LOCUS       sequence-206511-        5200 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Multisite gateway vector for 3' tagging with dominant negative 
            Dynamin2b.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5200)
  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 5200)
  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-Dnm2b-DN
FEATURES             Location/Qualifiers
     source          1..5200
                     /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..748)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(3309..3404)
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3441..3460)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3442..3460)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3465..3481)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3465..3481)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3583..3602)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3594..4403
                     /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(3661..3680)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4550..5138
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5039..5058
                     /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 attcatagtg actgcatatg ttgtgtttta cagtattatg
      661 tagtctgttt tttatgcaaa atctaattta atatattgat atttatatca ttttacgttt
      721 ctcgttcaac tttcttgtac aaagtggtgg ggaaccgcgg gatggaggac ttgattcccc
      781 tcattaacaa gctgcatgac gccttcagct ccatcggcca gagctgcaac ctggagctgc
      841 cgcagatcgc ggtggtcggc gtgcagagtg ccggcgcgac ctccgtgctc gagaacttcg
      901 tgggaaggga ctttctgccg agaggaccag gaatcgtcac tcgcaggcct ctgattctcc
      961 agctggtgaa cagtaaagct gaatatgccg agttcctgca ctgtaagggc cgcaagtttg
     1021 tggactttga cgaggttcgg atggagattg aagcagaaac agaccgaatc actgaatcta
     1081 acattggaat ctctgcagtg cccattaacc tgcgccttta ctcgcccaac gtgctgaatc
     1141 tgacgctgat cgatcttccg ggcataacga aggtggcggt gggtgaccag ccgcaggata
     1201 tcgagtacca gatccgggat atggtgttgc agttcatctc caaagagaac tgtctgattc
     1261 tggccatcac tccagcaaac atggacctga ccaactctga cgctctcaag atcagcaaag
     1321 aggtggaccc tcatggtctg cgcaccatcg gtgtgatcac caaactggac ctgatggatg
     1381 aaggaaccga cgccagagac atcctggaga acaagctgct gccgctgcgc agaggttata
     1441 tcggtgtggt gaaccgcagt cagaaggaca ttgatggcag gaaggacatc cgcgcggctc
     1501 tttctgctga gaggaagttc ttcctgtccc atccagcata cagacacata gcagagagaa
     1561 tgggcacacc gtacctgcag aaaacactca accagcaact gatgaaccac atccgcgaca
     1621 cactgccggc gctgcgcagt aaactgcagt ctcagctgct gtcgctggag aaagaggtgg
     1681 aggagtacaa gaacttcaaa ccagacgatc ctgcgcgcaa aaccaaagcc ctgctgcaga
     1741 tggtccagca gttcggtgtg gactttgaga agcgtatagg gggctctgga gatcaggtgg
     1801 acactctgga actctccggc ggagctcgaa taaaccgcat cttccacgag cgcttcccct
     1861 tcgagctggt gaagacggag tttgatgaga aggagttgcg gagggagatc agttacgcca
     1921 tcaagaacat tcacggagtc aggacggggc tcttcacccc agatctggcc ttcgaggcca
     1981 tcgtgaaaaa gcagctgata aagctgaagg agccgtgcct caagtgcatc gacctggtca
     2041 tccaggagct gatcaacacg gtcaggcaga gcacactgaa gctgaagttg tatcctcggc
     2101 tgagagaaga gacggagaga atcatcacaa tatatgtcag agagagagag accaaaacca
     2161 aagaccagat gatgctgctg atcgatgtcc agctctccta catcaacaca aaccacgagg
     2221 acttcatcgg attcgccaat gcgcaacaga gcagcagaga tgtcactaag aagcaagccg
     2281 ttcccaatca ggtgatccgc cgtggttggc tgaccatcaa catcagcatc atgaagggcg
     2341 gctccaaaga ctactggttc gtgctgacgg cggagtctct gtcctggtac aaggatgagg
