LOCUS       sequence-342361-        3637 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Entry clone for tagging MmCavin4 at the C terminus. Parton lab clone
            ADM.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3637)
  AUTHORS   Rae J, Ferguson C, Ariotti N, Webb RI, Cheng HH, Mead JL, Riches JD,
            Hunter DJ, Martel N, Baltos J, Christopoulos A, Bryce NS, Cagigas 
            ML, Fonseka S, Sayre ME, Hardeman EC, Gunning PW, Gambin Y, Hall TE,
            Parton RG
  TITLE     A robust method for particulate detection of a genetic tag for 3D 
            electron microscopy.
  JOURNAL   Elife. 2021 Apr 27;10. pii: 64630. doi: 10.7554/eLife.64630.
  PUBMED    33904409
REFERENCE   2  (bases 1 to 3637)
  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: "Elife."; 
            date: "2021-04-27"; volume: "10. pii: 64630. doi"; pages: " 
            10.7554/eLife.64630"
COMMENT     Sequence Label: pME-MmCavin4
FEATURES             Location/Qualifiers
     source          1..3637
                     /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    569..668
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     regulatory      669..678
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(1762..1861)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1878..1897)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1879..1897)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1902..1918)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1902..1918)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2020..2039)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2031..2840
                     /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(2098..2117)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2987..3575
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3476..3495
                     /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 tcgggcccca aataatgatt ttattttgac tgatagtgac
      601 ctgttcgttg caacaaattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
      661 agcaggctgc caccatggaa cacaacggat cagcttcaaa tgctggtaaa atccaccaga
      721 atcggttgtc aagtgtgacg gaagatgaag accaagacgc agccctcacc attgtgaccg
      781 tgctggacag agtggccagt gtcgtggaca gtgtgcaggc aagccagaag agaattgagg
      841 agagacacag agaaatgggt aacgcgataa agtctgtcca aatagacctg ctgaagctct
      901 cacagtcaca cagcaatacg ggctacgttg tcaacaagct gtttgagaag acccggaaag
      961 tcagcgctca cattaaggat gtcaaggccc gggtagagaa gcaacaggtt cgtgtaacca
     1021 aagtcgaaac caagcaagaa gaaataatga agaaaaacaa attccgagtg gtaatcttcc
     1081 aggaggacat tccctgcccc gcatccctgt ctgttgttaa agacagaagc ctgcccgaga
     1141 accaggagga ggccgaagaa gtcttcgatc ccccaataga gctctcctcg gatgaggagt
     1201 attatgttga agaaagcaga tctgccaggc ttaggaagtc aggcaaagag cacattgacc
     1261 acatcaagaa agccttttcc agagaaaaca tgcagaagac acggcagact cttgacaaga
     1321 aagtgagtgg gattagaact aggatagtca ctcctgagag gagagagagg ctgaggcagt
     1381 cgggagagag gctgaggcag tcaggagaga ggctgaggca gtcgggggaa agatttaaga
     1441 aatcgatttc cagtgccgct ccctcaaagg aagcttttaa gattcggagc cttaggaaag
     1501 cgaaggatcc caaggccgag ggccaggagg tagacagagg gatgggcgtg gacatcatct
     1561 caggtagcct ggctctgggg cccatccatg agttccactc tgatgagttc agtgaaacag
     1621 aaaaggaggt gaccaaagga gggtacagtc cccaagaagg aggggacccc ccaacgcctg
     1681 agcctttgaa ggtgactttt aaaccccagg tgagggtaga ggatgacgag tcactcctgt
     1741 tggagctaaa gcagtcctca tacccagctt tcttgtacaa agttggcatt ataagaaagc
     1801 attgcttatc aatttgttgc aacgaacagg tcactatcag tcaaaataaa atcattattt
     1861 gccatccagc tgatatcccc tatagtgagt cgtattacat ggtcatagct gtttcctggc
     1921 agctctggcc cgtgtctcaa aatctctgat gttacattgc acaagataaa aatatatcat
     1981 catgaacaat aaaactgtct gcttacataa acagtaatac aaggggtgtt atgagccata
     2041 ttcaacggga aacgtcgagg ccgcgattaa attccaacat ggatgctgat ttatatgggt
     2101 ataaatgggc tcgcgataat gtcgggcaat caggtgcgac aatctatcgc ttgtatggga
     2161 agcccgatgc gccagagttg tttctgaaac atggcaaagg tagcgttgcc aatgatgtta
     2221 cagatgagat ggtcagacta aactggctga cggaatttat gcctcttccg accatcaagc
     2281 attttatccg tactcctgat gatgcatggt tactcaccac tgcgatcccc ggaaaaacag
     2341 cattccaggt attagaagaa tatcctgatt caggtgaaaa tattgttgat gcgctggcag
     2401 tgttcctgcg ccggttgcat tcgattcctg tttgtaattg tccttttaac agcgatcgcg
     2461 tatttcgtct cgctcaggcg caatcacgaa tgaataacgg tttggttgat gcgagtgatt
     2521 ttgatgacga gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg cataaacttt
     2581 tgccattctc accggattca gtcgtcactc atggtgattt ctcacttgat aaccttattt
     2641 ttgacgaggg gaaattaata ggttgtattg atgttggacg agtcggaatc gcagaccgat
     2701 accaggatct tgccatccta tggaactgcc tcggtgagtt ttctccttca ttacagaaac
     2761 ggctttttca aaaatatggt attgataatc ctgatatgaa taaattgcag tttcatttga
     2821 tgctcgatga gtttttctaa tcagaattgg ttaattggtt gtaacactgg cagagcatta
     2881 cgctgacttg acgggacggc gcaagctcat gaccaaaatc ccttaacgtg agttacgcgt
     2941 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     3001 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     3061 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     3121 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     3181 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     3241 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     3301 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     3361 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     3421 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     3481 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     3541 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     3601 ggttcctggc cttttgctgg ccttttgctc acatgtt
//
