LOCUS       sequence-318022-        6234 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 6234)
  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 6234)
  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: pCFJ2437 - [1-2] ENTR - Fluor -
            ce-tagRFP-T(PA...
FEATURES             Location/Qualifiers
     source          1..6234
                     /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"
     protein_bind    complement(3823..3922)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3939..3958)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3940..3958)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3963..3979)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3963..3979)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4081..4100)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4092..4901
                     /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(4159..4178)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      5048..5636
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5537..5556
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5790..5807
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(5966..5993)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(6085..6171)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     join(6220..6234,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(6234,1..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 agtactagcg gtggcagtgg aggtaccggc ggaagcagtt
      181 ccaaggtaac acttagtttt tgttgctctg tttaaaaatt aattttaagg gagaggagct
      241 catcaaggag aacatgcaca tgaagctcta catggaggga accgtcaaca accaccactt
      301 caagtgcacc tccgagggag agggaaagcc atacgaggga acccaaacca tgcgtatcaa
      361 ggtcgtcgag ggaggaccac tcccattcgc cttcgacatc ctcgccacct ccttcatgta
      421 cggatcgcgt accttcatca accacaccca aggtaagttt tctgttttat ttacttgttt
      481 ctgagaattg ggatttttag ctgaaaaatt ggattttaag atgaaaaatt gaaattttca
      541 actaaaaatt tcaattttta gctgaaaagt taaattttta gctaaagagt gggattttta
      601 gttgaaaaat cgagatttct agctaaaaag tatgattttt agctgaaaga ttggaatttt
      661 aagcagaaaa aattgtattt ttagctgata aatcaaattt taagcggaaa aatttgaatt
      721 tttagctaaa aaattccatt ttcaactaaa aatcgtgatt tttagcttta aaaattggat
      781 ttttagctga aaaatcggat ttttagctga aaattgaaat tttttagtta aaaagtgaga
      841 ttttttaaca gaaaagtgag atttttagtt aaaaaataat atttttagct aaaaaataca
      901 attttcaact aaagatcgtg atttttagct ttaaaaattg gatttttagc taaaaaatcg
      961 gatttttagc tgaaaattga aattttttag ttaaaaagtg ggattttaag atgaaaagtg
     1021 agatttttta gctgaaaagt gagattttca gctaaaaaat acaattttca actgaaaaat
     1081 tgggattttt agctttaaaa attggatttt cagctttaaa aattggattt ttagctgaaa
     1141 agtgagattt ttagttgaaa aattgaaatg tttagctgaa aaattggaat ttttaagtaa
     1201 aaaagtggga ttttaagatg aaaagtgaga tgtttttagc tgaaaagtga gatttttagc
     1261 taaaaaatac aattttcaac tgaaaaattg ggatttttag ctcaaaagtg ggacgaaaga
     1321 cgaaaaaaaa aatcgatttt ttaccaaaaa ttagattttc aggaatccca gacttcttca
     1381 agcaatcctt cccagaggga ttcacctggg agcgtgtcac cacctacgag gacggaggag
     1441 tcctcaccgc cacccaagac acctccctcc aagacggatg cctcatctac aacgtcaagg
     1501 taagttatgt acgcgatttc tcggcgaaaa aatggacttt tttgctcaaa aactgaattt
     1561 ttcagctttt tttgcactga aaatgctgaa aattggtttt tttgctgaaa attcaccgaa
     1621 aaaagctgaa aatattgcag aaatcgacga aaaatgcctt ttttgctgaa atttcagcca
     1681 gaaaattgtt cagaatcgca atttcctgcc aaaatcgacg tttttcggcg aaaattggta
     1741 gattttgaac cgaaaattgt gaaaaatgga cttttttgct caaaaacaga ctttttgagc
     1801 ttttttcaca ctgaaaatgg tggaaaattg gattttccag cagaaaattc ggaaattttc
     1861 aatttttggc ggaaatttga cgaaaaatat atgaaaaaca ccgatttttg tctgaaaaag
     1921 ttcaaaagcc gaaaaattgg ccaaaaatct ccaaaatcgg acttttctgg cgtttttcgt
     1981 ggaaaaattg aaattttttt gaaaaattga gcaaaaatcg ccgaaaaacc gctaaaaaaa
     2041 ctgccgaaat tggcaatttt gagcttaaaa aaacttgaaa aatgctcaaa attacccaat
     2101 tttcaccgga aattgagcta aaatcgtcga aaaagactga attttcaatt tttagctgaa
     2161 aatctgccta ttttcggcta aaattgcttt aaaaacatta aaattcaaat tttttggcta
     2221 aaaatactca gattgccctt ttttttctcc aaaaatgaca aaaaaaaaat caatcagaaa
     2281 aatgcgattt tttttacaaa aattggcttc aaaacgtcta aattctccct aatttttgct
     2341 gctgaaaatt gtgcaaaaat caccgaaaaa tgacgatttt aggctgaaaa tcttcaaaaa
     2401 aagctcaaaa ttccccactt ttcaccggaa aaagacgaaa aaagctccaa atttatcgat
     2461 taacattttc agatccgtgg agtcaacttc ccatccaacg gaccagtcat gcaaaagaag
     2521 accctcggat gggaggccaa caccgagatg ctctacccag ccgacggagg actggaggga
