LOCUS       sequence-340590-        7137 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  firefly luciferase reporter vector with 18x clustered TEAD binding 
            sites.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7137)
  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 7137)
  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_18xTEAD_luc2
FEATURES             Location/Qualifiers
     source          1..7137
                     /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(636..642,643..644,645..651)
                     /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(668..674,675..676,677..683)
                     /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(700..706,707..708,709..715)
                     /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(753..759,760..761,762..768)
                     /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(785..791,792..793,794..800)
                     /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(817..823,824..825,826..832)
                     /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(839..859)
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     protein_bind    1000..1020
                     /gene="mutant version of attB"
                     /label=attB3
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     promoter        1047..1065
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1128..1147)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1129..1147)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    complement(1171..1195)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      1274..1283
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1280..2932
                     /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    2960..3548
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3013..3033)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3431..3442)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3575..3592)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3581..3805
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        complement(4015..4344)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4029..4164
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4329..4349
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(4590..5178)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4670..4689)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5349..6140,6141..6209))
                     /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     5972..5991
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6210..6314)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6382..6400
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6438..6460)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6560..6579
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6793..6812
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        7102..7120
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     7102..7119
                     /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 gcaggaagct tgcagtggaa tgtgtggaat gtgtctcaag
      661 cttgcagtgg aatgtgtgga atgtgtctca agcttgcagt ggaatgtgtg gaatgtgtct
      721 cggcctcctg caggaggcgg ccaagcttgc agtggaatgt gtggaatgtg tctcaagctt
      781 gcagtggaat gtgtggaatg tgtctcaagc ttgcagtgga atgtgtggaa tgtgtctcca
      841 acttttctat acaaagttgg caagtgcagg tgccagaaca tttctctggc ctaactggaa
      901 gacgctagcg gggggctata aaagggggtg ggggcgttcg tcctcactct agatctgcga
      961 tctaagtaag cttggcaatc cggtactgtt ggtaaagccc aactttgtat aataaagttg
     1021 agctagttgc taagtctgcc gagtagaatt aaccctcact aaagggtagg tgacactatt
     1081 ctattacatc aatcatctaa gtgcgtcttc tattacttac ctttaatccc tatagtgagt
     1141 cgtattagta gacgcgagtc aatgcatgta cgacagcttt cttgtacaaa gtggtgatat
     1201 cgaattcctg cagcccgggg gatccgcccg ggctagagcg gccgccaccg cggtggagct
     1261 cgcgactagt gacgccacca tggaagatgc caaaaacatt aagaagggcc cagcgccatt
     1321 ctacccactc gaagacggga ccgccggcga gcagctgcac aaagccatga agcgctacgc
     1381 cctggtgcct ggcaccatcg cctttaccga cgcacatatc gaggtggaca ttacctacgc
     1441 cgagtacttc gagatgagcg ttcggctggc agaagctatg aagcgctatg ggctgaatac
     1501 aaaccatcgg atcgtggtgt gcagcgagaa tagcttgcag ttcttcatgc ccgtgttggg
     1561 tgccctgttc atcggtgtgg ctgtggcccc agctaacgac atctacaacg agcgcgagct
     1621 gctgaacagc atgggcatca gccagcccac cgtcgtattc gtgagcaaga aagggctgca
     1681 aaagatcctc aacgtgcaaa agaagctacc gatcatacaa aagatcatca tcatggatag
     1741 caagaccgac taccagggct tccaaagcat gtacaccttc gtgacttccc atttgccacc
     1801 cggcttcaac gagtacgact tcgtgcccga gagcttcgac cgggacaaaa ccatcgccct
     1861 gatcatgaac agtagtggca gtaccggatt gcccaagggc gtagccctac cgcaccgcac
     1921 cgcttgtgtc cgattcagtc atgcccgcga ccccatcttc ggcaaccaga tcatccccga
     1981 caccgctatc ctcagcgtgg tgccatttca ccacggcttc ggcatgttca ccacgctggg
     2041 ctacttgatc tgcggctttc gggtcgtgct catgtaccgc ttcgaggagg agctattctt
     2101 gcgcagcttg caagactata agattcaatc tgccctgctg gtgcccacac tatttagctt
     2161 cttcgctaag agcactctca tcgacaagta cgacctaagc aacttgcacg agatcgccag
     2221 cggcggggcg ccgctcagca aggaggtagg tgaggccgtg gccaaacgct tccacctacc
     2281 aggcatccgc cagggctacg gcctgacaga aacaaccagc gccattctga tcacccccga
     2341 aggggacgac aagcctggcg cagtaggcaa ggtggtgccc ttcttcgagg ctaaggtggt
