LOCUS       sequence-223908-        3286 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Multisite gateway vector for 3' tagging with rab25a.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3286)
  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 3286)
  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-rab25a
FEATURES             Location/Qualifiers
     source          1..3286
                     /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(672..767)
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(804..823)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(805..823)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(828..844)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(828..844)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(946..965)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             957..1766
                     /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(1024..1043)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      1913..2501
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2402..2421
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2655..2672
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2831..2858)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(2950..3036)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3085..3107
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3099..3116
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3100..3116
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gttcaacttt cttgtacaaa gtggtgggga cagatttagc ctacaacttt gtcttcaaag
       61 tggttttaat tggagaatca ggggttggaa agagcaacct gctgtcaagg tttaccaaaa
      121 acgagttcaa ccatgacagt cgcaccacta ttggcgtgga gttcagcact cgctctgttc
      181 aggtggacag tatcaccatc aaagcccaga tctgggacac tgcgggtcta gagcgctaca
      241 gggccatcac ctcagcgtat tacagaggag cagtgggtgc gctgctggtc tatgacatca
      301 ccaaacacct gacctatgag agcgtggagc gctggctgaa ggaactgtat gatcacgctg
      361 atcctcatat cgtggtcatg ctggtgggca ataaaaatga cctggcaact gtgcgcactg
      421 tacccaccga ggaggccaaa gactttgcag atgccaatgg cttacttttt atggagactt
      481 cagcattaga gtcgactaat gttgaatctg ctttcctcga agttctaaca gcaatacacc
      541 agaaagttgc tagtaaagaa gtgaccagag gctcgatcag cgccgtgact cttggacaga
      601 cacacggtcc gtccagggaa ttccaggaga gccgaagctg ctgcaaaaac tcctgaacac
      661 accagaggac acaactttat tatacaaagt tggcattata aaaaagcatt gcttatcaat
      721 ttgttgcaac gaacaggtca ctatcagtca aaataaaatc attatttgga gctccatggt
      781 agcgttaacg cggccgcgat atcccctata gtgagtcgta ttacatggtc atagctgttt
      841 cctggcagct ctggcccgtg tctcaaaatc tctgatgtta cattgcacaa gataaaaata
      901 tatcatcatg aacaataaaa ctgtctgctt acataaacag taatacaagg ggtgttatga
      961 gccatattca acgggaaacg tcgaggccgc gattaaattc caacatggat gctgatttat
     1021 atgggtataa atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc tatcgcttgt
     1081 atgggaagcc cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc gttgccaatg
     1141 atgttacaga tgagatggtc agactaaact ggctgacgga atttatgcct cttccgacca
     1201 tcaagcattt tatccgtact cctgatgatg catggttact caccactgcg atccccggaa
     1261 aaacagcatt ccaggtatta gaagaatatc ctgattcagg tgaaaatatt gttgatgcgc
     1321 tggcagtgtt cctgcgccgg ttgcattcga ttcctgtttg taattgtcct tttaacagcg
     1381 atcgcgtatt tcgtctcgct caggcgcaat cacgaatgaa taacggtttg gttgatgcga
     1441 gtgattttga tgacgagcgt aatggctggc ctgttgaaca agtctggaaa gaaatgcata
     1501 aacttttgcc attctcaccg gattcagtcg tcactcatgg tgatttctca cttgataacc
     1561 ttatttttga cgaggggaaa ttaataggtt gtattgatgt tggacgagtc ggaatcgcag
     1621 accgatacca ggatcttgcc atcctatgga actgcctcgg tgagttttct ccttcattac
     1681 agaaacggct ttttcaaaaa tatggtattg ataatcctga tatgaataaa ttgcagtttc
     1741 atttgatgct cgatgagttt ttctaatcag aattggttaa ttggttgtaa cactggcaga
     1801 gcattacgct gacttgacgg gacggcgcaa gctcatgacc aaaatccctt aacgtgagtt
     1861 acgcgtcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     1921 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     1981 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     2041 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     2101 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     2161 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     2221 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     2281 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     2341 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     2401 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     2461 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     2521 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     2581 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     2641 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac
     2701 cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact
     2761 ggaaagcggg cagtgagcgc aacgcaatta atacgcgtac cgctagccag gaagagtttg
     2821 tagaaacgca aaaaggccat ccgtcaggat ggccttctgc ttagtttgat gcctggcagt
     2881 ttatggcggg cgtcctgccc gccaccctcc gggccgttgc ttcacaacgt tcaaatccgc
     2941 tcccggcgga tttgtcctac tcaggagagc gttcaccgac aaacaacaga taaaacgaaa
     3001 ggcccagtct tccgactgag cctttcgttt tatttgatgc ctggcagttc cctactctcg
     3061 cgttaacgct agcatggatg ttttcccagt cacgacgttg taaaacgacg gccagtctta
     3121 agctcgggcc ctgcagctct agagctcgaa ttctacaggt cactaatacc atctaagtag
     3181 ttggttcata gtgactgcat atgttgtgtt ttacagtatt atgtagtctg ttttttatgc
     3241 aaaatctaat ttaatatatt gatatttata tcattttacg tttctc
//
