LOCUS       sequence-223910-        3279 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Multisite gateway vector for 3' tagging with rab25b.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3279)
  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 3279)
  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-rab25b
FEATURES             Location/Qualifiers
     source          1..3279
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(1..25)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(665..760)
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(797..816)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(798..816)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(821..837)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(821..837)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(939..958)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             950..1759
                     /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(1017..1036)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      1906..2494
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2395..2414
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2648..2665
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2824..2851)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(2943..3029)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3078..3100
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3092..3109
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3093..3109
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gttcaacttt cttgtacaaa gtggtggggt ctgatgaggc ctacaacttc gttttcaaag
       61 tggttcttat tggagaatcc ggtgttggca aaagcaactt gctgtcacgg ttcaccaaga
      121 acgagtttag ccacgacagc cggacgacca tcggcgtgga gttcagcact agaactgtgc
      181 agctgaacgg actcactatt aaagctcaga tctgggacac cgctggattg gagcggtacc
      241 gcgccatcac ctctgcatat tatcgaggag cggtcggagc tctgcttgtt tatgatattt
      301 ccaagcatct gacgtatgaa agtgccgagc gctggctgaa ggagctctac gatcacgcag
      361 atccacacat cgtagtgatg ctggtgggaa ataaatcaga cctggcagcc gtgcgctccg
      421 ttcctacaga ggacgccaaa gactttgcag agaagaatgg tcttctgttc atggaaacct
      481 cagctctgga gtcagtaaat gtggaagcag cgttcaacac tgtgcttaca gaaatccaca
      541 agaaggtgaa cagcaaagaa gtgacccgcg gctccatcag tgcagtcacg ttgtctcaac
      601 ctaaaactcc agcagaaacc caggaggaga agaaaacctg ctgtaaaaac ttgtgaaaga
      661 gtcacaactt tattatacaa agttggcatt ataaaaaagc attgcttatc aatttgttgc
      721 aacgaacagg tcactatcag tcaaaataaa atcattattt ggagctccat ggtagcgtta
      781 acgcggccgc gatatcccct atagtgagtc gtattacatg gtcatagctg tttcctggca
      841 gctctggccc gtgtctcaaa atctctgatg ttacattgca caagataaaa atatatcatc
      901 atgaacaata aaactgtctg cttacataaa cagtaataca aggggtgtta tgagccatat
      961 tcaacgggaa acgtcgaggc cgcgattaaa ttccaacatg gatgctgatt tatatgggta
     1021 taaatgggct cgcgataatg tcgggcaatc aggtgcgaca atctatcgct tgtatgggaa
     1081 gcccgatgcg ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca atgatgttac
     1141 agatgagatg gtcagactaa actggctgac ggaatttatg cctcttccga ccatcaagca
     1201 ttttatccgt actcctgatg atgcatggtt actcaccact gcgatccccg gaaaaacagc
     1261 attccaggta ttagaagaat atcctgattc aggtgaaaat attgttgatg cgctggcagt
     1321 gttcctgcgc cggttgcatt cgattcctgt ttgtaattgt ccttttaaca gcgatcgcgt
     1381 atttcgtctc gctcaggcgc aatcacgaat gaataacggt ttggttgatg cgagtgattt
     1441 tgatgacgag cgtaatggct ggcctgttga acaagtctgg aaagaaatgc ataaactttt
     1501 gccattctca ccggattcag tcgtcactca tggtgatttc tcacttgata accttatttt
     1561 tgacgagggg aaattaatag gttgtattga tgttggacga gtcggaatcg cagaccgata
     1621 ccaggatctt gccatcctat ggaactgcct cggtgagttt tctccttcat tacagaaacg
     1681 gctttttcaa aaatatggta ttgataatcc tgatatgaat aaattgcagt ttcatttgat
     1741 gctcgatgag tttttctaat cagaattggt taattggttg taacactggc agagcattac
     1801 gctgacttga cgggacggcg caagctcatg accaaaatcc cttaacgtga gttacgcgtc
     1861 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     1921 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     1981 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     2041 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     2101 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     2161 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     2221 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     2281 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     2341 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     2401 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     2461 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     2521 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     2581 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
     2641 cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct
     2701 ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc
     2761 gggcagtgag cgcaacgcaa ttaatacgcg taccgctagc caggaagagt ttgtagaaac
     2821 gcaaaaaggc catccgtcag gatggccttc tgcttagttt gatgcctggc agtttatggc
     2881 gggcgtcctg cccgccaccc tccgggccgt tgcttcacaa cgttcaaatc cgctcccggc
     2941 ggatttgtcc tactcaggag agcgttcacc gacaaacaac agataaaacg aaaggcccag
     3001 tcttccgact gagcctttcg ttttatttga tgcctggcag ttccctactc tcgcgttaac
     3061 gctagcatgg atgttttccc agtcacgacg ttgtaaaacg acggccagtc ttaagctcgg
     3121 gccctgcagc tctagagctc gaattctaca ggtcactaat accatctaag tagttggttc
     3181 atagtgactg catatgttgt gttttacagt attatgtagt ctgtttttta tgcaaaatct
     3241 aatttaatat attgatattt atatcatttt acgtttctc
//
