LOCUS       sequence-318016-        4547 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 4547)
  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 4547)
  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: pCFJ2092 - [1-2] ENTR - Fluor - mNeon(PATC)
FEATURES             Location/Qualifiers
     source          1..4547
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    33..132
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     regulatory      778..787
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(2136..2235)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2252..2271)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2253..2271)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2276..2292)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2276..2292)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2394..2413)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2405..3214
                     /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(2472..2491)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3361..3949
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3850..3869
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4103..4120
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(4279..4306)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4398..4484)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     join(4533..4547,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(4547,1..17)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
ORIGIN
        1 gtaaaacgac ggccagtctt aagctcgggc cccaaataat gattttattt tgactgatag
       61 tgacctgttc gttgcaacaa attgatgagc aatgcttttt tataatgcca actttgtaca
      121 aaaaagcagg ctcggccggc ggatcatcct ccacatcatc ctccacatca tcctcagtct
      181 ccaaggtaac acttagtttt tgttgctctg tttaaaaatt aattttaagg gagaggagga
      241 caacatggcc tccctcccag ccacccacga gctccacatc ttcggctcca tcaacggagt
      301 cgacttcgac atggtcggac aaggaaccgg aaacccaaac gacggatacg aggagctcaa
      361 cctcaagtcc accaaggtaa gttttcttat gggaaagaag gaaaaaaccg agattttact
      421 tgaaaaattg aatttttcgc gggattttca ccaaaaattg ttgaatattc attatttcac
      481 gctgtaaaac aaaaaaaaaa aaaatcaaaa actacgttga aatcgcgttt ttaagcgaat
      541 tttcttcaga attgccagat tttaacccca aattttgcag tttttaaata aaatttcacc
      601 ttttcggctc aaattgtaga ttttcctgaa aatttagtac aaaaaacaat ttcctcgtaa
      661 atttttcaaa tagattttca gggtgacctc caattctccc catggatact cgtcccacac
      721 atcggatacg gattccacca atacctccca tacccagacg gaatgtcccc attccaagcc
      781 gccatggtcg acgggtccgg ataccaagtc caccgtacca tgcaattcga ggacggagcc
      841 tccctcaccg tcaactaccg ttacacctac gagggatcgc acatcaaggt aagttatgtc
      901 tacctgcctg cctaccgcct aaattttttg tgaagtttct tcaaaaaatc cagaaaaaaa
      961 aacaattttc atacgatttt ttcccttaaa attgtgaatt ttcatgcttt ttagccccaa
     1021 aagtcattat ttgagaaaaa atttcataca aaaaagtttt gagaaataca caatttttta
     1081 aatgtaattt tcaaattttc aattttcaac tagaaaattc acaaaacttg taaattttgg
     1141 accaaaacat ttatacaatt actttttttg aatctaataa ctacaataac tacaaattta
     1201 acattttcag ggagaggccc aagtcaaggg aaccggattc ccagccgacg gaccagtcat
     1261 gaccaactcc ctcaccgccg ccgactggtg ccgttccaag aagacctacc caaacgacaa
     1321 gaccatcatc tccaccttca agtggtccta caccaccgga aacggaaagc gttaccgttc
     1381 caccgcccgt accacctaca ccttcgccaa gccaatggcc gccaactacc tcaaggtaag
     1441 ttcttttttt gaaaagtgca gtttgtagtc taattttcat ttattttctt tttaaaaaac
     1501 gcatcaattt ttaaatattt tgggacaaaa tccgaaaact gtactaattt gtagtttgta
     1561 aaattaaaaa aaaaacgcaa aaaaatgttt tcaaaaatgc ttagaataaa aataaaattt
