LOCUS       sequence-206509-        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"
     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(3015..3110)
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3147..3166)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3148..3166)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3171..3187)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3171..3187)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3289..3308)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3300..4109
                     /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(3367..3386)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4256..4844
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4745..4764
                     /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 gcaaccgggg gatggaggaa ttgatccccc
      781 tgatcaataa actccaagat gctttcagct ccattggcca aagttgtaac ctggatctgc
      841 cgcagatagc agttgtcgga ggacagagcg ccggagctag ctccgtgctg gaaaatttcg
      901 tcggcaggga tttccttccg cgaggctcag gcatcgtcac ccgcaggcct ctcattctgc
      961 agctagtaaa caacaaagcc gaatatgcag agttcctaca ctgcaaaggg aggaagttcg
     1021 tggactttga tgaagtccgg caggagattg aagcagaaac cgacaggatt acaggctcca
     1081 ataagggcat ctctcctatc cccatcaacc tgagagtcta ctctcctaac gtattgaatc
     1141 tgacactcat cgatttgcct gggatgacga aggtggcagt gggcgaccag ccgcctgaca
     1201 tcgagcacca gattcgggac atgataatgc agtttatcac cagggagagc tgtctaatcc
     1261 ttgccgtcac cccagccaac atggacttgg ccaattcgga tgccctcaag gtggcgaaag
     1321 aagtggaccc tcaaggactg cgtaccattg gtgtaatcac taaactggat ttaatggatg
     1381 agggtacaga tgcccgggat atcctggaaa acaaactttt accacttcgt agaggttata
     1441 tcggtgtagt gaaccggagt cagaaagaca ttgatggccg taaggacatt cgtgctgcgc
     1501 tggcagcaga aaggaagttt ttcctgtccc accctagtta cagacatatg gctgaaagga
     1561 tgggaacgcc acacctgcaa aaagccctca accagcaatt gaccaaccac atccgggaca
     1621 cacttcccgg cttgcggagc aaactccagt ctcagcttct ctctctagaa aaagaagtgg
     1681 aagagtataa gaacttccgc ccagatgacc ctactcgaaa gaccaaggcc ctgctgcaga
     1741 tggtccagca atttggtgtg gactttgaga agtgcattga gggatccgga gatcaggtgg
     1801 acactgcaga gctgtctgga ggtgccagga ttaaccgaat ctttcatgag cgcttcccct
     1861 ttgagctggt caagatcgtg tttgatgaaa aggagctccg gagagaaatt agtcacgcca
     1921 ttaagaacgt ccatggtgtc agaactgggc tgttcactcc agatctggcg tttgaagcca
     1981 tcgtgaaaaa gcagatcatt aagctgaaag agccctgtct gaaatgtatc gatctggtta
     2041 ttcaggagct catcaacaca ttcaggcagt gcaccaacaa gttaagctcg taccctagac
     2101 tccgagaaga gacggagagg atcgtcacta cttacgtcag agagagagaa gggaagacca
     2161 aggaccaggt tcttttgctg attgacactg agctgtcata catcaacacc aaccacgagg
     2221 acttcattgg atttgcaaat gcacagcaaa ggaatatcgt caacaagaag agggccattc
     2281 ccaaccaggt gatccgtaaa ggctggttaa ccctgaacat cagcatcatg aagggaggtt
     2341 caaaggagta ctggttcgtc ctgactgccg aatcgctgtc ctggtacaaa gacgaggagg
     2401 aaaaagagaa gaaatacatg cttcctctgg ataacctgaa gctcagagac gtggagaaag
     2461 gcttcatgtc cactaagcac atttttgcaa tcttcaacac tgagcaaagg aatgtgtata
     2521 aggacttgcg tcagatcgag ctggcctgtg actcgcagga ggacatggat agttggaaag
     2581 cctcttttct cagagcaggc gtttaccctg agaaagatca agtggagtca gaagaggcag
     2641 ccccagccga cagcttctcc atggaccctc agttggaaag acaggtcgag accatccgta
     2701 acctggtgga ctcctatatt ggaatcgtca acaaaaccat cagagacctc atgcccaaga
     2761 ctatcatgca tctcatgatc aacagcgcta aagacttcat ccactcagag ctgctggcgt
     2821 acctgtactc gtctggagat cagaacagtc tgatggagga gtctgcagac caggcccagc
     2881 gcagagacga gatgctgagg atgtaccatg ccatcaaaga ggcactcagc atcatcggtg
     2941 acatcagcac cagcaccatt tccacacctg tgccaccacc cgtcaacgac agctggatac
     3001 ctgaaaccag gtgacaactt tattatacaa agttggcatt ataaaaaagc attgcttatc
     3061 aatttgttgc aacgaacagg tcactatcag tcaaaataaa atcattattt ggagctccat
     3121 ggtagcgtta acgcggccgc gatatcccct atagtgagtc gtattacatg gtcatagctg
     3181 tttcctggca gctctggccc gtgtctcaaa atctctgatg ttacattgca caagataaaa
     3241 atatatcatc atgaacaata aaactgtctg cttacataaa cagtaataca aggggtgtta
     3301 tgagccatat tcaacgggaa acgtcgaggc cgcgattaaa ttccaacatg gatgctgatt
     3361 tatatgggta taaatgggct cgcgataatg tcgggcaatc aggtgcgaca atctatcgct
     3421 tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca
     3481 atgatgttac agatgagatg gtcagactaa actggctgac ggaatttatg cctcttccga
     3541 ccatcaagca ttttatccgt actcctgatg atgcatggtt actcaccact gcgatccccg
     3601 gaaaaacagc attccaggta ttagaagaat atcctgattc aggtgaaaat attgttgatg
     3661 cgctggcagt gttcctgcgc cggttgcatt cgattcctgt ttgtaattgt ccttttaaca
     3721 gcgatcgcgt atttcgtctc gctcaggcgc aatcacgaat gaataacggt ttggttgatg
     3781 cgagtgattt tgatgacgag cgtaatggct ggcctgttga acaagtctgg aaagaaatgc
     3841 ataaactttt gccattctca ccggattcag tcgtcactca tggtgatttc tcacttgata
     3901 accttatttt tgacgagggg aaattaatag gttgtattga tgttggacga gtcggaatcg
     3961 cagaccgata ccaggatctt gccatcctat ggaactgcct cggtgagttt tctccttcat
     4021 tacagaaacg gctttttcaa aaatatggta ttgataatcc tgatatgaat aaattgcagt
     4081 ttcatttgat gctcgatgag tttttctaat cagaattggt taattggttg taacactggc
     4141 agagcattac gctgacttga cgggacggcg caagctcatg accaaaatcc cttaacgtga
     4201 gttacgcgtc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     4261 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     4321 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     4381 gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga
     4441 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     4501 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     4561 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     4621 ccgaactgag atacctacag cgtgagcatt gagaaagcgc cacgcttccc gaagggagaa
     4681 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     4741 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     4801 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     4861 cctttttacg gttcctggcc ttttgctggc cttttgctca catgtt
//
