LOCUS       sequence-222039-        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"
     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(1726..1821)
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1858..1877)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1859..1877)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1882..1898)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1882..1898)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2000..2019)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2011..2820
                     /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(2078..2097)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2967..3555
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3456..3475
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3709..3726
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3885..3912)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4004..4090)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     4139..4161
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4153..4170
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4154..4170
                     /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 gtggctttca gctggtcgaa tagagatggt aagaaagacc cagaactttt ccaaaacgtg
      181 tccgatggac tgaagcgcct ttaccgcacc aaactcttcc ctctggagga cacctatcat
      241 ttccacgact tccattcacc ggcgctcgaa gatgcggatt ttgacaacaa gcccatggtg
      301 ctgctggtgg gacagtactc gaccgggaag accaccttca tcaggcattt aatggagcag
      361 gatttccccg ggatgaggat aggcccagag ccgacgacgg attccttcat tgcggttatg
      421 cacggtgagc aagaaggcgt catacctggt aatgcgctgg tggtggatcc gaaaaagcct
      481 ttccgaaaac tcaacgcatt tggcaacgct ttcctcaaca ggtttatgtg tgcacagtta
      541 aacaacccgg tacttgagag catcagtatc atcgatactc caggcatcct gtcaggggag
      601 aaacagagga tcagcagagg ctatgacttt gctgctgttc tggagtggtt tgcggagcgc
      661 gtggaccgaa tcatcctgct cttcgacgcc cacaagctgg acatttcgga tgagttttca
      721 gaggtgatca aggccctgaa gaaccacgag gacaagatgc gcgtggtgct gaataaagca
      781 gatcagatca gcacgcagca gctcatgagg gtttatggag ccctcatgtg gtcattgggc
      841 aagatcataa acacccctga ggtggtgcgg gtttatatcg gctccttctg ggctcagccg
      901 ctgctcatcg ctgaaaaccg gaagctgttt gaagcggagg agcaggatct tttccaggat
      961 attcagagtc tgccgagaaa cgcagctctg aggaaactca acgatctgat caaaagagca
     1021 cgtcttgcta aggtacatgc ttacattatc agtgctctga agaaagagat gccgagcgtg
     1081 tttggcaaag actcaaagaa gaaagagctt attaacaact tgggcgctat atatgaaaag
     1141 atccagaaag agcacaacat ttcaccagga gactttccaa accttaagaa aatgcaggag
     1201 ttgctatctg ttcaggactt caccaaattc cctgccatga agcccaaact tctagagtcg
     1261 gtggaagaca tgctaggtaa cgacatcgct aagctaatgg cactggtgcg taaagaggag
     1321 gccaccatgc ccagccaatc agtgaagggc ggtgcttttg agggaatctc aaacggcccg
     1381 ttcggtcatg ggtacggaga aggcgccggc gaaggcatcg acgagctgga gtggatcgtg
     1441 ggccgtgaca aaccatccta cgatgaaatt ttctacaccc tctctccgat caacgggaag
     1501 atctccggag ccacagctaa gaaggagatg acgaagtcca agctgccgaa cacagtgctg
     1561 gggaaaatct ggactctggc agatgtggat aaagatggat acctggatga cgaagagttt
     1621 gctctggcca atcacctgat caaggtaaag cttgagggct tcgagctccc ggctaaactt
     1681 cccaatcatc ttgtgcctcc gtccaaacgg caagagcaca attgacaact ttattataca
     1741 aagttggcat tataaaaaag cattgcttat caatttgttg caacgaacag gtcactatca
     1801 gtcaaaataa aatcattatt tggagctcca tggtagcgtt aacgcggccg cgatatcccc
     1861 tatagtgagt cgtattacat ggtcatagct gtttcctggc agctctggcc cgtgtctcaa
     1921 aatctctgat gttacattgc acaagataaa aatatatcat catgaacaat aaaactgtct
     1981 gcttacataa acagtaatac aaggggtgtt atgagccata ttcaacggga aacgtcgagg
     2041 ccgcgattaa attccaacat ggatgctgat ttatatgggt ataaatgggc tcgcgataat
     2101 gtcgggcaat caggtgcgac aatctatcgc ttgtatggga agcccgatgc gccagagttg
     2161 tttctgaaac atggcaaagg tagcgttgcc aatgatgtta cagatgagat ggtcagacta
     2221 aactggctga cggaatttat gcctcttccg accatcaagc attttatccg tactcctgat
     2281 gatgcatggt tactcaccac tgcgatcccc ggaaaaacag cattccaggt attagaagaa
     2341 tatcctgatt caggtgaaaa tattgttgat gcgctggcag tgttcctgcg ccggttgcat
     2401 tcgattcctg tttgtaattg tccttttaac agcgatcgcg tatttcgtct cgctcaggcg
     2461 caatcacgaa tgaataacgg tttggttgat gcgagtgatt ttgatgacga gcgtaatggc
     2521 tggcctgttg aacaagtctg gaaagaaatg cataaacttt tgccattctc accggattca
     2581 gtcgtcactc atggtgattt ctcacttgat aaccttattt ttgacgaggg gaaattaata
     2641 ggttgtattg atgttggacg agtcggaatc gcagaccgat accaggatct tgccatccta
     2701 tggaactgcc tcggtgagtt ttctccttca ttacagaaac ggctttttca aaaatatggt
     2761 attgataatc ctgatatgaa taaattgcag tttcatttga tgctcgatga gtttttctaa
     2821 tcagaattgg ttaattggtt gtaacactgg cagagcatta cgctgacttg acgggacggc
     2881 gcaagctcat gaccaaaatc ccttaacgtg agttacgcgt cgttccactg agcgtcagac
     2941 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     3001 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     3061 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     3121 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     3181 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     3241 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     3301 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     3361 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     3421 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     3481 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     3541 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     3601 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     3661 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
     3721 agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt
     3781 cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca
     3841 attaatacgc gtaccgctag ccaggaagag tttgtagaaa cgcaaaaagg ccatccgtca
     3901 ggatggcctt ctgcttagtt tgatgcctgg cagtttatgg cgggcgtcct gcccgccacc
     3961 ctccgggccg ttgcttcaca acgttcaaat ccgctcccgg cggatttgtc ctactcagga
     4021 gagcgttcac cgacaaacaa cagataaaac gaaaggccca gtcttccgac tgagcctttc
     4081 gttttatttg atgcctggca gttccctact ctcgcgttaa cgctagcatg gatgttttcc
     4141 cagtcacgac gttgtaaaac gacggccagt cttaagctcg ggccctgcag ctctagagct
     4201 cgaattctac aggtcactaa taccatctaa gtagttggtt
//
