LOCUS       sequence-223880-        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"
     protein_bind    complement(1..124)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(652..747)
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(784..803)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(785..803)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(808..824)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(808..824)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(926..945)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             937..1746
                     /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(1004..1023)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      1893..2481
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2382..2401
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2635..2652
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2811..2838)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(2930..3016)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3065..3087
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3079..3096
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3080..3096
                     /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 gtggctggga agatgctttc caagatcttc ggcaacaagg agatgagaat attgatgctc
      181 ggacttgacg ccgccggaaa gaacacgatt ctgtacaaac tcaaactggg ccagtcggtt
      241 acgaccatcc cgacagtggg cttcaatgta gagactgtca cgtataaaaa cgtcaagttt
      301 aacgtttggg acgtcggcgg tcaggataag atccgtcctc tatggcggca ctactacacc
      361 ggcacgcagg gcctgatttt cgtagtggac tgtgcggatc gtgaccgcat cgatgaggct
      421 cggcaggagc tgcaccgcat tatcaacgat cgtgagatgc gcgacgccat tatcctgatc
      481 ttcgccaaca aacaagacct tccagacgcc atgaagccgc atgagataca ggagaagctg
      541 ggcctgacgc gcatccgaga tcgcaactgg tacgttcagc cgtcttgcgc tacaaccggc
      601 gacggactct acgaagggct cacctggctc acgtccaact acaaatccta acaactttat
      661 tatacaaagt tggcattata aaaaagcatt gcttatcaat ttgttgcaac gaacaggtca
      721 ctatcagtca aaataaaatc attatttgga gctccatggt agcgttaacg cggccgcgat
      781 atcccctata gtgagtcgta ttacatggtc atagctgttt cctggcagct ctggcccgtg
      841 tctcaaaatc tctgatgtta cattgcacaa gataaaaata tatcatcatg aacaataaaa
      901 ctgtctgctt acataaacag taatacaagg ggtgttatga gccatattca acgggaaacg
      961 tcgaggccgc gattaaattc caacatggat gctgatttat atgggtataa atgggctcgc
     1021 gataatgtcg ggcaatcagg tgcgacaatc tatcgcttgt atgggaagcc cgatgcgcca
     1081 gagttgtttc tgaaacatgg caaaggtagc gttgccaatg atgttacaga tgagatggtc
     1141 agactaaact ggctgacgga atttatgcct cttccgacca tcaagcattt tatccgtact
     1201 cctgatgatg catggttact caccactgcg atccccggaa aaacagcatt ccaggtatta
     1261 gaagaatatc ctgattcagg tgaaaatatt gttgatgcgc tggcagtgtt cctgcgccgg
     1321 ttgcattcga ttcctgtttg taattgtcct tttaacagcg atcgcgtatt tcgtctcgct
     1381 caggcgcaat cacgaatgaa taacggtttg gttgatgcga gtgattttga tgacgagcgt
     1441 aatggctggc ctgttgaaca agtctggaaa gaaatgcata aacttttgcc attctcaccg
     1501 gattcagtcg tcactcatgg tgatttctca cttgataacc ttatttttga cgaggggaaa
     1561 ttaataggtt gtattgatgt tggacgagtc ggaatcgcag accgatacca ggatcttgcc
     1621 atcctatgga actgcctcgg tgagttttct ccttcattac agaaacggct ttttcaaaaa
     1681 tatggtattg ataatcctga tatgaataaa ttgcagtttc atttgatgct cgatgagttt
     1741 ttctaatcag aattggttaa ttggttgtaa cactggcaga gcattacgct gacttgacgg
     1801 gacggcgcaa gctcatgacc aaaatccctt aacgtgagtt acgcgtcgtt ccactgagcg
     1861 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     1921 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     1981 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     2041 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     2101 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     2161 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     2221 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     2281 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     2341 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     2401 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     2461 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     2521 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     2581 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     2641 tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg
     2701 ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc
     2761 aacgcaatta atacgcgtac cgctagccag gaagagtttg tagaaacgca aaaaggccat
     2821 ccgtcaggat ggccttctgc ttagtttgat gcctggcagt ttatggcggg cgtcctgccc
     2881 gccaccctcc gggccgttgc ttcacaacgt tcaaatccgc tcccggcgga tttgtcctac
     2941 tcaggagagc gttcaccgac aaacaacaga taaaacgaaa ggcccagtct tccgactgag
     3001 cctttcgttt tatttgatgc ctggcagttc cctactctcg cgttaacgct agcatggatg
     3061 ttttcccagt cacgacgttg taaaacgacg gccagtctta agctcgggcc ctgcagctct
     3121 agagctcgaa ttctacaggt cactaatacc atctaagtag ttggtt
//
