LOCUS       sequence-224065-        3280 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Multisite gateway vector for 3' tagging with dominant negative 
            rab4a.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3280)
  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 3280)
  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-rab4a-DN
FEATURES             Location/Qualifiers
     source          1..3280
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(1..125)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(766..861)
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(898..917)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(899..917)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(922..938)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(922..938)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1040..1059)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1051..1860
                     /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(1118..1137)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2007..2595
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2496..2515
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2749..2766
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2925..2952)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3044..3130)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3179..3201
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3193..3210
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3194..3210
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 catagtgact gcatatgttg tgttttacag tattatgtag tctgtttttt atgcaaaatc
       61 taatttaata tattgatatt tatatcattt tacgtttctc gttcaacttt cttgtacaaa
      121 gtggtgtcag agacatacga tttcttgttt aagtttcttg tgattgggaa tgctggaaca
      181 ggaaaaaact gtcttcttca tcagttcata gaaaaaagat tcaaagatga ctcaaatcac
      241 actattgggg ttgaatttgg ttcgaagata atcagcgtgg tcaataagtt tgtaaagctc
      301 cagatctggg acacagcagg acaggagcgt ttcaggtctg taactaggag ttattacaga
      361 ggcgcagcag gggctcttct agtctatgac atcactagtc gagaaacata caacgctcta
      421 accaactggc tgacagatgc ccggatgttg gccagtcaga acatagtgat catactatgt
      481 ggaaacaaga aggatctgga tgctgaccgc gaggtgacct ttctggaggc ttcacgcttc
      541 gctcaggaga atgagcttat gtttttggag accagtgctc tgacagggga gaatgtggag
      601 gaagctttcg tccagtgtgc tcgaaaaatc cttaacaaaa ttgaatcagg agagctggac
      661 ccagagcgga tgggttcagg aatccagtat ggagacgcgg ctcttcgaca gctgcgttca
      721 cccagacggg cacaagccga gagcatccag gagtgtggct gttaacaact ttattataca
      781 aagttggcat tataaaaaag cattgcttat caatttgttg caacgaacag gtcactatca
      841 gtcaaaataa aatcattatt tggagctcca tggtagcgtt aacgcggccg cgatatcccc
      901 tatagtgagt cgtattacat ggtcatagct gtttcctggc agctctggcc cgtgtctcaa
      961 aatctctgat gttacattgc acaagataaa aatatatcat catgaacaat aaaactgtct
     1021 gcttacataa acagtaatac aaggggtgtt atgagccata ttcaacggga aacgtcgagg
     1081 ccgcgattaa attccaacat ggatgctgat ttatatgggt ataaatgggc tcgcgataat
     1141 gtcgggcaat caggtgcgac aatctatcgc ttgtatggga agcccgatgc gccagagttg
     1201 tttctgaaac atggcaaagg tagcgttgcc aatgatgtta cagatgagat ggtcagacta
     1261 aactggctga cggaatttat gcctcttccg accatcaagc attttatccg tactcctgat
     1321 gatgcatggt tactcaccac tgcgatcccc ggaaaaacag cattccaggt attagaagaa
     1381 tatcctgatt caggtgaaaa tattgttgat gcgctggcag tgttcctgcg ccggttgcat
     1441 tcgattcctg tttgtaattg tccttttaac agcgatcgcg tatttcgtct cgctcaggcg
     1501 caatcacgaa tgaataacgg tttggttgat gcgagtgatt ttgatgacga gcgtaatggc
     1561 tggcctgttg aacaagtctg gaaagaaatg cataaacttt tgccattctc accggattca
     1621 gtcgtcactc atggtgattt ctcacttgat aaccttattt ttgacgaggg gaaattaata
     1681 ggttgtattg atgttggacg agtcggaatc gcagaccgat accaggatct tgccatccta
     1741 tggaactgcc tcggtgagtt ttctccttca ttacagaaac ggctttttca aaaatatggt
     1801 attgataatc ctgatatgaa taaattgcag tttcatttga tgctcgatga gtttttctaa
     1861 tcagaattgg ttaattggtt gtaacactgg cagagcatta cgctgacttg acgggacggc
     1921 gcaagctcat gaccaaaatc ccttaacgtg agttacgcgt cgttccactg agcgtcagac
     1981 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     2041 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     2101 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     2161 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     2221 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     2281 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     2341 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     2401 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     2461 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     2521 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     2581 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     2641 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     2701 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
     2761 agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt
     2821 cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca
     2881 attaatacgc gtaccgctag ccaggaagag tttgtagaaa cgcaaaaagg ccatccgtca
     2941 ggatggcctt ctgcttagtt tgatgcctgg cagtttatgg cgggcgtcct gcccgccacc
     3001 ctccgggccg ttgcttcaca acgttcaaat ccgctcccgg cggatttgtc ctactcagga
     3061 gagcgttcac cgacaaacaa cagataaaac gaaaggccca gtcttccgac tgagcctttc
     3121 gttttatttg atgcctggca gttccctact ctcgcgttaa cgctagcatg gatgttttcc
     3181 cagtcacgac gttgtaaaac gacggccagt cttaagctcg ggccctgcag ctctagagct
     3241 cgaattctac aggtcactaa taccatctaa gtagttggtt
//
