LOCUS       sequence-457061-        3385 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry vector for optimized GFP-TEAD inhibitor (TEADiv2).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3385)
  AUTHORS   Branch B, Yuan Y, Cascone M, Raimondi F, Iglesias-Bartolome R
  TITLE     An improved TEAD dominant-negative protein inhibitor to study Hippo 
            YAP1/TAZ-dependent transcription.
  JOURNAL   bioRxiv [Preprint]. 2024 Oct 3:2024.10.03.615022. doi: 
            10.1101/2024.10.03.615022.
  PUBMED    39502361
REFERENCE   2  (bases 1 to 3385)
  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: "bioRxiv 
            [Preprint]. 2024 Oct 3:2024.10.03.615022. doi: 
            10.1101/2024.10.03.615022."
COMMENT     Sequence Label: pENTR-EGFP-TEADiv2
FEATURES             Location/Qualifiers
     source          1..3385
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     384..403
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      648..734
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      826..853
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     861..884
                     /label=pENTR-F
                     /note="pENTR vectors, forward primer"
     protein_bind    903..1002
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     regulatory      1062..1071
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1068..1070,1071..1073,1074..1784)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGVK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1113..1134)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1374..1393)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1721..1742
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2094..2114
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     protein_bind    complement(2159..2258)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2370..2389)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2381..3190
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(2448..2467)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      join(3280..3385,1..483)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
       61 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
      121 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
      181 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
      241 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
      301 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
      361 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
      421 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
      481 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
      541 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc tagcatggat
      601 ctcggggacg tctaactact aagcgagagt agggaactgc caggcatcaa ataaaacgaa
      661 aggctcagtc ggaagactgg gcctttcgtt ttatctgttg tttgtcggtg aacgctctcc
      721 tgagtaggac aaatccgccg ggagcggatt tgaacgttgt gaagcaacgg cccggagggt
      781 ggcgggcagg acgcccgcca taaactgcca ggcatcaaac taagcagaag gccatcctga
      841 cggatggcct ttttgcgttt ctacaaactc ttcctgttag ttagttactt aagctcgggc
      901 cccaaataat gattttattt tgactgatag tgacctgttc gttgcaacaa attgataagc
      961 aatgcttttt tataatgcca actttgtaca aaaaagcagg ctctttaaag gaaccaattc
     1021 agtcgacggc cattgtggcc atcaagcttg gtaccagatc tgccaccatg gtgagcaagg
     1081 gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg
     1141 gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc
     1201 tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc
     1261 tgacctacgg cgtgcagtgc ttcagccgct accccgacca catgaagcag cacgacttct
     1321 ttaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg
     1381 gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg
     1441 agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca
     1501 actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc gtcaaggtga
     1561 acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc
     1621 agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc
     1681 agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg
     1741 tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtccgga ctcagatctg
     1801 gatcctcagt agacgaccac tttgcgaaag cactgggtga tacatggctg caaataggtg
     1861 acccccctgt cgcgaccaat cccaaaacgg ccaatgttcc gcaaacagtg cccatgcggt
     1921 tgcggaagtt gccggagtct tttttcaaag agcccgaggg agaccctcca gtggcgacga
     1981 accccaaacc ctcttcatgg cgaaaaaaga ttctgcccga gagctttttt aaagaaccgg
     2041 gggacccgcc agttgcgaca aagcgaacag cggatggcag tgagttcgag tccccgaaga
     2101 aaaagcgcaa ggttgagtaa gcggccgcac tcgagatatc ggcctccagg gcctctagac
     2161 ccagctttct tgtacaaagt tggcattata agaaagcatt gcttatcaat ttgttgcaac
     2221 gaacaggtca ctatcagtca aaataaaatc attatttgcc atccagctgc agctctggcc
     2281 cgtgtctcaa aatctctgat gttacattgc acaagataaa aatatatcat catgaacaat
     2341 aaaactgtct gcttacataa acagtaatac aaggggtgtt atgagccata ttcaacggga
     2401 aacgtcgagg ccgcgattaa attccaacat ggatgctgat ttatatgggt ataaatgggc
     2461 tcgcgataat gtcgggcaat caggtgcgac aatctatcgc ttgtatggga agcccgatgc
     2521 gccagagttg tttctgaaac atggcaaagg tagcgttgcc aatgatgtta cagatgagat
     2581 ggtcagacta aactggctga cggaatttat gcctcttccg accatcaagc attttatccg
     2641 tactcctgat gatgcatggt tactcaccac tgcgatcccc ggaaaaacag cattccaggt
     2701 attagaagaa tatcctgatt caggtgaaaa tattgttgat gcgctggcag tgttcctgcg
     2761 ccggttgcat tcgattcctg tttgtaattg tccttttaac agcgatcgcg tatttcgtct
     2821 cgctcaggcg caatcacgaa tgaataacgg tttggttgat gcgagtgatt ttgatgacga
     2881 gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg cataaacttt tgccattctc
     2941 accggattca gtcgtcactc atggtgattt ctcacttgat aaccttattt ttgacgaggg
     3001 gaaattaata ggttgtattg atgttggacg agtcggaatc gcagaccgat accaggatct
     3061 tgccatccta tggaactgcc tcggtgagtt ttctccttca ttacagaaac ggctttttca
     3121 aaaatatggt attgataatc ctgatatgaa taaattgcag tttcatttga tgctcgatga
     3181 gtttttctaa tcagaattgg ttaattggtt gtaacattat tcagattggg ccccgttcca
     3241 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     3301 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     3361 tcaagagcta ccaactcttt ttccg
//