     1621 ctaaaaaatt gaacaaaata aacatttaaa aattcaaaaa gtttgaaaaa atgcaaagtt
     1681 ttttaatgca aaaaatattt aatgtttaaa caaatttaaa aaatgttgta agaatatgtt
     1741 tagagacagt ttttaaatgt taaattgcaa aaaaaaaaag caaaaaaaaa tttgaaaaat
     1801 gtaacagaaa attttaatta aaaaagtcgt gtttttttca aacctgcaat tctttgtaat
     1861 tttatttata aagagaaaca aatcaaattg tatattacga aaaacgatac taaaattcga
     1921 aaatttgcgg ttttttgcgt aaaaaatacg gttccgtaat tttcagaacc aaccaatgta
     1981 cgtcttccgt aagaccgagc tcaagcactc caagaccgag ctcaacttca aggagtggca
     2041 aaaggccttc accgacgtca tgggaatgga cgagctctac aagtcgggaa cttctacgac
     2101 ttcggtaacg tcatccggta cttccccagc cggcaaccca gctttcttgt acaaagttgg
     2161 cattataaga aagcattgct tatcaatttg ttgcaacgaa caggtcacta tcagtcaaaa
     2221 taaaatcatt atttgccatc cagctgatat cccctatagt gagtcgtatt acatggtcat
     2281 agctgtttcc tggcagctct ggcccgtgtc tcaaaatctc tgatgttaca ttgcacaaga
     2341 taaaaatata tcatcatgaa caataaaact gtctgcttac ataaacagta atacaagggg
     2401 tgttatgagc catattcaac gggaaacgtc gaggccgcga ttaaattcca acatggatgc
     2461 tgatttatat gggtataaat gggctcgcga taatgtcggg caatcaggtg cgacaatcta
     2521 tcgcttgtat gggaagcccg atgcgccaga gttgtttctg aaacatggca aaggtagcgt
     2581 tgccaatgat gttacagatg agatggtcag actaaactgg ctgacggaat ttatgcctct
     2641 tccgaccatc aagcatttta tccgtactcc tgatgatgca tggttactca ccactgcgat
     2701 ccccggaaaa acagcattcc aggtattaga agaatatcct gattcaggtg aaaatattgt
     2761 tgatgcgctg gcagtgttcc tgcgccggtt gcattcgatt cctgtttgta attgtccttt
     2821 taacagcgat cgcgtatttc gcctcgctca ggcgcaatca cgaatgaata acggtttggt
     2881 tgatgcgagt gattttgatg acgagcgtaa tggctggcct gttgaacaag tctggaaaga
     2941 aatgcataaa cttttgccat tctcaccgga ttcagtcgtc actcatggtg atttctcact
     3001 tgataacctt atttttgacg aggggaaatt aataggttgt attgatgttg gacgagtcgg
     3061 aatcgcagac cgataccagg atcttgccat cctatggaac tgcctcggtg agttttctcc
     3121 ttcattacag aaacggcttt ttcaaaaata tggtattgat aatcctgata tgaataaatt
     3181 gcagtttcat ttgatgctcg atgagttttt ctaatcagaa ttggttaatt ggttgtaaca
     3241 ctggcagagc attacgctga cttgacggga cggcgcaagc tcatgaccaa aatcccttaa
     3301 cgtgagttac gcgtcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     3361 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     3421 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     3481 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     3541 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     3601 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     3661 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     3721 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     3781 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     3841 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     3901 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     3961 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     4021 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
     4081 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
     4141 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
     4201 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat acgcgtaccg ctagccagga
     4261 agagtttgta gaaacgcaaa aaggccatcc gtcaggatgg ccttctgctt agtttgatgc
     4321 ctggcagttt atggcgggcg tcctgcccgc caccctccgg gccgttgctt cacaacgttc
     4381 aaatccgctc ccggcggatt tgtcctactc aggagagcgt tcaccgacaa acaacagata
     4441 aaacgaaagg cccagtcttc cgactgagcc tttcgtttta tttgatgcct ggcagttccc
     4501 tactctcgcg ttaacgctag catggatgtt ttcccagtca cgacgtt
//
