggcgg aactgggcgg agttaggggc gggatgggcg gagttagggg cgggactatg
     4081 gttgctgact aattgagatg catgctttgc atacttctgc ctgctgggga gcctggggac
     4141 tttccacacc tggttgctga ctaattgaga tgcatgcttt gcatacttct gcctgctggg
     4201 gagcctgggg actttccaca ccctaactga cacacattcc acagctgcat taatgaatcg
     4261 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     4321 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     4381 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     4441 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     4501 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     4561 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     4621 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     4681 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     4741 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     4801 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     4861 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     4921 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     4981 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     5041 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     5101 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     5161 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     5221 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     5281 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     5341 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     5401 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     5461 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     5521 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     5581 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     5641 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     5701 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     5761 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     5821 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     5881 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     5941 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     6001 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     6061 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     6121 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     6181 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag
     6241 aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc
     6301 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
     6361 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
     6421 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
     6481 accattcgac gctctccctt atgcgactcc tgcattagga agcagcccag tagtaggttg
     6541 aggccgttga gcaccgccgc cgcaaggaat ggtgcatgca aggagatggc gcccaacagt
     6601 cccccggcca cggggcctgc caccataccc acgccgaaac aagcgctcat gagcccgaag
     6661 tggcgagccc gatcttcccc atcggtgatg tcggcgatat aggcgccagc aaccgcacct
     6721 gtggcgccgg tgatgccggc cacgatgcgt ccggcgtaga ggatctggct agcgatgacc
     6781 ctgctgattg gttcgctgac catttccggg tgcgggacgg cgttaccaga aactcagaag
     6841 gttcgtccaa ccaaaccgac tctgacggca gtttacgaga gagatgatag ggtctgcttc
     6901 agtaagccag atgctacaca attaggcttg tacatattgt cgttagaacg cggctacaat
     6961 taatacataa ccttatgtat catacacata cgatttaggt gacactatag aatacacgga
     7021 attaattc
//
