LOCUS       sequence-224028-        3286 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Multisite gateway vector for 3' tagging with rab5b.
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-rab5b
FEATURES             Location/Qualifiers
     source          1..3286
                     /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(772..867)
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(904..923)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(905..923)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(928..944)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(928..944)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1046..1065)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1057..1866
                     /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(1124..1143)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2013..2601
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2502..2521
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2755..2772
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2931..2958)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3050..3136)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3185..3207
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3199..3216
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3200..3216
                     /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 gtggtgagta cccgtgggac aggcagggcc aatgggaacc taccgcagac caagatctgc
      181 cagtttaaac tggtgctgct gggagacatg gccgtcggca agtccagcct ggtgctgcgc
      241 tttgtgaagg gacagtttga tgagtttcag gagaccacca ttggagctgc attcttggcc
      301 cagtctgttt gtctggacga cactacagtg aagtttgaga tttgggacac agccggacaa
      361 gagcgctacc acagcttggc tcccatgtac taccgtggag cccaggcagc cattgtagtg
      421 tttgatatca ctaaacctga aacatttgag cgagcaaaag catgggtgaa agagctgcag
      481 cggcaggcca gtcccaacat agtcattgct cttgcaggaa acaaagccga tctagcagac
      541 aagagacttg tggagtatga ggaagcccag acatatgcag aagacacagg tttactgttc
      601 atggaaacct ctgccaagac tgcaatgaat gtaaacgagt tgttcctggc cattgcaaaa
      661 aagatgccaa aaacagacac ccaaaaccct acgcatgctg cgcgacacag gggcgtgaac
      721 ctgcaggacc cagacgcaca gtccactcga agctgctgtg ggaactaaag gcaactttat
      781 tatacaaagt tggcattata aaaaagcatt gcttatcaat ttgttgcaac gaacaggtca
      841 ctatcagtca aaataaaatc attatttgga gctccatggt agcgttaacg cggccgcgat
      901 atcccctata gtgagtcgta ttacatggtc atagctgttt cctggcagct ctggcccgtg
      961 tctcaaaatc tctgatgtta cattgcacaa gataaaaata tatcatcatg aacaataaaa
     1021 ctgtctgctt acataaacag taatacaagg ggtgttatga gccatattca acgggaaacg
     1081 tcgaggccgc gattaaattc caacatggat gctgatttat atgggtataa atgggctcgc
     1141 gataatgtcg ggcaatcagg tgcgacaatc tatcgcttgt atgggaagcc cgatgcgcca
     1201 gagttgtttc tgaaacatgg caaaggtagc gttgccaatg atgttacaga tgagatggtc
     1261 agactaaact ggctgacgga atttatgcct cttccgacca tcaagcattt tatccgtact
     1321 cctgatgatg catggttact caccactgcg atccccggaa aaacagcatt ccaggtatta
     1381 gaagaatatc ctgattcagg tgaaaatatt gttgatgcgc tggcagtgtt cctgcgccgg
     1441 ttgcattcga ttcctgtttg taattgtcct tttaacagcg atcgcgtatt tcgtctcgct
     1501 caggcgcaat cacgaatgaa taacggtttg gttgatgcga gtgattttga tgacgagcgt
     1561 aatggctggc ctgttgaaca agtctggaaa gaaatgcata aacttttgcc attctcaccg
     1621 gattcagtcg tcactcatgg tgatttctca cttgataacc ttatttttga cgaggggaaa
     1681 ttaataggtt gtattgatgt tggacgagtc ggaatcgcag accgatacca ggatcttgcc
     1741 atcctatgga actgcctcgg tgagttttct ccttcattac agaaacggct ttttcaaaaa
     1801 tatggtattg ataatcctga tatgaataaa ttgcagtttc atttgatgct cgatgagttt
     1861 ttctaatcag aattggttaa ttggttgtaa cactggcaga gcattacgct gacttgacgg
     1921 gacggcgcaa gctcatgacc aaaatccctt aacgtgagtt acgcgtcgtt ccactgagcg
     1981 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     2041 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     2101 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     2161 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     2221 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     2281 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     2341 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     2401 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     2461 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     2521 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     2581 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     2641 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     2701 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     2761 tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg
     2821 ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc
     2881 aacgcaatta atacgcgtac cgctagccag gaagagtttg tagaaacgca aaaaggccat
     2941 ccgtcaggat ggccttctgc ttagtttgat gcctggcagt ttatggcggg cgtcctgccc
     3001 gccaccctcc gggccgttgc ttcacaacgt tcaaatccgc tcccggcgga tttgtcctac
     3061 tcaggagagc gttcaccgac aaacaacaga taaaacgaaa ggcccagtct tccgactgag
     3121 cctttcgttt tatttgatgc ctggcagttc cctactctcg cgttaacgct agcatggatg
     3181 ttttcccagt cacgacgttg taaaacgacg gccagtctta agctcgggcc ctgcagctct
     3241 agagctcgaa ttctacaggt cactaatacc atctaagtag ttggtt
//
