LOCUS       sequence-326618-        3536 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Three-fragment Gateway compatible flourophore for expression in C. 
            elegans.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3536)
  AUTHORS   Aljohani MD, El Mouridi S, Priyadarshini M, Vargas-Velazquez AM, 
            Frokjaer-Jensen C
  TITLE     Engineering rules that minimize germline silencing of transgenes in 
            simple extrachromosomal arrays in C. elegans.
  JOURNAL   Nat Commun. 2020 Dec 9;11(1):6300. doi: 10.1038/s41467-020-19898-0.
  PUBMED    33298957
REFERENCE   2  (bases 1 to 3536)
  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: "2020-12-9"; volume: "11(1):6300. doi"; pages: " 
            10.1038/s41467-020-19898-0"
COMMENT     Sequence Label: pCFJ2262A - [1-2] ENTR - Fluor - ce-mNeon (1 i...
FEATURES             Location/Qualifiers
     source          1..3536
                     /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      1060..1069
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(1661..1760)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1777..1796)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1778..1796)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1801..1817)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1801..1817)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1919..1938)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1930..2739
                     /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(1997..2016)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2886..3474
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3375..3394
                     /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 agcaggctcg gccggcagta ctagcggtgg cagtggaggt accggcggaa gcagttccaa
      721 ggtaacactt agtttttgtt gctctgttta aaaattaatt ttaagggaga ggaggacaac
      781 atggcctccc tcccagccac ccacgagctc cacatcttcg gctccatcaa cggagtcgac
      841 ttcgacatgg tcggacaagg aaccggaaac ccaaacgacg gatacgagga gctcaacctc
      901 aagtccacca aggtaagttt tcttgagacc tttactaact aactaggtct catagatttt
      961 cagggtgacc tccaattctc cccatggata ctcgtcccac acatcggata cggattccac
     1021 caatacctcc catacccaga cggaatgtcc ccattccaag ccgccatggt cgacgggtcc
     1081 ggataccaag tccaccgtac catgcaattc gaggacggag cctccctcac cgtcaactac
     1141 cgttacacct acgagggatc gcacatcaag gtaagttatg tagagacctt tactaactaa
     1201 ctaggtctca aacattttca gggagaggcc caagtcaagg gaaccggatt cccagccgac
     1261 ggaccagtca tgaccaactc cctcaccgcc gccgactggt gccgttccaa gaagacctac
     1321 ccaaacgaca agaccatcat ctccaccttc aagtggtcct acaccaccgg aaacggaaag
     1381 cgttaccgtt ccaccgcccg taccacctac accttcgcca agccaatggc cgccaactac
     1441 ctcaaggtaa gttctttaga gacctttact aactaactag gtctcagtaa ttttcagaac
     1501 caaccaatgt acgtcttccg taagaccgag ctcaagcact ccaagaccga gctcaacttc
     1561 aaggagtggc aaaaggcctt caccgacgtc atgggaatgg acgagctcta caagggtggc
     1621 acaggaggaa cgggcggtag tggaggcact ggtgccggca acccagcttt cttgtacaaa
     1681 gttggcatta taagaaagca ttgcttatca atttgttgca acgaacaggt cactatcagt
     1741 caaaataaaa tcattatttg ccatccagct gatatcccct atagtgagtc gtattacatg
     1801 gtcatagctg tttcctggca gctctggccc gtgtctcaaa atctctgatg ttacattgca
     1861 caagataaaa atatatcatc atgaacaata aaactgtctg cttacataaa cagtaataca
     1921 aggggtgtta tgagccatat tcaacgggaa acgtcgaggc cgcgattaaa ttccaacatg
     1981 gatgctgatt tatatgggta taaatgggct cgcgataatg tcgggcaatc aggtgcgaca
     2041 atctatcgct tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca tggcaaaggt
     2101 agcgttgcca atgatgttac agatgagatg gtcagactaa actggctgac ggaatttatg
     2161 cctcttccga ccatcaagca ttttatccgt actcctgatg atgcatggtt actcaccact
     2221 gcgatccccg gaaaaacagc attccaggta ttagaagaat atcctgattc aggtgaaaat
     2281 attgttgatg cgctggcagt gttcctgcgc cggttgcatt cgattcctgt ttgtaattgt
     2341 ccttttaaca gcgatcgcgt atttcgcctc gctcaggcgc aatcacgaat gaataacggt
     2401 ttggttgatg cgagtgattt tgatgacgag cgtaatggct ggcctgttga acaagtctgg
     2461 aaagaaatgc ataaactttt gccattctca ccggattcag tcgtcactca tggtgatttc
     2521 tcacttgata accttatttt tgacgagggg aaattaatag gttgtattga tgttggacga
     2581 gtcggaatcg cagaccgata ccaggatctt gccatcctat ggaactgcct cggtgagttt
     2641 tctccttcat tacagaaacg gctttttcaa aaatatggta ttgataatcc tgatatgaat
     2701 aaattgcagt ttcatttgat gctcgatgag tttttctaat cagaattggt taattggttg
     2761 taacactggc agagcattac gctgacttga cgggacggcg caagctcatg accaaaatcc
     2821 cttaacgtga gttacgcgtc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     2881 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     2941 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     3001 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     3061 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     3121 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     3181 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     3241 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     3301 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     3361 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     3421 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     3481 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgtt
//
