LOCUS       sequence-223887-        3346 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Multisite gateway vector for 3' tagging with rab12.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3346)
  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 3346)
  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-rab12
FEATURES             Location/Qualifiers
     source          1..3346
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(732..827)
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(864..883)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(865..883)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(888..904)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(888..904)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1006..1025)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1017..1826
                     /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(1084..1103)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      1973..2561
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2462..2481
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2715..2732
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2891..2918)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3010..3096)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3145..3167
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3159..3176
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3160..3176
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(join(3247..3346,1..24))
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
ORIGIN
        1 gttcaacttt cttgtacaaa gtggctgagt cgggcttcgg ggtccaacgg agggccggtg
       61 gaggctctcc cgctgtggga gcttcgcagc gccgcaggaa gattccgccc agagccgctg
      121 attataaact gcaggtgatc attattggct ccagaggtgt ggggaaaacc agcctgatgg
      181 agagattcac ggatgacact ttttgtgagg cgtgcaagtc cactgtgggt gtggatttta
      241 aaatcaagac agtggagctg agaggaaaga agatcaggtt gcagatctgg gacacggcag
      301 gtcaggagcg gtttaacagc atcacttcag cgtattaccg cggtgccaaa ggcattgttt
      361 tggtttatga catcaccaag caggaaactt ttgaagacct gcctaaatgg atgaagatga
      421 tagacaagta tgcgtcagag gatgctgagc tcctattggt tgggaacaaa ctggactgcg
      481 agtctgatcg cgccatctca cggcaacaag ctgagaggtt tgcttcacgg atatcaggga
      541 tgcgtttctg tgaggccagt gccaaggata actttaacgt ggatgagatc ttccttaaac
      601 tagttgatga catcttgagt aagatgccac ttgaagtccc cagtaaggag ctttccaaca
      661 gtgttttgtc tcttcaaccc gagcctgaaa tcccaccaga gctgcctccg ccacgcatgc
      721 gctgctgttg acaactttat tatacaaagt tggcattata aaaaagcatt gcttatcaat
      781 ttgttgcaac gaacaggtca ctatcagtca aaataaaatc attatttgga gctccatggt
      841 agcgttaacg cggccgcgat atcccctata gtgagtcgta ttacatggtc atagctgttt
      901 cctggcagct ctggcccgtg tctcaaaatc tctgatgtta cattgcacaa gataaaaata
      961 tatcatcatg aacaataaaa ctgtctgctt acataaacag taatacaagg ggtgttatga
     1021 gccatattca acgggaaacg tcgaggccgc gattaaattc caacatggat gctgatttat
     1081 atgggtataa atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc tatcgcttgt
     1141 atgggaagcc cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc gttgccaatg
     1201 atgttacaga tgagatggtc agactaaact ggctgacgga atttatgcct cttccgacca
     1261 tcaagcattt tatccgtact cctgatgatg catggttact caccactgcg atccccggaa
     1321 aaacagcatt ccaggtatta gaagaatatc ctgattcagg tgaaaatatt gttgatgcgc
     1381 tggcagtgtt cctgcgccgg ttgcattcga ttcctgtttg taattgtcct tttaacagcg
     1441 atcgcgtatt tcgtctcgct caggcgcaat cacgaatgaa taacggtttg gttgatgcga
     1501 gtgattttga tgacgagcgt aatggctggc ctgttgaaca agtctggaaa gaaatgcata
     1561 aacttttgcc attctcaccg gattcagtcg tcactcatgg tgatttctca cttgataacc
     1621 ttatttttga cgaggggaaa ttaataggtt gtattgatgt tggacgagtc ggaatcgcag
     1681 accgatacca ggatcttgcc atcctatgga actgcctcgg tgagttttct ccttcattac
     1741 agaaacggct ttttcaaaaa tatggtattg ataatcctga tatgaataaa ttgcagtttc
     1801 atttgatgct cgatgagttt ttctaatcag aattggttaa ttggttgtaa cactggcaga
     1861 gcattacgct gacttgacgg gacggcgcaa gctcatgacc aaaatccctt aacgtgagtt
     1921 acgcgtcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     1981 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     2041 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     2101 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     2161 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     2221 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     2281 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     2341 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     2401 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     2461 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     2521 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     2581 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     2641 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     2701 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac
     2761 cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact
     2821 ggaaagcggg cagtgagcgc aacgcaatta atacgcgtac cgctagccag gaagagtttg
     2881 tagaaacgca aaaaggccat ccgtcaggat ggccttctgc ttagtttgat gcctggcagt
     2941 ttatggcggg cgtcctgccc gccaccctcc gggccgttgc ttcacaacgt tcaaatccgc
     3001 tcccggcgga tttgtcctac tcaggagagc gttcaccgac aaacaacaga taaaacgaaa
     3061 ggcccagtct tccgactgag cctttcgttt tatttgatgc ctggcagttc cctactctcg
     3121 cgttaacgct agcatggatg ttttcccagt cacgacgttg taaaacgacg gccagtctta
     3181 agctcgggcc ctgcagctct agagctcgaa ttctacaggt cactaatacc atctaagtag
     3241 ttggttcata gtgactgcat atgttgtgtt ttacagtatt atgtagtctg ttttttatgc
     3301 aaaatctaat ttaatatatt gatatttata tcattttacg tttctc
//
