LOCUS       sequence-224167-        3313 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Multisite gateway vector for 3' tagging with Rab27b.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3313)
  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 3313)
  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-Rab27b
FEATURES             Location/Qualifiers
     source          1..3313
                     /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(799..894)
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(931..950)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(932..950)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(955..971)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(955..971)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1073..1092)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1084..1893
                     /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(1151..1170)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2040..2628
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2529..2548
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2782..2799
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2958..2985)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3077..3163)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3212..3234
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3226..3243
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3227..3243
                     /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 gtggtgttca gctccagcag cactatgacc gatggagatt acgattatct gatcaaactc
      181 ctggctttgg gggactccgg ggtcggcaag accactttcc tctaccgata cacagataac
      241 aagttcaacc ccaaattcat caccaccgtg ggcatcgact tcagggaaaa gagagtggtt
      301 tatacgacta acagccccaa tgggacgacg gggaaaacct ttaaggttca cctgcagctg
      361 tgggacactg cgggacagga gaggttcagg agtttgacca ctgcattctt cagggatgcc
      421 atgggttttc tgctgatgtt tgatctgacc agccagcaga gctttctcaa cgtcaggaac
      481 tggatgagtc agctccaggc taatgcatat tgtgagaatc cagatatcgt gctggtggga
      541 aataaagcag atctagccga ccagagagaa gttcaggaga aacaagcgaa agaactcgca
      601 gacaaatacg gcatcccgta cttcgagacg agtgcagcga cgggctctga ggtggacaag
      661 gctgtggtga cgctgctgga tctggtgatg aagcggatgg agcagtgtgt ggagaagccc
      721 agcgcagacg caaacaccag cgacggcagc agcaaactga gcgccgcacc tgcccaacag
      781 aagaagtgtg catgctgaca actttattat acaaagttgg cattataaaa aagcattgct
      841 tatcaatttg ttgcaacgaa caggtcacta tcagtcaaaa taaaatcatt atttggagct
      901 ccatggtagc gttaacgcgg ccgcgatatc ccctatagtg agtcgtatta catggtcata
      961 gctgtttcct ggcagctctg gcccgtgtct caaaatctct gatgttacat tgcacaagat
     1021 aaaaatatat catcatgaac aataaaactg tctgcttaca taaacagtaa tacaaggggt
     1081 gttatgagcc atattcaacg ggaaacgtcg aggccgcgat taaattccaa catggatgct
     1141 gatttatatg ggtataaatg ggctcgcgat aatgtcgggc aatcaggtgc gacaatctat
     1201 cgcttgtatg ggaagcccga tgcgccagag ttgtttctga aacatggcaa aggtagcgtt
     1261 gccaatgatg ttacagatga gatggtcaga ctaaactggc tgacggaatt tatgcctctt
     1321 ccgaccatca agcattttat ccgtactcct gatgatgcat ggttactcac cactgcgatc
     1381 cccggaaaaa cagcattcca ggtattagaa gaatatcctg attcaggtga aaatattgtt
     1441 gatgcgctgg cagtgttcct gcgccggttg cattcgattc ctgtttgtaa ttgtcctttt
     1501 aacagcgatc gcgtatttcg tctcgctcag gcgcaatcac gaatgaataa cggtttggtt
     1561 gatgcgagtg attttgatga cgagcgtaat ggctggcctg ttgaacaagt ctggaaagaa
     1621 atgcataaac ttttgccatt ctcaccggat tcagtcgtca ctcatggtga tttctcactt
     1681 gataacctta tttttgacga ggggaaatta ataggttgta ttgatgttgg acgagtcgga
     1741 atcgcagacc gataccagga tcttgccatc ctatggaact gcctcggtga gttttctcct
     1801 tcattacaga aacggctttt tcaaaaatat ggtattgata atcctgatat gaataaattg
     1861 cagtttcatt tgatgctcga tgagtttttc taatcagaat tggttaattg gttgtaacac
     1921 tggcagagca ttacgctgac ttgacgggac ggcgcaagct catgaccaaa atcccttaac
     1981 gtgagttacg cgtcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     2041 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     2101 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     2161 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     2221 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     2281 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     2341 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     2401 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     2461 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     2521 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     2581 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     2641 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     2701 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     2761 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata
     2821 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt
     2881 cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc tagccaggaa
     2941 gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta gtttgatgcc
     3001 tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc acaacgttca
     3061 aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa caacagataa
     3121 aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg gcagttccct
     3181 actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa aacgacggcc
     3241 agtcttaagc tcgggccctg cagctctaga gctcgaattc tacaggtcac taataccatc
     3301 taagtagttg gtt
//
