LOCUS       sequence-222115-        3214 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Multisite gateway vector for 3' tagging with dominant negative 
            CDC42.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3214)
  AUTHORS   Hall TE, Martel N, Ariotti N, Xiong Z, Lo HP, Ferguson C, Rae J, Lim
            YW, Parton RG
  TITLE     In vivo cell biological screening identifies an endocytic capture 
            mechanism for T-tubule formation.
  JOURNAL   Nat Commun. 2020 Jul 24;11(1):3711. doi: 10.1038/s41467-020-17486-w.
  PUBMED    32709891
REFERENCE   2  (bases 1 to 3214)
  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: "Nat 
            Commun."; date: "2020-07-24"; volume: "11(1):3711. doi"; pages: " 
            10.1038/s41467-020-17486-w"
COMMENT     Sequence Label: p3E-CDC42-DN
FEATURES             Location/Qualifiers
     source          1..3214
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(600..695)
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(732..751)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(733..751)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(756..772)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(756..772)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(874..893)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             885..1694
                     /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(952..971)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      1841..2429
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2330..2349
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2583..2600
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2759..2786)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(2878..2964)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3013..3035
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3027..3044
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3028..3044
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(join(3115..3214,1..24))
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
ORIGIN
        1 gttcaacttt cttgtacaaa gtggctcaga cgatcaagtg cgtcgttgtt ggtgatggtg
       61 cagtgggtaa aaactgtcta ttaatctcct atacaactaa taaattcccc tccgaatatg
      121 taccaacggt ctttgataac tatgcagtta cggttatgat tggtggtgag ccgtacaccc
      181 tggggctgtt tgatactgca ggtcaggaag attacgatag attacgaccc ctgagttacc
      241 ctcagacaga tgtcttctta gtctgtttct cagttgtttc gccttcttca tttgaaaacg
      301 tcaaagaaaa gtgggtaccc gagatcaccc accactgtcc aaagacccct ttcctgctgg
      361 tggggacgca gattgatctg agagatgatc cttctactat cgaaaagctt gccaagaaca
      421 aacagaaacc catcactcca gagacggcag agaagctggc ccgcgatctg aaggctgtca
      481 aatacgtgga atgctccgtt ctgacgcaga aaggcctaaa gaacgtattt gatgaggcga
      541 ttttggcagc cctggagccc ccagagccca agaagaagcg taaatgtgtg ctgctatgac
      601 aactttatta tacaaagttg gcattataaa aaagcattgc ttatcaattt gttgcaacga
      661 acaggtcact atcagtcaaa ataaaatcat tatttggagc tccatggtag cgttaacgcg
      721 gccgcgatat cccctatagt gagtcgtatt acatggtcat agctgtttcc tggcagctct
      781 ggcccgtgtc tcaaaatctc tgatgttaca ttgcacaaga taaaaatata tcatcatgaa
      841 caataaaact gtctgcttac ataaacagta atacaagggg tgttatgagc catattcaac
      901 gggaaacgtc gaggccgcga ttaaattcca acatggatgc tgatttatat gggtataaat
      961 gggctcgcga taatgtcggg caatcaggtg cgacaatcta tcgcttgtat gggaagcccg
     1021 atgcgccaga gttgtttctg aaacatggca aaggtagcgt tgccaatgat gttacagatg
     1081 agatggtcag actaaactgg ctgacggaat ttatgcctct tccgaccatc aagcatttta
     1141 tccgtactcc tgatgatgca tggttactca ccactgcgat ccccggaaaa acagcattcc
     1201 aggtattaga agaatatcct gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc
     1261 tgcgccggtt gcattcgatt cctgtttgta attgtccttt taacagcgat cgcgtatttc
     1321 gtctcgctca ggcgcaatca cgaatgaata acggtttggt tgatgcgagt gattttgatg
     1381 acgagcgtaa tggctggcct gttgaacaag tctggaaaga aatgcataaa cttttgccat
     1441 tctcaccgga ttcagtcgtc actcatggtg atttctcact tgataacctt atttttgacg
     1501 aggggaaatt aataggttgt attgatgttg gacgagtcgg aatcgcagac cgataccagg
     1561 atcttgccat cctatggaac tgcctcggtg agttttctcc ttcattacag aaacggcttt
     1621 ttcaaaaata tggtattgat aatcctgata tgaataaatt gcagtttcat ttgatgctcg
     1681 atgagttttt ctaatcagaa ttggttaatt ggttgtaaca ctggcagagc attacgctga
     1741 cttgacggga cggcgcaagc tcatgaccaa aatcccttaa cgtgagttac gcgtcgttcc
     1801 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     1861 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     1921 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     1981 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     2041 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     2101 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     2161 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     2221 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     2281 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     2341 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     2401 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     2461 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     2521 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
     2581 gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg
     2641 cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca
     2701 gtgagcgcaa cgcaattaat acgcgtaccg ctagccagga agagtttgta gaaacgcaaa
     2761 aaggccatcc gtcaggatgg ccttctgctt agtttgatgc ctggcagttt atggcgggcg
     2821 tcctgcccgc caccctccgg gccgttgctt cacaacgttc aaatccgctc ccggcggatt
     2881 tgtcctactc aggagagcgt tcaccgacaa acaacagata aaacgaaagg cccagtcttc
     2941 cgactgagcc tttcgtttta tttgatgcct ggcagttccc tactctcgcg ttaacgctag
     3001 catggatgtt ttcccagtca cgacgttgta aaacgacggc cagtcttaag ctcgggccct
     3061 gcagctctag agctcgaatt ctacaggtca ctaataccat ctaagtagtt ggttcatagt
     3121 gactgcatat gttgtgtttt acagtattat gtagtctgtt ttttatgcaa aatctaattt
     3181 aatatattga tatttatatc attttacgtt tctc
//
