LOCUS       sequence-222044-        4279 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Multisite gateway vector for 3' tagging with EHD2a.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4279)
  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 4279)
  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-EHD2a
FEATURES             Location/Qualifiers
     source          1..4279
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(1..125)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(1765..1860)
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1897..1916)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1898..1916)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1921..1937)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1921..1937)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2039..2058)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2050..2859
                     /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(2117..2136)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3006..3594
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3495..3514
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3748..3765
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3924..3951)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4043..4129)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     4178..4200
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4192..4209
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4193..4209
                     /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 gtggtgtctg ccaaaggaat gaggaacagg ggtggtaaat gtctgaaggc ggttcataca
      181 gtaaccgagg ctttgaaaat cctctatcgg caaaaactgc tcccactgga gaagtactat
      241 ggtttctccg atttccactc atcaagcctt gaagatgcag actttgacaa caaacccatg
      301 atactggtgt tgggccagtg ctctacaggc aagaccactt ttatcaggta tcttcttgaa
      361 caggagtatc ctggaagcag agttggccca gaaccgacaa ccgacagttt cacagcaata
      421 atgcatggag agatggaggg aattattcat ggcaatgctt taactgttga tcccaataag
      481 cctttccgca aactcagccc atttggaaca actttcctaa atcgattcca gtgtgtacat
      541 ctgcccaatc gggtcctaga gagcatcagc atcatcgaca caccagggat cctgtctgtg
      601 gccaagcgca aaatggacag aggttatgac tttccttcag ttatccgctg gtttgcggag
      661 cacgtagacc ggatcatcct cctgtgtgat gcacacaagc tggagatttc tgatgagttt
      721 tcccgcacta tactggcatt acgaggcaat gaggacaagc tccgagtggt gctcaacaag
      781 gccgacatgg tggactcgca ggagctgatg cgggtctacg gcgccctcat gtggtctctc
      841 gggaaggtct tctgcacccc ggaggtcctg agggtctaca tcggctcctt ctggaccgct
      901 ccgatgcgaa tcacagacaa ccgcaaactg tttgagcgag aggagggaga tctttttgca
      961 gaccttcaga acctgccaag aaattctgcc ctgagaaaac tcaatgatct ggtgaaacga
     1021 gcacgccttg taagggctca tgcttatatc atcagctgtc taaaagacga gatgcctgtt
     1081 gttttctgca gaggaaagaa gcggcgagat ctcatctatg accttcccac catctattcc
     1141 agaatccaac agcgctatca gatttcacca ggagattttc cagactgtgc caagatgcag
     1201 gagcgactca tgagccacga cttcaagttt aaaggcctca agcccaagct cttagctggg
     1261 ttggatgagc tcctaaacac cgacattgct aagctgatgc ctcttctacg ccaagaggaa
     1321 gaagaagcag ctgatcaact tttaatacaa ggaggacctt tagggtctca caggggcccg
     1381 tttattgtgg ggaacccatt tcatcactct ggatccaatg gagaaatcga tcaggcgtca
     1441 aatgatgaat gggtggtgtc aaaagataaa gcaaagtacg acgagatctt ttacaacctc
     1501 tcgcctcatg agggaaagct gactgggact aaagtgaagg agtggatgtt gcgcactcat
     1561 ctgcccagct cggtcctggg acgaatctgg aagctggcag atgtggattg tgatggtaag
     1621 ttggacgatg aggagtttgc tttggccggt catctcattg aggtaaagtt ggaaggtttt
     1681 gggcttcctc atgagctgcc cacacacctg ctgccaccgt ctaaaagaca ccgcaagaac
     1741 tctgagatta ccgacgatga ttagcaactt tattatacaa agttggcatt ataaaaaagc
     1801 attgcttatc aatttgttgc aacgaacagg tcactatcag tcaaaataaa atcattattt
     1861 ggagctccat ggtagcgtta acgcggccgc gatatcccct atagtgagtc gtattacatg
     1921 gtcatagctg tttcctggca gctctggccc gtgtctcaaa atctctgatg ttacattgca
     1981 caagataaaa atatatcatc atgaacaata aaactgtctg cttacataaa cagtaataca
     2041 aggggtgtta tgagccatat tcaacgggaa acgtcgaggc cgcgattaaa ttccaacatg
     2101 gatgctgatt tatatgggta taaatgggct cgcgataatg tcgggcaatc aggtgcgaca
     2161 atctatcgct tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca tggcaaaggt
     2221 agcgttgcca atgatgttac agatgagatg gtcagactaa actggctgac ggaatttatg
     2281 cctcttccga ccatcaagca ttttatccgt actcctgatg atgcatggtt actcaccact
     2341 gcgatccccg gaaaaacagc attccaggta ttagaagaat atcctgattc aggtgaaaat
     2401 attgttgatg cgctggcagt gttcctgcgc cggttgcatt cgattcctgt ttgtaattgt
     2461 ccttttaaca gcgatcgcgt atttcgtctc gctcaggcgc aatcacgaat gaataacggt
     2521 ttggttgatg cgagtgattt tgatgacgag cgtaatggct ggcctgttga acaagtctgg
     2581 aaagaaatgc ataaactttt gccattctca ccggattcag tcgtcactca tggtgatttc
     2641 tcacttgata accttatttt tgacgagggg aaattaatag gttgtattga tgttggacga
     2701 gtcggaatcg cagaccgata ccaggatctt gccatcctat ggaactgcct cggtgagttt
     2761 tctccttcat tacagaaacg gctttttcaa aaatatggta ttgataatcc tgatatgaat
     2821 aaattgcagt ttcatttgat gctcgatgag tttttctaat cagaattggt taattggttg
     2881 taacactggc agagcattac gctgacttga cgggacggcg caagctcatg accaaaatcc
     2941 cttaacgtga gttacgcgtc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     3001 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     3061 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     3121 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     3181 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     3241 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     3301 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     3361 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     3421 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     3481 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     3541 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     3601 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     3661 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     3721 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     3781 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac
     3841 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatacgcg taccgctagc
     3901 caggaagagt ttgtagaaac gcaaaaaggc catccgtcag gatggccttc tgcttagttt
     3961 gatgcctggc agtttatggc gggcgtcctg cccgccaccc tccgggccgt tgcttcacaa
     4021 cgttcaaatc cgctcccggc ggatttgtcc tactcaggag agcgttcacc gacaaacaac
     4081 agataaaacg aaaggcccag tcttccgact gagcctttcg ttttatttga tgcctggcag
     4141 ttccctactc tcgcgttaac gctagcatgg atgttttccc agtcacgacg ttgtaaaacg
     4201 acggccagtc ttaagctcgg gccctgcagc tctagagctc gaattctaca ggtcactaat
     4261 accatctaag tagttggtt
//
