LOCUS       sequence-178078-        4132 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MultiSite Gateway entry clones, second fragment (attL5, attL2) Human
            YAP1 S127A mutant CDS.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4132)
  AUTHORS   Jackel C, Nogueira MS, Ehni N, Kraus C, Ranke J, Dohmann M, Noessner
            E, Nelson PJ
  TITLE     A vector platform for the rapid and efficient engineering of stable 
            complex transgenes.
  JOURNAL   Sci Rep. 2016 Oct 3;6:34365. doi: 10.1038/srep34365.
  PUBMED    27694838
REFERENCE   2  (bases 1 to 4132)
  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: "Sci Rep."; 
            date: "2016-10-3"; volume: "6:34365. doi"; pages: " 
            10.1038/srep34365"
COMMENT     Sequence Label: pENTR221-YAP1 CA
FEATURES             Location/Qualifiers
     source          1..4132
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     536..553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    570..665
                     /gene="mutant version of attL"
                     /label=attL5
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             682..753
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags"
                     /label=3xFLAG
                     /translation="DYKDDDDKDYKDDDDKDYKDDDDK"
     protein_bind    complement(2244..2343)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2373..2392)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2374..2392)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2397..2413)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2397..2413)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2515..2534)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2526..3335
                     /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(2593..2612)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3482..4070
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3971..3990
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac
      601 ctgttcgttg caacaaattg atgagcaatg cttttttata atgccaactt tgtatacaaa
      661 agttgtacag tcgacaccat ggactacaag gacgacgatg acaaggatta caaagatgac
      721 gacgataagg actataagga cgatgatgat aaactggtac ctatggatcc cgggcagcag
      781 ccgccgcctc aaccggcccc ccagggccaa gggcagccgc cttcgcagcc cccgcagggg
      841 cagggcccgc cgtccggacc cgggcaaccg gcacccgcgg cgacccaggc ggcgccgcag
      901 gcaccccccg ccgggcatca gatcgtgcac gtccgcgggg actcggagac cgacctggag
      961 gcgctcttca acgccgtcat gaaccccaag acggccaacg tgccccagac cgtgcccatg
     1021 aggctccgga agctgcccga ctccttcttc aagccgccgg agcccaaatc ccactcccga
     1081 caggccagta ctgatgcagg cactgcagga gccctgactc cacagcatgt tcgagctcat
     1141 gcctctccag cttctctgca gttgggagct gtttctcctg ggacactgac ccccactgga
     1201 gtagtctctg gcccagcagc tacacccaca gctcagcatc ttcgacagtc ttcttttgag
     1261 atacctgatg atgtacctct gccagcaggt tgggagatgg caaagacatc ttctggtcag
     1321 agatacttct taaatcacat cgatcagaca acaacatggc aggaccccag gaaggccatg
     1381 ctgtcccaga tgaacgtcac agcccccacc agtccaccag tgcagcagaa tatgatgaac
     1441 tcggcttcag gtcctcttcc tgatggatgg gaacaagcca tgactcagga tggagaaatt
     1501 tactatataa accataagaa caagaccacc tcttggctag acccaaggct tgaccctcgt
     1561 tttgccatga accagagaat cagtcagagt gctccagtga aacagccacc acccctggct
     1621 ccccagagcc cacagggagg cgtcatgggt ggcagcaact ccaaccagca gcaacagatg
     1681 cgactgcagc aactgcagat ggagaaggag aggctgcggc tgaaacagca agaactgctt
     1741 cggcaggagt tagccctgcg tagccagtta ccaacactgg agcaggatgg tgggactcaa
     1801 aatccagtgt cttctcccgg gatgtctcag gaattgagaa caatgacgac caatagctca
     1861 gatcctttcc ttaacagtgg cacctatcac tctcgagatg agagtacaga cagtggacta
     1921 agcatgagca gctacagtgt ccctcgaacc ccagatgact tcctgaacag tgtggatgag
     1981 atggatacag gtgatactat caaccaaagc accctgccct cacagcagaa ccgtttccca
     2041 gactaccttg aagccattcc tgggacaaat gtggaccttg gaacactgga aggagatgga
     2101 atgaacatag aaggagagga gctgatgcca agtctgcagg aagctttgag ttctgacatc
     2161 cttaatgaca tggagtctgt tttggctgcc accaagctag ataaagaaag ctttcttaca
     2221 tggttataga gccctcaggc agaacccagc tttcttgtac aaagttggca ttataagaaa
     2281 gcattgctta tcaatttgtt gcaacgaaca ggtcactatc agtcaaaata aaatcattat
     2341 ttgccatcca gctgcagggc ggccgcgata tcccctatag tgagtcgtat tacatggtca
     2401 tagctgtttc ctggcagctc tggcccgtgt ctcaaaatct ctgatgttac attgcacaag
     2461 ataaaaatat atcatcatga acaataaaac tgtctgctta cataaacagt aatacaaggg
     2521 gtgttatgag ccatattcaa cgggaaacgt cgaggccgcg attaaattcc aacatggatg
     2581 ctgatttata tgggtataaa tgggctcgcg ataatgtcgg gcaatcaggt gcgacaatct
     2641 atcgcttgta tgggaagccc gatgcgccag agttgtttct gaaacatggc aaaggtagcg
     2701 ttgccaatga tgttacagat gagatggtca gactaaactg gctgacggaa tttatgcctc
     2761 ttccgaccat caagcatttt atccgtactc ctgatgatgc atggttactc accactgcga
     2821 tccccggaaa aacagcattc caggtattag aagaatatcc tgattcaggt gaaaatattg
     2881 ttgatgcgct ggcagtgttc ctgcgccggt tgcattcgat tcctgtttgt aattgtcctt
     2941 ttaacagcga tcgcgtattt cgtctcgctc aggcgcaatc acgaatgaat aacggtttgg
     3001 ttgatgcgag tgattttgat gacgagcgta atggctggcc tgttgaacaa gtctggaaag
     3061 aaatgcataa acttttgcca ttctcaccgg attcagtcgt cactcatggt gatttctcac
     3121 ttgataacct tatttttgac gaggggaaat taataggttg tattgatgtt ggacgagtcg
     3181 gaatcgcaga ccgataccag gatcttgcca tcctatggaa ctgcctcggt gagttttctc
     3241 cttcattaca gaaacggctt tttcaaaaat atggtattga taatcctgat atgaataaat
     3301 tgcagtttca tttgatgctc gatgagtttt tctaatcaga attggttaat tggttgtaac
     3361 actggcagag cattacgctg acttgacggg acggcgcaag ctcatgacca aaatccctta
     3421 acgtgagtta cgcgtcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     3481 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     3541 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     3601 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     3661 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     3721 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     3781 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     3841 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     3901 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     3961 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     4021 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     4081 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg tt
//
