LOCUS       sequence-224025-        3309 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Multisite gateway vector for 3' tagging with rab5aa.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3309)
  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 3309)
  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-rab5aa
FEATURES             Location/Qualifiers
     source          1..3309
                     /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(795..890)
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(927..946)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(928..946)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(951..967)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(951..967)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1069..1088)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1080..1889
                     /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(1147..1166)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2036..2624
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2525..2544
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2778..2795
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2954..2981)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3073..3159)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3208..3230
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3222..3239
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3223..3239
                     /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 gtggtggcca ataggggagg agcaacacgc cccaacgggt ccaacgcggg caataagatc
      181 tgccagttta aactggtctt gctaggagag tcagcggtgg ggaagtccag cctcgttctg
      241 cgcttcgtca aagggcagtt tcacgagttt caggaaagca caataggagc ggcctttctt
      301 acacagacac tgtgcttgga tgacacgacg gtaaagtttg agatctggga cacagctgga
      361 caggagcgct accacagtct ggcccccatg tactacagag gtgcccaggc cgccattgta
      421 gtttatgaca tcaccaatga ggagtcattt gcaagagcaa agaactgggt taaagagctt
      481 cagaggcagg ctagcccaaa cattgtcatc gctctgtctg ggaacaaggc tgaccttgcc
      541 aacaagagag ctgtggactt ccaggatgct cagtcttatg cagatgacaa cagtttgctg
      601 ttcatggaga cgtcagcaaa aacctccatg aacgtgaacg agatcttcat ggccattgcg
      661 aaaaagttgc ctaagagtga gccccaagct gccggagcca acagcgggcg gagtaggggt
      721 gtggacctca cagagacagc ccaacccact aaagccccct gctgcagcaa ctaacgcaaa
      781 acaaaaatcc ccgtcaactt tattatacaa agttggcatt ataaaaaagc attgcttatc
      841 aatttgttgc aacgaacagg tcactatcag tcaaaataaa atcattattt ggagctccat
      901 ggtagcgtta acgcggccgc gatatcccct atagtgagtc gtattacatg gtcatagctg
      961 tttcctggca gctctggccc gtgtctcaaa atctctgatg ttacattgca caagataaaa
     1021 atatatcatc atgaacaata aaactgtctg cttacataaa cagtaataca aggggtgtta
     1081 tgagccatat tcaacgggaa acgtcgaggc cgcgattaaa ttccaacatg gatgctgatt
     1141 tatatgggta taaatgggct cgcgataatg tcgggcaatc aggtgcgaca atctatcgct
     1201 tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca
     1261 atgatgttac agatgagatg gtcagactaa actggctgac ggaatttatg cctcttccga
     1321 ccatcaagca ttttatccgt actcctgatg atgcatggtt actcaccact gcgatccccg
     1381 gaaaaacagc attccaggta ttagaagaat atcctgattc aggtgaaaat attgttgatg
     1441 cgctggcagt gttcctgcgc cggttgcatt cgattcctgt ttgtaattgt ccttttaaca
     1501 gcgatcgcgt atttcgtctc gctcaggcgc aatcacgaat gaataacggt ttggttgatg
     1561 cgagtgattt tgatgacgag cgtaatggct ggcctgttga acaagtctgg aaagaaatgc
     1621 ataaactttt gccattctca ccggattcag tcgtcactca tggtgatttc tcacttgata
     1681 accttatttt tgacgagggg aaattaatag gttgtattga tgttggacga gtcggaatcg
     1741 cagaccgata ccaggatctt gccatcctat ggaactgcct cggtgagttt tctccttcat
     1801 tacagaaacg gctttttcaa aaatatggta ttgataatcc tgatatgaat aaattgcagt
     1861 ttcatttgat gctcgatgag tttttctaat cagaattggt taattggttg taacactggc
     1921 agagcattac gctgacttga cgggacggcg caagctcatg accaaaatcc cttaacgtga
     1981 gttacgcgtc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     2041 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     2101 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     2161 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     2221 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     2281 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     2341 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     2401 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     2461 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     2521 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     2581 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     2641 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     2701 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     2761 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca
     2821 aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg
     2881 actggaaagc gggcagtgag cgcaacgcaa ttaatacgcg taccgctagc caggaagagt
     2941 ttgtagaaac gcaaaaaggc catccgtcag gatggccttc tgcttagttt gatgcctggc
     3001 agtttatggc gggcgtcctg cccgccaccc tccgggccgt tgcttcacaa cgttcaaatc
     3061 cgctcccggc ggatttgtcc tactcaggag agcgttcacc gacaaacaac agataaaacg
     3121 aaaggcccag tcttccgact gagcctttcg ttttatttga tgcctggcag ttccctactc
     3181 tcgcgttaac gctagcatgg atgttttccc agtcacgacg ttgtaaaacg acggccagtc
     3241 ttaagctcgg gccctgcagc tctagagctc gaattctaca ggtcactaat accatctaag
     3301 tagttggtt
//