     2401 aggagaagga gaagaagttc atgctgccgc tggataacct gaagctgcgg gatgtggaga
     2461 agagcttcat gagcaacaaa cacaccttcg gcatcttcaa caccgagcag aggaacgtgt
     2521 ataaagacct gcgtcagatc gagctggcct gcgccactca ggaggatgtg gacagctgga
     2581 aggcctcttt cctcagagcg ggagtctatc ctgagaaaga ccaggtggag aacggggacg
     2641 tgtcagcaga ctttctctcc atggaccctc agctggagcg tcaggttgag accatcagta
     2701 atctggtgga ctcgtacatc agcatcgtca acaagagcat ccgagacctg atgcccaaaa
     2761 ccatcatgca cctgatgatc aacagcacga aggagttcat ctactcggag ctgctggcgt
     2821 acctgtactc gtcaggtgat cagaacagtc tgatggagga gtcggcggag caggagcagc
     2881 ggcgggacga gatgctccgc atgtacaacg ctctgaaaga ggctctgaag atcatcgggg
     2941 acgtcagcac atccaccttc accacacctg tgcccgcgcc ggtggacgac acctggatac
     3001 acactgaccc cagtcctccg tctcagcgta agccagctcc gcccactgtc cgccctcctg
     3061 cagtgcgagg ctccaccccc ggtgctccag ccccaccttc tctaaacccc tcccctgcct
     3121 tcggagctcc gcctgttccc tccagaccca ccacattgga gccagctgta gctccgcccc
     3181 cttgtgttcc atctcgccct gcacaggtcc cgcccactgt gctgcccggg atacgcagta
     3241 ggaggcctcc agcggctcca aacaggccca caatcatccg tccgacagaa ccatctctgc
     3301 tggactgaca actttattat acaaagttgg cattataaaa aagcattgct tatcaatttg
     3361 ttgcaacgaa caggtcacta tcagtcaaaa taaaatcatt atttggagct ccatggtagc
     3421 gttaacgcgg ccgcgatatc ccctatagtg agtcgtatta catggtcata gctgtttcct
     3481 ggcagctctg gcccgtgtct caaaatctct gatgttacat tgcacaagat aaaaatatat
     3541 catcatgaac aataaaactg tctgcttaca taaacagtaa tacaaggggt gttatgagcc
     3601 atattcaacg ggaaacgtcg aggccgcgat taaattccaa catggatgct gatttatatg
     3661 ggtataaatg ggctcgcgat aatgtcgggc aatcaggtgc gacaatctat cgcttgtatg
     3721 ggaagcccga tgcgccagag ttgtttctga aacatggcaa aggtagcgtt gccaatgatg
     3781 ttacagatga gatggtcaga ctaaactggc tgacggaatt tatgcctctt ccgaccatca
     3841 agcattttat ccgtactcct gatgatgcat ggttactcac cactgcgatc cccggaaaaa
     3901 cagcattcca ggtattagaa gaatatcctg attcaggtga aaatattgtt gatgcgctgg
     3961 cagtgttcct gcgccggttg cattcgattc ctgtttgtaa ttgtcctttt aacagcgatc
     4021 gcgtatttcg tctcgctcag gcgcaatcac gaatgaataa cggtttggtt gatgcgagtg
     4081 attttgatga cgagcgtaat ggctggcctg ttgaacaagt ctggaaagaa atgcataaac
     4141 ttttgccatt ctcaccggat tcagtcgtca ctcatggtga tttctcactt gataacctta
     4201 tttttgacga ggggaaatta ataggttgta ttgatgttgg acgagtcgga atcgcagacc
     4261 gataccagga tcttgccatc ctatggaact gcctcggtga gttttctcct tcattacaga
     4321 aacggctttt tcaaaaatat ggtattgata atcctgatat gaataaattg cagtttcatt
     4381 tgatgctcga tgagtttttc taatcagaat tggttaattg gttgtaacac tggcagagca
     4441 ttacgctgac ttgacgggac ggcgcaagct catgaccaaa atcccttaac gtgagttacg
     4501 cgtcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     4561 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     4621 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     4681 agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     4741 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     4801 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     4861 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     4921 tgagatacct acagcgtgag cattgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     4981 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     5041 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     5101 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     5161 tacggttcct ggccttttgc tggccttttg ctcacatgtt
//