     2581 cgtaccgaca tggccctcaa gctcgtcgga ggaggacacc tcatctgcaa cttcaagacc
     2641 acctaccgtt ccaagaagcc agccaaggta agttcttttt tttgctcgtt tttgagtgaa
     2701 aaattgatgg aattcacaaa aaatggccaa agaatcctca aaattggccg aaaattctga
     2761 aaaatgacct aaaaatcacc gaaaaacggc caaaaattat ctgaaaatcg ccgaaaatct
     2821 caaattttga ggaaattccc gctaaaaatt aataaaaatc accgaaaaaa ggccaaaaat
     2881 tacctaataa aggccgaaaa tcgcctaaaa aggccaaaaa ttctgaaaaa tggccaaatt
     2941 tcaatttttc acacacaaaa aatccctgaa ttttctattt ttactcgaaa aactgtggat
     3001 ttacgggggt aaaaattcag tttttagtgc tatttcacaa gaaaaacaca gcattttcga
     3061 caaaaactcc gatttttaag ccaaaaattt gaggttttta gttttttacg tgaaaacctg
     3121 aactttttgc caaaaaaaca ctttttttta gactaaaaat caattttcga tcaaaaatca
     3181 ctaattttgc ctgaaaaaat atagattttt ccactttttc actgaaaaaa aactggattt
     3241 tcactagaaa atcaagttgt ttttagacca aaaattttat ttttccgctg aaaattatcg
     3301 tttttcagtc aattttcagc taaaaatccc tgaaattctg gattttcagc caaaaaactc
     3361 gaaattttcc aaaaaaaccc cgtttaaaaa tgagtttaga ctgaaaattc cgatttttga
     3421 gccaaaaatt tgatttttaa tcgaaaaaaa ttggattttc aaccaaaaaa tcagtttttc
     3481 agtgattttt agagtaaaaa tcactgattt taacacaaaa ttccagtttt tcgtgatttt
     3541 tcactagaaa atcccgattt ttccccaaaa aatccatttt tccgactaaa aatccccaaa
     3601 gtaattttca gaacctcaag atgccaggag tctactacgt cgaccaccgt ctggagcgta
     3661 tcaaggaggc cgacaaggag acgtacgtcg agcaacacga ggtcgccgtc gcccgttact
     3721 gcgacctccc atccaagctc ggacacaagc tcaacggaat ggacgagctc tacaagggtg
     3781 gcacaggagg aacgggcggt agtggaggca ctggtgccgg caacccagct ttcttgtaca
     3841 aagttggcat tataagaaag cattgcttat caatttgttg caacgaacag gtcactatca
     3901 gtcaaaataa aatcattatt tgccatccag ctgatatccc ctatagtgag tcgtattaca
     3961 tggtcatagc tgtttcctgg cagctctggc ccgtgtctca aaatctctga tgttacattg
     4021 cacaagataa aaatatatca tcatgaacaa taaaactgtc tgcttacata aacagtaata
     4081 caaggggtgt tatgagccat attcaacggg aaacgtcgag gccgcgatta aattccaaca
     4141 tggatgctga tttatatggg tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga
     4201 caatctatcg cttgtatggg aagcccgatg cgccagagtt gtttctgaaa catggcaaag
     4261 gtagcgttgc caatgatgtt acagatgaga tggtcagact aaactggctg acggaattta
     4321 tgcctcttcc gaccatcaag cattttatcc gtactcctga tgatgcatgg ttactcacca
     4381 ctgcgatccc cggaaaaaca gcattccagg tattagaaga atatcctgat tcaggtgaaa
     4441 atattgttga tgcgctggca gtgttcctgc gccggttgca ttcgattcct gtttgtaatt
     4501 gtccttttaa cagcgatcgc gtatttcgcc tcgctcaggc gcaatcacga atgaataacg
     4561 gtttggttga tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct
     4621 ggaaagaaat gcataaactt ttgccattct caccggattc agtcgtcact catggtgatt
     4681 tctcacttga taaccttatt tttgacgagg ggaaattaat aggttgtatt gatgttggac
     4741 gagtcggaat cgcagaccga taccaggatc ttgccatcct atggaactgc ctcggtgagt
     4801 tttctccttc attacagaaa cggctttttc aaaaatatgg tattgataat cctgatatga
     4861 ataaattgca gtttcatttg atgctcgatg agtttttcta atcagaattg gttaattggt
     4921 tgtaacactg gcagagcatt acgctgactt gacgggacgg cgcaagctca tgaccaaaat
     4981 cccttaacgt gagttacgcg tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     5041 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     5101 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     5161 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     5221 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     5281 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     5341 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     5401 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     5461 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     5521 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     5581 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     5641 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     5701 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     5761 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
     5821 gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg
     5881 acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatacg cgtaccgcta
     5941 gccaggaaga gtttgtagaa acgcaaaaag gccatccgtc aggatggcct tctgcttagt
     6001 ttgatgcctg gcagtttatg gcgggcgtcc tgcccgccac cctccgggcc gttgcttcac
     6061 aacgttcaaa tccgctcccg gcggatttgt cctactcagg agagcgttca ccgacaaaca
     6121 acagataaaa cgaaaggccc agtcttccga ctgagccttt cgttttattt gatgcctggc
     6181 agttccctac tctcgcgtta acgctagcat ggatgttttc ccagtcacga cgtt
//
