LOCUS       sequence-222102-        4906 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Multisite gateway vector for 3' tagging with dominant negative 
            Dynamin2a.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4906)
  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 4906)
  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-Dnm2a-DN
FEATURES             Location/Qualifiers
     source          1..4906
                     /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(2392..2487)
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2524..2543)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2525..2543)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2548..2564)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2548..2564)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2666..2685)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2677..3486
                     /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(2744..2763)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3633..4221
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4122..4141
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4375..4392
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(4551..4578)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4670..4756)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     4805..4827
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4819..4836
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4820..4836
                     /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 gtggctggca accgggggat ggaggaattg atccccctga tcaataaact ccaagatgct
      181 ttcagctcca ttggccaaag ttgtaacctg gatctgccgc agatagcagt tgtcggagga
      241 cagagcgccg gagctagctc cgtgctggaa aatttcgtcg gcagggattt ccttccgcga
      301 ggctcaggca tcgtcacccg caggcctctc attctgcagc tagtaaacaa caaagccgaa
      361 tatgcagagt tcctacactg caaagggagg aagttcgtgg actttgatga agtccggcag
      421 gagattgaag cagaaaccga caggattaca ggctccaata agggcatctc tcctatcccc
      481 atcaacctga gagtctactc tcctaacgta ttgaatctga cactcatcga tttgcctggg
      541 atgacgaagg tggcagtggg cgaccagccg cctgacatcg agcaccagat tcgggacatg
      601 ataatgcagt ttatcaccag ggagagctgt ctaatccttg ccgtcacccc agccaacatg
      661 gacttggcca attcggatgc cctcaaggtg gcgaaagaag tggaccctca aggactgcgt
      721 accattggtg taatcactaa actggattta atggatgagg gtacagatgc ccgggatatc
      781 ctggaaaaca aacttttacc acttcgtaga ggttatatcg gtgtagtgaa ccggagtcag
      841 aaagacattg atggccgtaa ggacattcgt gctgcgctgg cagcagaaag gaagtttttc
      901 ctgtcccacc ctagttacag acatatggct gaaaggatgg gaacgccaca cctgcaaaaa
      961 gccctcaacc agcaattgac caaccacatc cgggacacac ttcccggctt gcggagcaaa
     1021 ctccagtctc agcttctctc tctagaaaaa gaagtggaag agtataagaa cttccgccca
     1081 gatgacccta ctcgaaagac caaggccctg ctgcagatgg tccagcaatt tggtgtggac
     1141 tttgagaagt gcattgaggg atccggagat caggtggaca ctgcagagct gtctggaggt
     1201 gccaggatta accgaatctt tcatgagcgc ttcccctttg agctggtcaa gatcgtgttt
     1261 gatgaaaagg agctccggag agaaattagt cacgccatta agaacgtcca tggtgtcaga
     1321 actgggctgt tcactccaga tctggcgttt gaagccatcg tgaaaaagca gatcattaag
     1381 ctgaaagagc cctgtctgaa atgtatcgat ctggttattc aggagctcat caacacattc
     1441 aggcagtgca ccaacaagtt aagctcgtac cctagactcc gagaagagac ggagaggatc
     1501 gtcactactt acgtcagaga gagagaaggg aagaccaagg accaggttct tttgctgatt
     1561 gacactgagc tgtcatacat caacaccaac cacgaggact tcattggatt tgcaaatgca
     1621 cagcaaagga atatcgtcaa caagaagagg gccattccca accaggtgat ccgtaaaggc
     1681 tggttaaccc tgaacatcag catcatgaag ggaggttcaa aggagtactg gttcgtcctg
     1741 actgccgaat cgctgtcctg gtacaaagac gaggaggaaa aagagaagaa atacatgctt
