LOCUS       sequence-178164-        3940 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MultiSite Gateway entry clones, second fragment (attL5, attL2) Human
            TAZ S89A mutant CDS.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3940)
  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 3940)
  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-TAZ CA
FEATURES             Location/Qualifiers
     source          1..3940
                     /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             698..763
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     protein_bind    complement(2052..2151)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2181..2200)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2182..2200)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2205..2221)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2205..2221)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2323..2342)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2334..3143
                     /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(2401..2420)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3290..3878
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3779..3798
                     /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 agttgtagtg aaccgtcaga attgatctgg taccatggac tacaaagacc atgacggtga
      721 ttataaagat catgacatcg attacaagga tgacgatgac aagcttggga atccggcctc
      781 ggcgccccct ccgctcccgc cgcctgggca gcaagtgatc cacgtcacgc aggacctaga
      841 cacagacctc gaagccctct tcaactctgt catgaatccg aagcctagct cgtggcggaa
      901 gaagatcctg ccggagtctt tctttaagga gcctgattcg ggctcgcact cgcgccagtc
      961 cagcaccgac tcgtcgggcg gccacccggg gcctcgactg gctgggggtg cccagcatgt
     1021 ccgctcgcac gcctcgccag cgtccctgca gctgggcacc ggcgcgggtg ctgcgggtag
     1081 ccccgcgcag cagcacgcgc acctccgcca gcagtcctac gacgtgaccg acgagctgcc
     1141 actgcccccg ggctgggaga tgaccttcac ggccactggc cagaggtact tcctcaatca
     1201 catagaaaaa atcaccacat ggcaagaccc taggaaggcg atgaatcagc ctctgaatca
     1261 tatgaacctc caccctgccg tcagttccac accagtgcct cagaggtcca tggcagtatc
     1321 ccagccaaat ctcgtgatga atcaccaaca ccagcagcag atggccccca gtaccctgag
     1381 ccagcagaac caccccactc agaacccacc cgcagggctc atgagtatgc ccaatgcgct
     1441 gaccactcag cagcagcagc agcagaaact gcggcttcag agaatccaga tggagagaga
     1501 aaggattcga atgcgccaag aggagctcat gaggcaggaa gctgccctct gtcgacagct
     1561 ccccatggaa gctgagactc ttgccccagt tcaggctgct gtcaacccac ccacgatgac
     1621 cccagacatg agatccatca ctaataatag ctcagatcct ttcctcaatg gagggccata
     1681 tcattcgagg gagcagagca ctgacagtgg cctggggtta gggtgctaca gtgtccccac
     1741 aactccggag gacttcctca gcaatgtgga tgagatggat acaggagaaa acgcaggaca
     1801 aacacccatg aacatcaatc cccaacagac ccgtttccct gatttccttg actgtcttcc
     1861 aggaacaaac gttgacttag gaactttgga atctgaagac ctgatccccc tcttcaatga
     1921 tgtagagtct gctctgaaca aaagtgagcc ctttctaacc tggctgtaag aattcaaggg
     1981 cgatcccggg tggcatccct gtgacccctc cccagtgcct ctcctggccc tggaagttgc
     2041 cactccagtg aacccagctt tcttgtacaa agttggcatt ataagaaagc attgcttatc
     2101 aatttgttgc aacgaacagg tcactatcag tcaaaataaa atcattattt gccatccagc
     2161 tgcagggcgg ccgcgatatc ccctatagtg agtcgtatta catggtcata gctgtttcct
     2221 ggcagctctg gcccgtgtct caaaatctct gatgttacat tgcacaagat aaaaatatat
     2281 catcatgaac aataaaactg tctgcttaca taaacagtaa tacaaggggt gttatgagcc
     2341 atattcaacg ggaaacgtcg aggccgcgat taaattccaa catggatgct gatttatatg
     2401 ggtataaatg ggctcgcgat aatgtcgggc aatcaggtgc gacaatctat cgcttgtatg
     2461 ggaagcccga tgcgccagag ttgtttctga aacatggcaa aggtagcgtt gccaatgatg
     2521 ttacagatga gatggtcaga ctaaactggc tgacggaatt tatgcctctt ccgaccatca
     2581 agcattttat ccgtactcct gatgatgcat ggttactcac cactgcgatc cccggaaaaa
     2641 cagcattcca ggtattagaa gaatatcctg attcaggtga aaatattgtt gatgcgctgg
     2701 cagtgttcct gcgccggttg cattcgattc ctgtttgtaa ttgtcctttt aacagcgatc
     2761 gcgtatttcg tctcgctcag gcgcaatcac gaatgaataa cggtttggtt gatgcgagtg
     2821 attttgatga cgagcgtaat ggctggcctg ttgaacaagt ctggaaagaa atgcataaac
     2881 ttttgccatt ctcaccggat tcagtcgtca ctcatggtga tttctcactt gataacctta
     2941 tttttgacga ggggaaatta ataggttgta ttgatgttgg acgagtcgga atcgcagacc
     3001 gataccagga tcttgccatc ctatggaact gcctcggtga gttttctcct tcattacaga
     3061 aacggctttt tcaaaaatat ggtattgata atcctgatat gaataaattg cagtttcatt
     3121 tgatgctcga tgagtttttc taatcagaat tggttaattg gttgtaacac tggcagagca
     3181 ttacgctgac ttgacgggac ggcgcaagct catgaccaaa atcccttaac gtgagttacg
     3241 cgtcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     3301 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     3361 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     3421 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     3481 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     3541 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     3601 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     3661 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     3721 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     3781 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     3841 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     3901 tacggttcct ggccttttgc tggccttttg ctcacatgtt
//
