LOCUS       sequence-206501-        4240 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Multisite gateway vector for 3' tagging with EHD1a.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4240)
  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 4240)
  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-EHD1a
FEATURES             Location/Qualifiers
     source          1..4240
                     /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(2349..2444)
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2481..2500)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2482..2500)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2505..2521)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2505..2521)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2623..2642)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2634..3443
                     /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(2701..2720)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3590..4178
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4079..4098
                     /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 aaagtggctt tcagctggtc gaatagagat ggtaagaaag
      781 acccagaact tttccaaaac gtgtccgatg gactgaagcg cctttaccgc accaaactct
      841 tccctctgga ggacacctat catttccacg acttccattc accggcgctc gaagatgcgg
      901 attttgacaa caagcccatg gtgctgctgg tgggacagta ctcgaccggg aagaccacct
      961 tcatcaggca tttaatggag caggatttcc ccgggatgag gataggccca gagccgacga
     1021 cggattcctt cattgcggtt atgcacggtg agcaagaagg cgtcatacct ggtaatgcgc
     1081 tggtggtgga tccgaaaaag cctttccgaa aactcaacgc atttggcaac gctttcctca
     1141 acaggtttat gtgtgcacag ttaaacaacc cggtacttga gagcatcagt atcatcgata
     1201 ctccaggcat cctgtcaggg gagaaacaga ggatcagcag aggctatgac tttgctgctg
     1261 ttctggagtg gtttgcggag cgcgtggacc gaatcatcct gctcttcgac gcccacaagc
     1321 tggacatttc ggatgagttt tcagaggtga tcaaggccct gaagaaccac gaggacaaga
     1381 tgcgcgtggt gctgaataaa gcagatcaga tcagcacgca gcagctcatg agggtttatg
     1441 gagccctcat gtggtcattg ggcaagatca taaacacccc tgaggtggtg cgggtttata
     1501 tcggctcctt ctgggctcag ccgctgctca tcgctgaaaa ccggaagctg tttgaagcgg
     1561 aggagcagga tcttttccag gatattcaga gtctgccgag aaacgcagct ctgaggaaac
     1621 tcaacgatct gatcaaaaga gcacgtcttg ctaaggtaca tgcttacatt atcagtgctc
     1681 tgaagaaaga gatgccgagc gtgtttggca aagactcaaa gaagaaagag cttattaaca
     1741 acttgggcgc tatatatgaa aagatccaga aagagcacaa catttcacca ggagactttc
     1801 caaaccttaa gaaaatgcag gagttgctat ctgttcagga cttcaccaaa ttccctgcca
     1861 tgaagcccaa acttctagag tcggtggaag acatgctagg taacgacatc gctaagctaa
     1921 tggcactggt gcgtaaagag gaggccacca tgcccagcca atcagtgaag ggcggtgctt
     1981 ttgagggaat ctcaaacggc ccgttcggtc atgggtacgg agaaggcgcc ggcgaaggca
     2041 tcgacgagct ggagtggatc gtgggccgtg acaaaccatc ctacgatgaa attttctaca
     2101 ccctctctcc gatcaacggg aagatctccg gagccacagc taagaaggag atgacgaagt
     2161 ccaagctgcc gaacacagtg ctggggaaaa tctggactct ggcagatgtg gataaagatg
     2221 gatacctgga tgacgaagag tttgctctgg ccaatcacct gatcaaggta aagcttgagg
     2281 gcttcgagct cccggctaaa cttcccaatc atcttgtgcc tccgtccaaa cggcaagagc
     2341 acaattgaca actttattat acaaagttgg cattataaaa aagcattgct tatcaatttg
     2401 ttgcaacgaa caggtcacta tcagtcaaaa taaaatcatt atttggagct ccatggtagc
     2461 gttaacgcgg ccgcgatatc ccctatagtg agtcgtatta catggtcata gctgtttcct
     2521 ggcagctctg gcccgtgtct caaaatctct gatgttacat tgcacaagat aaaaatatat
     2581 catcatgaac aataaaactg tctgcttaca taaacagtaa tacaaggggt gttatgagcc
     2641 atattcaacg ggaaacgtcg aggccgcgat taaattccaa catggatgct gatttatatg
     2701 ggtataaatg ggctcgcgat aatgtcgggc aatcaggtgc gacaatctat cgcttgtatg
     2761 ggaagcccga tgcgccagag ttgtttctga aacatggcaa aggtagcgtt gccaatgatg
     2821 ttacagatga gatggtcaga ctaaactggc tgacggaatt tatgcctctt ccgaccatca
     2881 agcattttat ccgtactcct gatgatgcat ggttactcac cactgcgatc cccggaaaaa
     2941 cagcattcca ggtattagaa gaatatcctg attcaggtga aaatattgtt gatgcgctgg
     3001 cagtgttcct gcgccggttg cattcgattc ctgtttgtaa ttgtcctttt aacagcgatc
     3061 gcgtatttcg tctcgctcag gcgcaatcac gaatgaataa cggtttggtt gatgcgagtg
     3121 attttgatga cgagcgtaat ggctggcctg ttgaacaagt ctggaaagaa atgcataaac
     3181 ttttgccatt ctcaccggat tcagtcgtca ctcatggtga tttctcactt gataacctta
     3241 tttttgacga ggggaaatta ataggttgta ttgatgttgg acgagtcgga atcgcagacc
     3301 gataccagga tcttgccatc ctatggaact gcctcggtga gttttctcct tcattacaga
     3361 aacggctttt tcaaaaatat ggtattgata atcctgatat gaataaattg cagtttcatt
     3421 tgatgctcga tgagtttttc taatcagaat tggttaattg gttgtaacac tggcagagca
     3481 ttacgctgac ttgacgggac ggcgcaagct catgaccaaa atcccttaac gtgagttacg
     3541 cgtcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     3601 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     3661 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     3721 agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     3781 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     3841 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     3901 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     3961 tgagatacct acagcgtgag cattgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     4021 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     4081 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     4141 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     4201 tacggttcct ggccttttgc tggccttttg ctcacatgtt
//
