LOCUS       sequence-318041-        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: pCFJ2438 - [1-2] ENTR - Fluor -
            ce-mCardinal((...
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 accgagggag agggaaagcc atacgaggga acccaaaccc aacgtatcaa
      361 ggtcgtcgag ggaggaccac tcccattcgc cttcgacatc ctcgccacct gcttcatgta
      421 cggatcgaag gtaagttttc tgttttattt acttgtttct gagaattggg atttttagct
      481 gaaaaattgg attttaagat gaaaaattga aattttcaac taaaaatttc aatttttagc
      541 tgaaaagtta aatttttagc taaagagtgg gatttttagt tgaaaaatcg agatttctag
      601 ctaaaaagta tgatttttag ctgaaagatt ggaattttaa gcagaaaaaa ttgtattttt
      661 agctgataaa tcaaatttta agcggaaaaa tttgaatttt tagctaaaaa attccatttt
      721 caactaaaaa tcgtgatttt tagctttaaa aattggattt ttagctgaaa aatcggattt
      781 ttagctgaaa attgaaattt tttagttaaa aagtgagatt ttttaacaga aaagtgagat
      841 ttttagttaa aaaataatat ttttagctaa aaaatacaat tttcaactaa agatcgtgat
      901 ttttagcttt aaaaattgga tttttagcta aaaaatcgga tttttagctg aaaattgaaa
      961 ttttttagtt aaaaagtggg attttaagat gaaaagtgag attttttagc tgaaaagtga
     1021 gattttcagc taaaaaatac aattttcaac tgaaaaattg ggatttttag ctttaaaaat
     1081 tggattttca gctttaaaaa ttggattttt agctgaaaag tgagattttt agttgaaaaa
     1141 ttgaaatgtt tagctgaaaa attggaattt ttaagtaaaa aagtgggatt ttaagatgaa
     1201 aagtgagatg tttttagctg aaaagtgaga tttttagcta aaaaatacaa ttttcaactg
     1261 aaaaattggg atttttagct caaaagtggg acgaaagacg aaaaaaaaaa tcgatttttt
     1321 accaaaaatt agattttcag accttcatca accacaccca aggaatccca gacttcttca
     1381 agcaatcctt cccagaggga ttcacctggg agcgtgtcac cacctacgag gacggaggag
     1441 tcctcaccgt cacccaagac acctccctcc aaggtaagtt atgtacgcga tttctcggcg
     1501 aaaaaatgga cttttttgct caaaaactga atttttcagc tttttttgca ctgaaaatgc
     1561 tgaaaattgg tttttttgct gaaaattcac cgaaaaaagc tgaaaatatt gcagaaatcg
     1621 acgaaaaatg ccttttttgc tgaaatttca gccagaaaat tgttcagaat cgcaatttcc
     1681 tgccaaaatc gacgtttttc ggcgaaaatt ggtagatttt gaaccgaaaa ttgtgaaaaa
     1741 tggacttttt tgctcaaaaa cagacttttt gagctttttt cacactgaaa atggtggaaa
     1801 attggatttt ccagcagaaa attcggaaat tttcaatttt tggcggaaat ttgacgaaaa
     1861 atatatgaaa aacaccgatt tttgtctgaa aaagttcaaa agccgaaaaa ttggccaaaa
     1921 atctccaaaa tcggactttt ctggcgtttt tcgtggaaaa attgaaattt ttttgaaaaa
     1981 ttgagcaaaa atcgccgaaa aaccgctaaa aaaactgccg aaattggcaa ttttgagctt
     2041 aaaaaaactt gaaaaatgct caaaattacc caattttcac cggaaattga gctaaaatcg
     2101 tcgaaaaaga ctgaattttc aatttttagc tgaaaatctg cctattttcg gctaaaattg
     2161 ctttaaaaac attaaaattc aaattttttg gctaaaaata ctcagattgc cctttttttt
     2221 ctccaaaaat gacaaaaaaa aaatcaatca gaaaaatgcg atttttttta caaaaattgg
     2281 cttcaaaacg tctaaattct ccctaatttt tgctgctgaa aattgtgcaa aaatcaccga
     2341 aaaatgacga ttttaggctg aaaatcttca aaaaaagctc aaaattcccc acttttcacc
     2401 ggaaaaagac gaaaaaagct ccaaatttat cgattaacat tttcagacgg atgcctcatc
     2461 tacaacgtca agctccgtgg agtcaacttc ccatccaacg gaccagtcat gcaaaagaag
     2521 accctcggat gggaggccac caccgagacg ctctacccag ccgacggagg actcgaggga
     2581 cgttgcgaca tggccctcaa gctcgtcgga ggaggacacc tccactgcaa cctcaaggta
     2641 agttcttttt tttgctcgtt tttgagtgaa aaattgatgg aattcacaaa aaatggccaa
     2701 agaatcctca aaattggccg aaaattctga aaaatgacct aaaaatcacc gaaaaacggc
     2761 caaaaattat ctgaaaatcg ccgaaaatct caaattttga ggaaattccc gctaaaaatt
     2821 aataaaaatc accgaaaaaa ggccaaaaat tacctaataa aggccgaaaa tcgcctaaaa
     2881 aggccaaaaa ttctgaaaaa tggccaaatt tcaatttttc acacacaaaa aatccctgaa
     2941 ttttctattt ttactcgaaa aactgtggat ttacgggggt aaaaattcag tttttagtgc
     3001 tatttcacaa gaaaaacaca gcattttcga caaaaactcc gatttttaag ccaaaaattt
     3061 gaggttttta gttttttacg tgaaaacctg aactttttgc caaaaaaaca ctttttttta
     3121 gactaaaaat caattttcga tcaaaaatca ctaattttgc ctgaaaaaat atagattttt
     3181 ccactttttc actgaaaaaa aactggattt tcactagaaa atcaagttgt ttttagacca
     3241 aaaattttat ttttccgctg aaaattatcg tttttcagtc aattttcagc taaaaatccc
     3301 tgaaattctg gattttcagc caaaaaactc gaaattttcc aaaaaaaccc cgtttaaaaa
     3361 tgagtttaga ctgaaaattc cgatttttga gccaaaaatt tgatttttaa tcgaaaaaaa
     3421 ttggattttc aaccaaaaaa tcagtttttc agtgattttt agagtaaaaa tcactgattt
     3481 taacacaaaa ttccagtttt tcgtgatttt tcactagaaa atcccgattt ttccccaaaa
     3541 aatccatttt tccgactaaa aatccccaaa gtaattttca gaccacctac cgttccaaga
     3601 agccagccaa gaacctcaag atgccaggag tctacttcgt cgaccgtcgt ctcgagcgta
     3661 tcaaggaggc cgacaacgag 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
//
