LOCUS       sequence-222045-        4270 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Multisite gateway vector for 3' tagging with EHD2b.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4270)
  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 4270)
  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-EHD2b
FEATURES             Location/Qualifiers
     source          1..4270
                     /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(1756..1851)
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1888..1907)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1889..1907)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1912..1928)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1912..1928)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2030..2049)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2041..2850
                     /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(2108..2127)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2997..3585
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3486..3505
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3739..3756
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3915..3942)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4034..4120)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     4169..4191
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4183..4200
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4184..4200
                     /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 gtggtgtcac gctggggacg taagaatgtc aagaaggccc ccgaggtgat caggacagtg
      181 accgagggcc tgaagtcact ctaccgaaaa aagctcctgc ctctggagca gtattacggc
      241 ttccatgact tccactcacc gagtctggag gacgccgact ttgacaacaa acccatggtg
      301 ctggtcgttg ggcagtactc cacgggaaag accaccttca tcaagtatct gcttgaacag
      361 gatgtccctg ggagccgcat agggcctgag cccaccaccg actgcttcac cgctatcatg
      421 cacggagacg tagaggggct cattcccgga aatgccctca tcgtggatcc caacaagccc
      481 ttccgcaaac tcaacccctt cggaaacacc tttctcaaca gattccagtg cgctcagctg
      541 cccaaccagg tcctggagag catcagcatt attgacaccc caggaatcct gtctggggcc
      601 aaacagagag tgagtcgagg atatgatttc ccagcagttc tccgctggtt tgctgagcgt
      661 gtggaccgaa tcatccttct ctttgatgcc cacaagttag agatctcaga tgaattctca
      721 gaggctatcg gagctctcaa aggcaatgaa gacaaactcc gtgttgtgct gaacaaggct
      781 gatatggttg atacccagca gcttatgcgg gtgtacggag ctttaatgtg gtccctgggg
      841 aaagtgtttg gtactcctga ggttctgaga gtgtacatcg gttccttctg gtctgaacct
      901 ctcatggtga ccgacaaccg caagctgttc gaattggagg aggaagatct gtttgctgat
      961 atccagaatc tgccacgtaa tgcagcccta cgcaaactta atgacttggt caagagggcc
     1021 cgtcttgtga gggtacatgc ccatattatc agttacttga aacaagagat gccatcggtc
     1081 ttcagaaaag acaataaaaa gaagaacctg atctaccagc tcccggtcat cttctccaag
     1141 atccaactac aacatcacat ctctcctgga gacttccctg actgtgccaa gatgcaggag
     1201 caactaatgg cacatgactt caccaagttt aaagccctga agcccaacct tatgaatatg
     1261 ctggatgagc tgttgtccag tgacattgcc aaactgatgc cccttcttag gcaggaagaa
     1321 atagaggctg gggttcaacc aggtgtccaa ggtggagctt tcttgggaac ccgtgccgga
     1381 ccctttgtgg aaggtgaccc ctttggagag gttgcgaatg aaaatggcga agtgggtgaa
     1441 gaagatgagg aatgggtggt gacaaaagac aagcccaagt atgatgagat cttctacaac
     1501 ctcgcaccga atgaaggcaa actgagtggc accaaagcca aagattggat ggtgagcaca
     1561 aggctcccca actctgtttt gggtcgtatc tggaagcttt cggatgtgga tcgtgacggc
     1621 atgctggatg atgaggagtt tgccctggcc agccatctga ttgaagtgaa actggatggt
     1681 catgggctgc cccctgagct ccctgcccgt ctcgtgccac catccaagcg ccggcaaaaa
     1741 ggctctgatg cttagcaact ttattataca aagttggcat tataaaaaag cattgcttat
     1801 caatttgttg caacgaacag gtcactatca gtcaaaataa aatcattatt tggagctcca
     1861 tggtagcgtt aacgcggccg cgatatcccc tatagtgagt cgtattacat ggtcatagct
     1921 gtttcctggc agctctggcc cgtgtctcaa aatctctgat gttacattgc acaagataaa
     1981 aatatatcat catgaacaat aaaactgtct gcttacataa acagtaatac aaggggtgtt
     2041 atgagccata ttcaacggga aacgtcgagg ccgcgattaa attccaacat ggatgctgat
     2101 ttatatgggt ataaatgggc tcgcgataat gtcgggcaat caggtgcgac aatctatcgc
     2161 ttgtatggga agcccgatgc gccagagttg tttctgaaac atggcaaagg tagcgttgcc
     2221 aatgatgtta cagatgagat ggtcagacta aactggctga cggaatttat gcctcttccg
     2281 accatcaagc attttatccg tactcctgat gatgcatggt tactcaccac tgcgatcccc
     2341 ggaaaaacag cattccaggt attagaagaa tatcctgatt caggtgaaaa tattgttgat
     2401 gcgctggcag tgttcctgcg ccggttgcat tcgattcctg tttgtaattg tccttttaac
     2461 agcgatcgcg tatttcgtct cgctcaggcg caatcacgaa tgaataacgg tttggttgat
     2521 gcgagtgatt ttgatgacga gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg
     2581 cataaacttt tgccattctc accggattca gtcgtcactc atggtgattt ctcacttgat
     2641 aaccttattt ttgacgaggg gaaattaata ggttgtattg atgttggacg agtcggaatc
     2701 gcagaccgat accaggatct tgccatccta tggaactgcc tcggtgagtt ttctccttca
     2761 ttacagaaac ggctttttca aaaatatggt attgataatc ctgatatgaa taaattgcag
     2821 tttcatttga tgctcgatga gtttttctaa tcagaattgg ttaattggtt gtaacactgg
     2881 cagagcatta cgctgacttg acgggacggc gcaagctcat gaccaaaatc ccttaacgtg
     2941 agttacgcgt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     3001 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     3061 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     3121 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     3181 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     3241 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     3301 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     3361 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     3421 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     3481 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     3541 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     3601 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     3661 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     3721 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc
     3781 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc
     3841 gactggaaag cgggcagtga gcgcaacgca attaatacgc gtaccgctag ccaggaagag
     3901 tttgtagaaa cgcaaaaagg ccatccgtca ggatggcctt ctgcttagtt tgatgcctgg
     3961 cagtttatgg cgggcgtcct gcccgccacc ctccgggccg ttgcttcaca acgttcaaat
     4021 ccgctcccgg cggatttgtc ctactcagga gagcgttcac cgacaaacaa cagataaaac
     4081 gaaaggccca gtcttccgac tgagcctttc gttttatttg atgcctggca gttccctact
     4141 ctcgcgttaa cgctagcatg gatgttttcc cagtcacgac gttgtaaaac gacggccagt
     4201 cttaagctcg ggccctgcag ctctagagct cgaattctac aggtcactaa taccatctaa
     4261 gtagttggtt
//
