LOCUS       sequence-178169-        4054 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MultiSite Gateway entry clones, second fragment (attL5, attL2) Human
            YAP1 CDS.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4054)
  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 4054)
  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
FEATURES             Location/Qualifiers
     source          1..4054
                     /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             713..736
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        complement(2133..2151)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2134..2151)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     protein_bind    complement(2166..2265)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2295..2314)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2296..2314)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2319..2335)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2319..2335)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2437..2456)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2448..3257
                     /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(2515..2534)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3404..3992
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3893..3912
                     /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 agttgtaagc tcggatccac tagtaacggc cgccagtgtg ctggaattca tggactacaa
      721 ggacgacgat gacaaggatc ccgggcagca gccgccgcct caaccggccc cccagggcca
      781 agggcagccg ccttcgcagc ccccgcaggg gcagggcccg ccgtccggac ccgggcaacc
      841 ggcacccgcg gcgacccagg cggcgccgca ggcacccccc gccgggcatc agatcgtgca
      901 cgtccgcggg gactcggaga ccgacctgga ggcgctcttc aacgccgtca tgaaccccaa
      961 gacggccaac gtgccccaga ccgtgcccat gaggctccgg aagctgcccg actccttctt
     1021 caagccgccg gagcccaaat cccactcccg acaggccagt actgatgcag gcactgcagg
     1081 agccctgact ccacagcatg ttcgagctca ttcctctcca gcttctctgc agttgggagc
     1141 tgtttctcct gggacactga cccccactgg agtagtctct ggcccagcag ctacacccac
     1201 agctcagcat cttcgacagt cttcttttga gatacctgat gatgtacctc tgccagcagg
     1261 ttgggagatg gcaaagacat cttctggtca gagatacttc ttaaatcaca tcgatcagac
     1321 aacaacatgg caggacccca ggaaggccat gctgtcccag atgaacgtca cagcccccac
     1381 cagtccacca gtgcagcaga atatgatgaa ctcggcttca gccatgaacc agagaatcag
     1441 tcagagtgct ccagtgaaac agccaccacc cctggctccc cagagcccac agggaggcgt
     1501 catgggtggc agcaactcca accagcagca acagatgcga ctgcagcaac tgcagatgga
     1561 gaaggagagg ctgcggctga aacagcaaga actgcttcgg caggtgaggc cacaggagtt
     1621 agccctgcgt agccagttac caacactgga gcaggatggt gggactcaaa atccagtgtc
     1681 ttctcccggg atgtctcagg aattgagaac aatgacgacc aatagctcag atcctttcct
     1741 taacagtggc acctatcact ctcgagatga gagtacagac agtggactaa gcatgagcag
     1801 ctacagtgtc cctcgaaccc cagatgactt cctgaacagt gtggatgaga tggatacagg
     1861 tgatactatc aaccaaagca ccctgccctc acagcagaac cgtttcccag actaccttga
     1921 agccattcct gggacaaatg tggaccttgg aacactggaa ggagatggaa tgaacataga
     1981 aggagaggag ctgatgccaa gtctgcagga agctttgagt tctgacatcc ttaatgacat
     2041 ggagtctgtt ttggctgcca ccaagctaga taaagaaagc tttcttacat ggttataggc
     2101 ggccgctcga gcatgcatct agagggccct attctatagt gtcacctaaa tgctagagct
     2161 cgctaaccca gctttcttgt acaaagttgg cattataaga aagcattgct tatcaatttg
     2221 ttgcaacgaa caggtcacta tcagtcaaaa taaaatcatt atttgccatc cagctgcagg
     2281 gcggccgcga tatcccctat agtgagtcgt attacatggt catagctgtt tcctggcagc
     2341 tctggcccgt gtctcaaaat ctctgatgtt acattgcaca agataaaaat atatcatcat
     2401 gaacaataaa actgtctgct tacataaaca gtaatacaag gggtgttatg agccatattc
     2461 aacgggaaac gtcgaggccg cgattaaatt ccaacatgga tgctgattta tatgggtata
     2521 aatgggctcg cgataatgtc gggcaatcag gtgcgacaat ctatcgcttg tatgggaagc
     2581 ccgatgcgcc agagttgttt ctgaaacatg gcaaaggtag cgttgccaat gatgttacag
     2641 atgagatggt cagactaaac tggctgacgg aatttatgcc tcttccgacc atcaagcatt
     2701 ttatccgtac tcctgatgat gcatggttac tcaccactgc gatccccgga aaaacagcat
     2761 tccaggtatt agaagaatat cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt
     2821 tcctgcgccg gttgcattcg attcctgttt gtaattgtcc ttttaacagc gatcgcgtat
     2881 ttcgtctcgc tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg agtgattttg
     2941 atgacgagcg taatggctgg cctgttgaac aagtctggaa agaaatgcat aaacttttgc
     3001 cattctcacc ggattcagtc gtcactcatg gtgatttctc acttgataac cttatttttg
     3061 acgaggggaa attaataggt tgtattgatg ttggacgagt cggaatcgca gaccgatacc
     3121 aggatcttgc catcctatgg aactgcctcg gtgagttttc tccttcatta cagaaacggc
     3181 tttttcaaaa atatggtatt gataatcctg atatgaataa attgcagttt catttgatgc
     3241 tcgatgagtt tttctaatca gaattggtta attggttgta acactggcag agcattacgc
     3301 tgacttgacg ggacggcgca agctcatgac caaaatccct taacgtgagt tacgcgtcgt
     3361 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     3421 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     3481 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     3541 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     3601 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     3661 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     3721 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     3781 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     3841 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     3901 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     3961 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     4021 tcctggcctt ttgctggcct tttgctcaca tgtt
//
