LOCUS       sequence-206503-        4237 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Multisite gateway vector for 3' tagging with EHD1b.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4237)
  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 4237)
  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-EHD1b
FEATURES             Location/Qualifiers
     source          1..4237
                     /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(2346..2441)
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2478..2497)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2479..2497)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2502..2518)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2502..2518)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2620..2639)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2631..3440
                     /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(2698..2717)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3587..4175
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4076..4095
                     /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 tcagttggtc taacaaagat ggaaaaaaga
      781 aagacccgga gttatttcaa acggtttcgg acgggctaaa gagactgtac aggaccaaac
      841 ttttcccctt agaggacgca taccgctttc acgacttcca ctctccggct ctggaagatg
      901 ctgatttcga taacaagccc atggtacttt tggtgggcca atactctaca gggaaaacca
      961 cttttattcg gcatctcatg gagcaggatt tccctggtat gcgcataggg cccgaaccga
     1021 ccaccgattc attcatagcg gttatgcacg gggaacaaga aggtctcatc cccggcaatg
     1081 ctttagtggt tgaccccaag aaacccttca gaaaactcaa cgcctttgga aacgcctttc
     1141 tgaacaggtt catatgtgct cagatgccaa acccagtgct ggacagcatc agcatcatag
     1201 acactcctgg aattctgtcc ggagagaaac aacggatcag cagaggttat gattttgctg
     1261 cggtgcttga atggtttgcc gaacgggtgg accgaatcat tcttctcttt gacgcacaca
     1321 agttggacat ctcggatgag ttctccgagg tgatccgagc tctgaaaaac catgaggaca
     1381 aaatgcgggt ggtgcttaat aaagctgatc agatcgggac ccagcagctt atgagggtgt
     1441 atggcgccct catgtggtca cttgggaaaa ttgtcaacac tccagaggtc atacgtgtgt
     1501 atattggctc tttctgggct cagccgctcc tggttgcaga taacaggaaa ttgtttgagg
     1561 cagaggaaca agaccttttc cgtgacatcc agggacttcc acgtaatgca gcactgcgca
     1621 aactgaatga ccttatcaaa cgggcccgtc tagccaaagt tcaagcatac atcatcagct
     1681 ctttgaagaa agaaatgcca aatgtgtttg gaaaagagaa caagaaaaag gagctcattg
     1741 ctaacctggg tgaaatctat gcaaagattg aaaaagaaca tcagatctcc cctggtgatt
     1801 ttcctaaact cagtaaaatg caggaagtcc tcgctggaca ggacttcact aagtttcaag
     1861 gattgaagcc caagcttttg gaatcagtgg aagacatgct ggccaacgac attgccaaac
     1921 tcatgaccct cgtccggcag gaggaggcct ccatgccaag ccaggctgtc caaggtggag
     1981 cctttgaagg caccatgaat ggccccttcg ggcacggcta cggagaaggt gccggtgaag
     2041 gcattgatga gctggagtgg gtagtggcca aagacaaacc gacatacgac gagatcttct
     2101 acactttgtc acccgtcaat gggaaggtgt ctggtgccat ggccaagaag gagatggtta
     2161 agtccaagct gcccaataat gttctaggaa agatctggaa gctcgctgac gtggataagg
     2221 atggatacct ggatgacgag gagttcgccc tggccaacca cctcatcaag gttaaactgg
     2281 aaggtcacga gttgccagct gatcttccag atcaccttgt gccaccttca aaacgagaac
     2341 agtaacaact ttattataca aagttggcat tataaaaaag cattgcttat caatttgttg
     2401 caacgaacag gtcactatca gtcaaaataa aatcattatt tggagctcca tggtagcgtt
     2461 aacgcggccg cgatatcccc tatagtgagt cgtattacat ggtcatagct gtttcctggc
     2521 agctctggcc cgtgtctcaa aatctctgat gttacattgc acaagataaa aatatatcat
     2581 catgaacaat aaaactgtct gcttacataa acagtaatac aaggggtgtt atgagccata
     2641 ttcaacggga aacgtcgagg ccgcgattaa attccaacat ggatgctgat ttatatgggt
     2701 ataaatgggc tcgcgataat gtcgggcaat caggtgcgac aatctatcgc ttgtatggga
     2761 agcccgatgc gccagagttg tttctgaaac atggcaaagg tagcgttgcc aatgatgtta
     2821 cagatgagat ggtcagacta aactggctga cggaatttat gcctcttccg accatcaagc
     2881 attttatccg tactcctgat gatgcatggt tactcaccac tgcgatcccc ggaaaaacag
     2941 cattccaggt attagaagaa tatcctgatt caggtgaaaa tattgttgat gcgctggcag
     3001 tgttcctgcg ccggttgcat tcgattcctg tttgtaattg tccttttaac agcgatcgcg
     3061 tatttcgtct cgctcaggcg caatcacgaa tgaataacgg tttggttgat gcgagtgatt
     3121 ttgatgacga gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg cataaacttt
     3181 tgccattctc accggattca gtcgtcactc atggtgattt ctcacttgat aaccttattt
     3241 ttgacgaggg gaaattaata ggttgtattg atgttggacg agtcggaatc gcagaccgat
     3301 accaggatct tgccatccta tggaactgcc tcggtgagtt ttctccttca ttacagaaac
     3361 ggctttttca aaaatatggt attgataatc ctgatatgaa taaattgcag tttcatttga
     3421 tgctcgatga gtttttctaa tcagaattgg ttaattggtt gtaacactgg cagagcatta
     3481 cgctgacttg acgggacggc gcaagctcat gaccaaaatc ccttaacgtg agttacgcgt
     3541 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     3601 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     3661 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     3721 taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     3781 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     3841 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     3901 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     3961 gatacctaca gcgtgagcat tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     4021 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     4081 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     4141 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     4201 ggttcctggc cttttgctgg ccttttgctc acatgtt
//
