LOCUS       sequence-222110-        3220 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Multisite gateway vector for 3' tagging with dominant negative 
            RhoAb.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3220)
  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 3220)
  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-RhoAb-DN
FEATURES             Location/Qualifiers
     source          1..3220
                     /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(706..801)
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(838..857)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(839..857)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(862..878)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(862..878)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(980..999)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             991..1800
                     /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(1058..1077)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      1947..2535
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2436..2455
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2689..2706
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2865..2892)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(2984..3070)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3119..3141
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3133..3150
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3134..3150
                     /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 gtggctgcag caattcgcaa gaagctggtg attgtaggag atgtagcatg cgggaaaaca
      181 tgtttgctta ttgtgttcag taaagaccag tttcctgaag tctacgtacc aacagtgttt
      241 gaaaactatg ttgccgacat tgaagttgac agcaaacagg tggaactcgc tctgtgggat
      301 actgcagggc aggaggacta cgacaggctt cgtcctcttt catacccaga caccgatgtc
      361 attcttatgt gtttctctat tgacagtcct gacagtttag agaatattcc ggagaagtgg
      421 acgcctgagg tcaaacactt ttgtccaaac gttccaatca tccttgtggg aaacaaaaaa
      481 gacctgcgca atgatgaaca cacacgcagg gaactgacca agatgaagca ggagccagtt
      541 aaagcagagg aagggagaga catggccaat cgaattggag catttggtta tatggaatgt
      601 tctgccaaaa caaaagacgg tgttcgggag gtgtttgaaa tggccactag agcggcgctg
      661 caggcacgca ggggaaagaa gagcaacaaa tgctgtctgc tgtgacaact ttattataca
      721 aagttggcat tataaaaaag cattgcttat caatttgttg caacgaacag gtcactatca
      781 gtcaaaataa aatcattatt tggagctcca tggtagcgtt aacgcggccg cgatatcccc
      841 tatagtgagt cgtattacat ggtcatagct gtttcctggc agctctggcc cgtgtctcaa
      901 aatctctgat gttacattgc acaagataaa aatatatcat catgaacaat aaaactgtct
      961 gcttacataa acagtaatac aaggggtgtt atgagccata ttcaacggga aacgtcgagg
     1021 ccgcgattaa attccaacat ggatgctgat ttatatgggt ataaatgggc tcgcgataat
     1081 gtcgggcaat caggtgcgac aatctatcgc ttgtatggga agcccgatgc gccagagttg
     1141 tttctgaaac atggcaaagg tagcgttgcc aatgatgtta cagatgagat ggtcagacta
     1201 aactggctga cggaatttat gcctcttccg accatcaagc attttatccg tactcctgat
     1261 gatgcatggt tactcaccac tgcgatcccc ggaaaaacag cattccaggt attagaagaa
     1321 tatcctgatt caggtgaaaa tattgttgat gcgctggcag tgttcctgcg ccggttgcat
     1381 tcgattcctg tttgtaattg tccttttaac agcgatcgcg tatttcgtct cgctcaggcg
     1441 caatcacgaa tgaataacgg tttggttgat gcgagtgatt ttgatgacga gcgtaatggc
     1501 tggcctgttg aacaagtctg gaaagaaatg cataaacttt tgccattctc accggattca
     1561 gtcgtcactc atggtgattt ctcacttgat aaccttattt ttgacgaggg gaaattaata
     1621 ggttgtattg atgttggacg agtcggaatc gcagaccgat accaggatct tgccatccta
     1681 tggaactgcc tcggtgagtt ttctccttca ttacagaaac ggctttttca aaaatatggt
     1741 attgataatc ctgatatgaa taaattgcag tttcatttga tgctcgatga gtttttctaa
     1801 tcagaattgg ttaattggtt gtaacactgg cagagcatta cgctgacttg acgggacggc
     1861 gcaagctcat gaccaaaatc ccttaacgtg agttacgcgt cgttccactg agcgtcagac
     1921 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     1981 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     2041 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     2101 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     2161 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     2221 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     2281 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     2341 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     2401 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     2461 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     2521 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     2581 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     2641 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
     2701 agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt
     2761 cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca
     2821 attaatacgc gtaccgctag ccaggaagag tttgtagaaa cgcaaaaagg ccatccgtca
     2881 ggatggcctt ctgcttagtt tgatgcctgg cagtttatgg cgggcgtcct gcccgccacc
     2941 ctccgggccg ttgcttcaca acgttcaaat ccgctcccgg cggatttgtc ctactcagga
     3001 gagcgttcac cgacaaacaa cagataaaac gaaaggccca gtcttccgac tgagcctttc
     3061 gttttatttg atgcctggca gttccctact ctcgcgttaa cgctagcatg gatgttttcc
     3121 cagtcacgac gttgtaaaac gacggccagt cttaagctcg ggccctgcag ctctagagct
     3181 cgaattctac aggtcactaa taccatctaa gtagttggtt
//