     1801 cctctggata acctgaagct cagagacgtg gagaaaggct tcatgtccac taagcacatt
     1861 tttgcaatct tcaacactga gcaaaggaat gtgtataagg acttgcgtca gatcgagctg
     1921 gcctgtgact cgcaggagga catggatagt tggaaagcct cttttctcag agcaggcgtt
     1981 taccctgaga aagatcaagt ggagtcagaa gaggcagccc cagccgacag cttctccatg
     2041 gaccctcagt tggaaagaca ggtcgagacc atccgtaacc tggtggactc ctatattgga
     2101 atcgtcaaca aaaccatcag agacctcatg cccaagacta tcatgcatct catgatcaac
     2161 agcgctaaag acttcatcca ctcagagctg ctggcgtacc tgtactcgtc tggagatcag
     2221 aacagtctga tggaggagtc tgcagaccag gcccagcgca gagacgagat gctgaggatg
     2281 taccatgcca tcaaagaggc actcagcatc atcggtgaca tcagcaccag caccatttcc
     2341 acacctgtgc caccacccgt caacgacagc tggatacctg aaaccaggtg acaactttat
     2401 tatacaaagt tggcattata aaaaagcatt gcttatcaat ttgttgcaac gaacaggtca
     2461 ctatcagtca aaataaaatc attatttgga gctccatggt agcgttaacg cggccgcgat
     2521 atcccctata gtgagtcgta ttacatggtc atagctgttt cctggcagct ctggcccgtg
     2581 tctcaaaatc tctgatgtta cattgcacaa gataaaaata tatcatcatg aacaataaaa
     2641 ctgtctgctt acataaacag taatacaagg ggtgttatga gccatattca acgggaaacg
     2701 tcgaggccgc gattaaattc caacatggat gctgatttat atgggtataa atgggctcgc
     2761 gataatgtcg ggcaatcagg tgcgacaatc tatcgcttgt atgggaagcc cgatgcgcca
     2821 gagttgtttc tgaaacatgg caaaggtagc gttgccaatg atgttacaga tgagatggtc
     2881 agactaaact ggctgacgga atttatgcct cttccgacca tcaagcattt tatccgtact
     2941 cctgatgatg catggttact caccactgcg atccccggaa aaacagcatt ccaggtatta
     3001 gaagaatatc ctgattcagg tgaaaatatt gttgatgcgc tggcagtgtt cctgcgccgg
     3061 ttgcattcga ttcctgtttg taattgtcct tttaacagcg atcgcgtatt tcgtctcgct
     3121 caggcgcaat cacgaatgaa taacggtttg gttgatgcga gtgattttga tgacgagcgt
     3181 aatggctggc ctgttgaaca agtctggaaa gaaatgcata aacttttgcc attctcaccg
     3241 gattcagtcg tcactcatgg tgatttctca cttgataacc ttatttttga cgaggggaaa
     3301 ttaataggtt gtattgatgt tggacgagtc ggaatcgcag accgatacca ggatcttgcc
     3361 atcctatgga actgcctcgg tgagttttct ccttcattac agaaacggct ttttcaaaaa
     3421 tatggtattg ataatcctga tatgaataaa ttgcagtttc atttgatgct cgatgagttt
     3481 ttctaatcag aattggttaa ttggttgtaa cactggcaga gcattacgct gacttgacgg
     3541 gacggcgcaa gctcatgacc aaaatccctt aacgtgagtt acgcgtcgtt ccactgagcg
     3601 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     3661 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     3721 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     3781 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     3841 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     3901 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     3961 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     4021 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     4081 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     4141 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     4201 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     4261 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     4321 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     4381 tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg
     4441 ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc
     4501 aacgcaatta atacgcgtac cgctagccag gaagagtttg tagaaacgca aaaaggccat
     4561 ccgtcaggat ggccttctgc ttagtttgat gcctggcagt ttatggcggg cgtcctgccc
     4621 gccaccctcc gggccgttgc ttcacaacgt tcaaatccgc tcccggcgga tttgtcctac
     4681 tcaggagagc gttcaccgac aaacaacaga taaaacgaaa ggcccagtct tccgactgag
     4741 cctttcgttt tatttgatgc ctggcagttc cctactctcg cgttaacgct agcatggatg
     4801 ttttcccagt cacgacgttg taaaacgacg gccagtctta agctcgggcc ctgcagctct
     4861 agagctcgaa ttctacaggt cactaatacc atctaagtag ttggtt
//
