LOCUS       sequence-223973-        3254 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Multisite gateway vector for 3' tagging with rab35b.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3254)
  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 3254)
  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-rab35b
FEATURES             Location/Qualifiers
     source          1..3254
                     /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(640..735)
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(772..791)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(773..791)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(796..812)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(796..812)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(914..933)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             925..1734
                     /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(992..1011)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      1881..2469
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2370..2389
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2623..2640
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2799..2826)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(2918..3004)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3053..3075
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3067..3084
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3068..3084
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gttcaacttt cttgtacaaa gtggtggccc gcgactacga tcacctcttc aagctgctga
       61 tcatcggtga cagcggtgtt ggaaagagca gtcttctcct gcgtttcgcc gacaacacat
      121 tttcaggcag ttacatcacc accataggag tcgacttcaa gatccgcact gtggagctta
      181 atggagagaa ggtgaaactg cagatatggg acacagccgg acaagagcga ttccgaacga
      241 ttacatctac atattacaga ggaacccatg gtgtgatagt agtgtatgac gttaccagcg
      301 ccgagtcctt tgtcaatgtt aaacgatggc tacatgaaat aaaccagaac tgtgatgacg
      361 tctgtcgaat attagtgggc aataaaaatg atgatccaaa ctctaaggtg gtggagacga
      421 atgatgcaca gaagtttgca gaacagatgg gcatcagtct gttcgagaca agtgcgaaag
      481 aaaatgtcaa tgtggaggag atgtttaact gcatcacaga gttggttctc cgggcaaaga
      541 aagagtctgt ggccaaacag cagcagcagc agcagaacga tgtgattaaa ctcaagcaga
      601 acagcaaacg gaagaagaag tgctgctagt ggacgaacac aactttatta tacaaagttg
      661 gcattataaa aaagcattgc ttatcaattt gttgcaacga acaggtcact atcagtcaaa
      721 ataaaatcat tatttggagc tccatggtag cgttaacgcg gccgcgatat cccctatagt
      781 gagtcgtatt acatggtcat agctgtttcc tggcagctct ggcccgtgtc tcaaaatctc
      841 tgatgttaca ttgcacaaga taaaaatata tcatcatgaa caataaaact gtctgcttac
      901 ataaacagta atacaagggg tgttatgagc catattcaac gggaaacgtc gaggccgcga
      961 ttaaattcca acatggatgc tgatttatat gggtataaat gggctcgcga taatgtcggg
     1021 caatcaggtg cgacaatcta tcgcttgtat gggaagcccg atgcgccaga gttgtttctg
     1081 aaacatggca aaggtagcgt tgccaatgat gttacagatg agatggtcag actaaactgg
     1141 ctgacggaat ttatgcctct tccgaccatc aagcatttta tccgtactcc tgatgatgca
     1201 tggttactca ccactgcgat ccccggaaaa acagcattcc aggtattaga agaatatcct
     1261 gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt gcattcgatt
     1321 cctgtttgta attgtccttt taacagcgat cgcgtatttc gtctcgctca ggcgcaatca
     1381 cgaatgaata acggtttggt tgatgcgagt gattttgatg acgagcgtaa tggctggcct
     1441 gttgaacaag tctggaaaga aatgcataaa cttttgccat tctcaccgga ttcagtcgtc
     1501 actcatggtg atttctcact tgataacctt atttttgacg aggggaaatt aataggttgt
     1561 attgatgttg gacgagtcgg aatcgcagac cgataccagg atcttgccat cctatggaac
     1621 tgcctcggtg agttttctcc ttcattacag aaacggcttt ttcaaaaata tggtattgat
     1681 aatcctgata tgaataaatt gcagtttcat ttgatgctcg atgagttttt ctaatcagaa
     1741 ttggttaatt ggttgtaaca ctggcagagc attacgctga cttgacggga cggcgcaagc
     1801 tcatgaccaa aatcccttaa cgtgagttac gcgtcgttcc actgagcgtc agaccccgta
     1861 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     1921 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     1981 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     2041 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     2101 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     2161 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     2221 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     2281 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     2341 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     2401 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     2461 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     2521 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     2581 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     2641 gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa
     2701 tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat
     2761 acgcgtaccg ctagccagga agagtttgta gaaacgcaaa aaggccatcc gtcaggatgg
     2821 ccttctgctt agtttgatgc ctggcagttt atggcgggcg tcctgcccgc caccctccgg
     2881 gccgttgctt cacaacgttc aaatccgctc ccggcggatt tgtcctactc aggagagcgt
     2941 tcaccgacaa acaacagata aaacgaaagg cccagtcttc cgactgagcc tttcgtttta
     3001 tttgatgcct ggcagttccc tactctcgcg ttaacgctag catggatgtt ttcccagtca
     3061 cgacgttgta aaacgacggc cagtcttaag ctcgggccct gcagctctag agctcgaatt
     3121 ctacaggtca ctaataccat ctaagtagtt ggttcatagt gactgcatat gttgtgtttt
     3181 acagtattat gtagtctgtt ttttatgcaa aatctaattt aatatattga tatttatatc
     3241 attttacgtt tctc
//
