LOCUS       sequence-223967-        3274 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Multisite gateway vector for 3' tagging with rab32b.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3274)
  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 3274)
  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-rab32b
FEATURES             Location/Qualifiers
     source          1..3274
                     /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(760..855)
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(892..911)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(893..911)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(916..932)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(916..932)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1034..1053)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1045..1854
                     /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(1112..1131)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2001..2589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2490..2509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2743..2760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2919..2946)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3038..3124)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3173..3195
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3187..3204
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3188..3204
                     /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 gtggtgagga aaatgtcagg agactcagag ttaggatgca ctgaacattt atttaaagtt
      181 ttggtgattg gagaccataa agttgggaag agcagctttg ttaaactcta tgtgaggaat
      241 cagttttacg aggagttgaa aaccagtata ggtgtggatt tttctatgaa ggttatcgat
      301 tgggacgctc acacacgagt gagattgcag ttttgggata tagctggtca agagcgtgtg
      361 cgaggactga atcgtgtgta cttcagggga cccagtggtg cttttgtagt atacgacctg
      421 acaagcggct catcgctgga tggggctctg aactggaaac gtgaactcga ttgtcaagta
      481 atgttgaaaa atggccatcc gatccctgct gttctgctgg ctaataaatg tgatgagact
      541 aaagtgagcc ggaggaacgc atcccttctg gagaagctct gccaggagaa gggttttact
      601 gggtgctttc agacttcagc aaaggagaat atcaatgtgg atgaagctgc taaatttctt
      661 gtgaagcaca ttctccaaat taacgaagaa tcaatgaaag aagaagagca gaagattcac
      721 ctcaaaccac acacaaacac aaactcggtc tgctgctgac aactttatta tacaaagttg
      781 gcattataaa aaagcattgc ttatcaattt gttgcaacga acaggtcact atcagtcaaa
      841 ataaaatcat tatttggagc tccatggtag cgttaacgcg gccgcgatat cccctatagt
      901 gagtcgtatt acatggtcat agctgtttcc tggcagctct ggcccgtgtc tcaaaatctc
      961 tgatgttaca ttgcacaaga taaaaatata tcatcatgaa caataaaact gtctgcttac
     1021 ataaacagta atacaagggg tgttatgagc catattcaac gggaaacgtc gaggccgcga
     1081 ttaaattcca acatggatgc tgatttatat gggtataaat gggctcgcga taatgtcggg
     1141 caatcaggtg cgacaatcta tcgcttgtat gggaagcccg atgcgccaga gttgtttctg
     1201 aaacatggca aaggtagcgt tgccaatgat gttacagatg agatggtcag actaaactgg
     1261 ctgacggaat ttatgcctct tccgaccatc aagcatttta tccgtactcc tgatgatgca
     1321 tggttactca ccactgcgat ccccggaaaa acagcattcc aggtattaga agaatatcct
     1381 gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt gcattcgatt
     1441 cctgtttgta attgtccttt taacagcgat cgcgtatttc gtctcgctca ggcgcaatca
     1501 cgaatgaata acggtttggt tgatgcgagt gattttgatg acgagcgtaa tggctggcct
     1561 gttgaacaag tctggaaaga aatgcataaa cttttgccat tctcaccgga ttcagtcgtc
     1621 actcatggtg atttctcact tgataacctt atttttgacg aggggaaatt aataggttgt
     1681 attgatgttg gacgagtcgg aatcgcagac cgataccagg atcttgccat cctatggaac
     1741 tgcctcggtg agttttctcc ttcattacag aaacggcttt ttcaaaaata tggtattgat
     1801 aatcctgata tgaataaatt gcagtttcat ttgatgctcg atgagttttt ctaatcagaa
     1861 ttggttaatt ggttgtaaca ctggcagagc attacgctga cttgacggga cggcgcaagc
     1921 tcatgaccaa aatcccttaa cgtgagttac gcgtcgttcc actgagcgtc agaccccgta
     1981 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     2041 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     2101 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     2161 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     2221 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     2281 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     2341 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     2401 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     2461 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     2521 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     2581 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     2641 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     2701 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     2761 gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa
     2821 tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat
     2881 acgcgtaccg ctagccagga agagtttgta gaaacgcaaa aaggccatcc gtcaggatgg
     2941 ccttctgctt agtttgatgc ctggcagttt atggcgggcg tcctgcccgc caccctccgg
     3001 gccgttgctt cacaacgttc aaatccgctc ccggcggatt tgtcctactc aggagagcgt
     3061 tcaccgacaa acaacagata aaacgaaagg cccagtcttc cgactgagcc tttcgtttta
     3121 tttgatgcct ggcagttccc tactctcgcg ttaacgctag catggatgtt ttcccagtca
     3181 cgacgttgta aaacgacggc cagtcttaag ctcgggccct gcagctctag agctcgaatt
     3241 ctacaggtca ctaataccat ctaagtagtt ggtt
//
