LOCUS       sequence-206529-        3166 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Multisite gateway vector for 3' tagging with dominant negative ARF6.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3166)
  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 3166)
  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-ARF6-DN
FEATURES             Location/Qualifiers
     source          1..3166
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     536..553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(624..747)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(1275..1370)
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1407..1426)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1408..1426)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1431..1447)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1431..1447)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1549..1568)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1560..2369
                     /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(1627..1646)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2516..3104
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3005..3024
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggccctg cagctctaga gctcgaattc tacaggtcac
      601 taataccatc taagtagttg gttcatagtg actgcatatg ttgtgtttta cagtattatg
      661 tagtctgttt tttatgcaaa atctaattta atatattgat atttatatca ttttacgttt
      721 ctcgttcaac tttcttgtac aaagtggctg ggaagatgct ttccaagatc ttcggcaaca
      781 aggagatgag aatattgatg ctcggacttg acgccgccgg aaagaacacg attctgtaca
      841 aactcaaact gggccagtcg gttacgacca tcccgacagt gggcttcaat gtagagactg
      901 tcacgtataa aaacgtcaag tttaacgttt gggacgtcgg cggtcaggat aagatccgtc
      961 ctctatggcg gcactactac accggcacgc agggcctgat tttcgtagtg gactgtgcgg
     1021 atcgtgaccg catcgatgag gctcggcagg agctgcaccg cattatcaac gatcgtgaga
     1081 tgcgcgacgc cattatcctg atcttcgcca acaaacaaga ccttccagac gccatgaagc
     1141 cgcatgagat acaggagaag ctgggcctga cgcgcatccg agatcgcaac tggtacgttc
     1201 agccgtcttg cgctacaacc ggcgacggac tctacgaagg gctcacctgg ctcacgtcca
     1261 actacaaatc ctaacaactt tattatacaa agttggcatt ataaaaaagc attgcttatc
     1321 aatttgttgc aacgaacagg tcactatcag tcaaaataaa atcattattt ggagctccat
     1381 ggtagcgtta acgcggccgc gatatcccct atagtgagtc gtattacatg gtcatagctg
     1441 tttcctggca gctctggccc gtgtctcaaa atctctgatg ttacattgca caagataaaa
     1501 atatatcatc atgaacaata aaactgtctg cttacataaa cagtaataca aggggtgtta
     1561 tgagccatat tcaacgggaa acgtcgaggc cgcgattaaa ttccaacatg gatgctgatt
     1621 tatatgggta taaatgggct cgcgataatg tcgggcaatc aggtgcgaca atctatcgct
     1681 tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca
     1741 atgatgttac agatgagatg gtcagactaa actggctgac ggaatttatg cctcttccga
     1801 ccatcaagca ttttatccgt actcctgatg atgcatggtt actcaccact gcgatccccg
     1861 gaaaaacagc attccaggta ttagaagaat atcctgattc aggtgaaaat attgttgatg
     1921 cgctggcagt gttcctgcgc cggttgcatt cgattcctgt ttgtaattgt ccttttaaca
     1981 gcgatcgcgt atttcgtctc gctcaggcgc aatcacgaat gaataacggt ttggttgatg
     2041 cgagtgattt tgatgacgag cgtaatggct ggcctgttga acaagtctgg aaagaaatgc
     2101 ataaactttt gccattctca ccggattcag tcgtcactca tggtgatttc tcacttgata
     2161 accttatttt tgacgagggg aaattaatag gttgtattga tgttggacga gtcggaatcg
     2221 cagaccgata ccaggatctt gccatcctat ggaactgcct cggtgagttt tctccttcat
     2281 tacagaaacg gctttttcaa aaatatggta ttgataatcc tgatatgaat aaattgcagt
     2341 ttcatttgat gctcgatgag tttttctaat cagaattggt taattggttg taacactggc
     2401 agagcattac gctgacttga cgggacggcg caagctcatg accaaaatcc cttaacgtga
     2461 gttacgcgtc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     2521 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     2581 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     2641 gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga
     2701 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     2761 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     2821 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     2881 ccgaactgag atacctacag cgtgagcatt gagaaagcgc cacgcttccc gaagggagaa
     2941 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     3001 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     3061 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     3121 cctttttacg gttcctggcc ttttgctggc cttttgctca catgtt
//
