LOCUS       sequence-127759-        3466 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Multisite gateway entry clone for adding C-terminal fusions of human
            2PH-PLC-delta.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3466)
  AUTHORS   Ariotti N, Hall TE, Rae J, Ferguson C, McMahon KA, Martel N, Webb 
            RE, Webb RI, Teasdale RD, Parton RG
  TITLE     Modular Detection of GFP-Labeled Proteins for Rapid Screening by 
            Electron Microscopy in Cells and Organisms.
  JOURNAL   Dev Cell. 2015 Nov 23;35(4):513-25. doi: 
            10.1016/j.devcel.2015.10.016. Epub 2015 Nov 12.
  PUBMED    26585296
REFERENCE   2  (bases 1 to 3466)
  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"; doi: 
            "10.1016/j.devcel.2015.10.016"; journalName: "Dev Cell"; date: 
            "2015-11-23- 23"; volume: "35"; issue: "4"; pages: "513-25"
COMMENT     Sequence Label: pME-2PH-PLCd
FEATURES             Location/Qualifiers
     source          1..3466
                     /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      672..681
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(1591..1690)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1707..1726)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1708..1726)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1731..1747)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1731..1747)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1849..1868)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1860..2669
                     /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(1927..1946)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2816..3404
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3305..3324
                     /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 caacacattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
      661 agcaggctgc cgccaccatg gactcgggcc gggacttcct gaccctgcac ggcctacagg
      721 atgatgagga tctacaggcg ctgctgaagg gcagccagct cctgaaggtg aagtccagct
      781 catggaggag agagcgcttc tacaagttgc aggaggactg caagaccatc tggcaggagt
      841 cccgcaaggt catgcggacc ccggagtccc agctgttctc catcgaggac attcaggagg
      901 tgcgaatggg gcaccgcacg gagggtctgg agaagttcgc ccgtgatgtg cccgaggacc
      961 gctgcttctc cattgtcttc aaggaccagc gcaatacact agacctcatc gccccatcgc
     1021 cagctgatgc ccagcactgg gtgctggggc tgcacaaaat tatccatcac agcggatcta
     1081 tggaccagcg tcagaagcta cagcactgga ttcactcctg cttgcgaaaa gctgacaaaa
     1141 acaaggacaa caagataggc ggcagcggcg gcagcggcgg cgatagcggt agagattttc
     1201 tgactctgca tggactgcag gacgacgagg acctgcaggc cctgctgaag ggcagccagc
     1261 tgctgaaggt gaagagcagc agctggcgcc gcgagcgctt ctacaagctg caggaggact
     1321 gcaagaccat ctggcaggag agccgcaagg tgatgcgcac ccccgagagc cagctgttca
     1381 gcatcgagga catccaggag gtgcgcatgg gccaccgcac cgagggcctg gagaagttcg
     1441 cccgcgacgt gcccgaggac cgctgcttca gcatcgtgtt caaggaccag cgcaacaccc
     1501 tggacctgat cgcccccagc cccgccgacg cccagcactg ggtgctgggc ctgcacaaga
     1561 tcattcacca ttctggcagc atggatgggt acccagcttt cttgtacaaa gttggcatta
     1621 taagaaagca ttgcttatca atttgttgca acgaacaggt cactatcagt caaaataaaa
     1681 tcattatttg ccatccagct gatatcccct atagtgagtc gtattacatg gtcatagctg
     1741 tttcctggca gctctggccc gtgtctcaaa atctctgatg ttacattgca caagataaaa
     1801 taatatcatc atgaacaata aaactgtctg cttacataaa cagtaataca aggggtgtta
     1861 tgagccatat tcaacgggaa acgtcgaggc cgcgattaaa ttccaacatg gatgctgatt
     1921 tatatgggta taaatgggct cgcgataatg tcgggcaatc aggtgcgaca atctatcgct
     1981 tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca
     2041 atgatgttac agatgagatg gtcagactaa actggctgac ggaatttatg cctcttccga
     2101 ccatcaagca ttttatccgt actcctgatg atgcatggtt actcaccact gcgatccccg
     2161 gaaaaacagc attccaggta ttagaagaat atcctgattc aggtgaaaat attgttgatg
     2221 cgctggcagt gttcctgcgc cggttgcatt cgattcctgt ttgtaattgt ccttttaaca
     2281 gcgatcgcgt atttcgtctc gctcaggcgc aatcacgaat gaataacggt ttggttgatg
     2341 cgagtgattt tgatgacgag cgtaatggct ggcctgttga acaagtctgg aaagaaatgc
     2401 ataaactttt gccattctca ccggattcag tcgtcactca tggtgatttc tcacttgata
     2461 accttatttt tgacgagggg aaattaatag gttgtattga tgttggacga gtcggaatcg
     2521 cagaccgata ccaggatctt gccatcctat ggaactgcct cggtgagttt tctccttcat
     2581 tacagaaacg gctttttcaa aaatatggta ttgataatcc tgatatgaat aaattgcagt
     2641 ttcatttgat gctcgatgag tttttctaat cagaattggt taattggttg taacactggc
     2701 agagcattac gctgacttga cgggacggcg caagctcatg accaaaatcc cttaacgtga
     2761 gttacgcgtc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     2821 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     2881 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     2941 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     3001 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     3061 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     3121 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     3181 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     3241 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     3301 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     3361 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     3421 cctttttacg gttcctggcc ttttgctggc cttttgctca catgtt
//