     2401 ggacttggac accggtaaga cactgggtgt gaaccagcgc ggcgagctgt gcgtccgtgg
     2461 ccccatgatc atgagcggct acgttaacaa ccccgaggct acaaacgctc tcatcgacaa
     2521 ggacggctgg ctgcacagcg gcgacatcgc ctactgggac gaggacgagc acttcttcat
     2581 cgtggaccgg ctgaagagcc tgatcaaata caagggctac caggtagccc cagccgaact
     2641 ggagagcatc ctgctgcaac accccaacat cttcgacgcc ggggtcgccg gcctgcccga
     2701 cgacgatgcc ggcgagctgc ccgccgcagt cgtcgtgctg gaacacggta aaaccataac
     2761 cgagaaggag atcgtggact atgtggccag ccaggctaca accgccaaga agctgcgcgg
     2821 tggtgttgtg ttcgtggacg aggtgcctaa aggactgacc ggcaagttgg acgcccgcaa
     2881 gatccgcgag attctcatta aggccaagaa gggcggcaag atcgccgtgt aattcgaatt
     2941 cgatatcaag cttatcgata atcaacctct ggattacaaa atttgtgaaa gattgactgg
     3001 tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta
     3061 tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct
     3121 gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt
     3181 tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac
     3241 tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg
     3301 ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaaatcatc
     3361 gtcctttcct tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg
     3421 ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct
     3481 gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc
     3541 ctccccgcat cgataccgtc gactcgctga tcagcctcga ctgtgccttc tagttgccag
     3601 ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact
     3661 gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt
     3721 ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat
     3781 gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctcgact
     3841 agagcatggc tacgtagata agtagcatgg cgggttaatc attaactaca aggaacccct
     3901 agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc
     3961 aaaggtcgcc cgacgcccgg gcggcctcag tgagcgagcg agcgcgcaga gctttttgca
     4021 aaagcctagg cctccaaaaa agcctcctca ctacttctgg aatagctcag aggccgaggc
     4081 ggcctcggcc tctgcataaa taaaaaaaat tagtcagcca tggggcggag aatgggcgga
     4141 actgggcgga gttaggggcg ggatgggcgg agttaggggc gggactatgg ttgctgacta
     4201 attgagatgc atgctttgca tacttctgcc tgctggggag cctggggact ttccacacct
     4261 ggttgctgac taattgagat gcatgctttg catacttctg cctgctgggg agcctgggga
     4321 ctttccacac cctaactgac acacattcca cagctgcatt aatgaatcgg ccaacgcgcg
     4381 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     4441 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     4501 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     4561 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     4621 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     4681 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     4741 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     4801 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     4861 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     4921 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     4981 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     5041 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     5101 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     5161 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     5221 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     5281 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     5341 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     5401 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     5461 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     5521 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     5581 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     5641 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     5701 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     5761 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     5821 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     5881 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     5941 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     6001 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     6061 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     6121 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     6181 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     6241 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     6301 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt
     6361 atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc
     6421 ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg
     6481 taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt
     6541 cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca ccattcgacg
     6601 ctctccctta tgcgactcct gcattaggaa gcagcccagt agtaggttga ggccgttgag
     6661 caccgccgcc gcaaggaatg gtgcatgcaa ggagatggcg cccaacagtc ccccggccac
     6721 ggggcctgcc accataccca cgccgaaaca agcgctcatg agcccgaagt ggcgagcccg
     6781 atcttcccca tcggtgatgt cggcgatata ggcgccagca accgcacctg tggcgccggt
     6841 gatgccggcc acgatgcgtc cggcgtagag gatctggcta gcgatgaccc tgctgattgg
     6901 ttcgctgacc atttccgggt gcgggacggc gttaccagaa actcagaagg ttcgtccaac
     6961 caaaccgact ctgacggcag tttacgagag agatgatagg gtctgcttca gtaagccaga
     7021 tgctacacaa ttaggcttgt acatattgtc gttagaacgc ggctacaatt aatacataac
     7081 cttatgtatc atacacatac gatttaggtg acactataga atacacggaa ttaattc
//
