LOCUS       sequence-318854-        6194 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 6194)
  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 6194)
  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: pCFJ2441 - [1-2] ENTR - Fluor -
            ce-Dendra2(PAT...
FEATURES             Location/Qualifiers
     source          1..6194
                     /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"
     protein_bind    complement(4319..4418)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(4435..4454)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4436..4454)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4459..4475)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4459..4475)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4577..4596)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4588..5397
                     /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(4655..4674)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      5544..6132
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6033..6052
                     /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 gcagtaacac
      721 cccaggtaac acttagtttt tgttgctctg tttaaaaatt aattttaagg aatcaacctc
      781 atcaaggagg acatgcgtgt caaggtccac atggagggaa acgtcaacgg acacgccttc
      841 gtcatcgagg gagagggaaa gggaaagcca tacgagggaa cccaaaccgc caacctcacc
      901 gtcaaggtaa gttttctgtt ttatttactt gttttgagaa ttgggatttt tagctgaaaa
      961 attggatttt aagatgaaaa attgaaattt tcaactaaaa atttcaattt ttagctgaaa
     1021 agttaaattt ttagctaaag agtgggattt ttagttgaaa aatcgagatt tctagctaaa
     1081 aagtatgatt tttagctgaa agattggaat tttaagcaga aaaaattgta tttttagctg
     1141 ataaatcaaa ttttaagcgg aaaaatttga atttttagct aaaaaattcc attttcaact
     1201 aaaaatcgtg atttttagct ttaaaaattg gatttttagc tgaaaaatcg gatttttagc
     1261 tgaaaattga aattttttag ttaaaaagtg agatttttta acagaaaagt gagattttta
     1321 gttaaaaaat aatattttta gctaaaaaat acaattttca actaaagatc gtgattttta
     1381 gctttaaaaa ttggattttt agctaaaaaa tcggattttt agctgaaaat tgaaattttt
     1441 tagttaaaaa gtgggatttt aagatgaaaa gtgagatttt ttagctgaaa agtgagattt
     1501 tcagctaaaa aatacaattt tcaactgaaa aattgggatt tttagcttta aaaattggat
     1561 tttcagcttt aaaaattgga tttttagctg aaaagtgaga tttttagttg aaaaattgaa
     1621 atgtttagct gaaaaattgg aatttttaag taaaaaagtg ggattttaag atgaaaagtg
     1681 agatgttttt agctgaaaag tgagattttt agctaaaaaa tacaattttc aactgaaaaa
     1741 ttgggatttt tagctcaaaa gtgggacgaa agacgaaaaa aaaaatcgat tttttaccaa
     1801 aaattagatt ttcaggaggg agccccactc ccattctcct acgacatcct caccaccgcc
     1861 gtccactacg gaaaccgtgt cttcaccaag tacccagagg acatcccaga ctacttcaag
     1921 caatccttcc cagagggata ctcctgggag cgtaccatga ccttcgagga caagggaatc
     1981 tgcaccatcc gttccgacat ctccctcgag ggagactgct tcttccaaaa cgtccgtttc
     2041 aaggtaagtt atgtacgcga tttctcggcg aaaaaatgga cttttttgct caaaaactga
     2101 atttttcagc tttttttgca ctgaaaatgc tgaaaattgg tttttttgct gaaaattcac
     2161 cgaaaaaagc tgaaaatatt gcagaaatcg acgaaaaatg ccttttttgc tgaaatttca
     2221 gccagaaaat tgttcagaat cgcaatttcc tgccaaaatc gacgtttttc ggcgaaaatt
     2281 ggtagatttt gaaccgaaaa ttgtgaaaaa tggacttttt tgctcaaaaa cagacttttt
     2341 gagctttttt cacactgaaa atggtggaaa attggatttt ccagcagaaa attcggaaat
     2401 tttcaatttt tggcggaaat ttgacgaaaa atatatgaaa aacaccgatt tttgtctgaa
     2461 aaagttcaaa agccgaaaaa ttggccaaaa atctccaaaa tcggactttt ctggcgtttt
     2521 tcgtggaaaa attgaaattt ttttgaaaaa ttgagcaaaa atcgccgaaa aaccgctaaa
     2581 aaaactgccg aaattggcaa ttttgagctt aaaaaaactt gaaaaatgct caaaattacc
     2641 caattttcac cggaaattga gctaaaatcg tcgaaaaaga ctgaattttc aatttttagc
     2701 tgaaaatctg cctattttcg gctaaaattg ctttaaaaac attaaaattc aaattttttg
     2761 gctaaaaata ctcagattgc cctttttttt ctccaaaaat gacaaaaaaa aaatcaatca
     2821 gaaaaatgcg atttttttta caaaaattgg cttcaaaacg tctaaattct ccctaatttt
     2881 tgctgctgaa aattgtgcaa aaatcaccga aaaatgacga ttttaggctg aaaatcttca
     2941 aaaaaagctc aaaattcccc acttttcacc ggaaaaagac gaaaaaagct ccaaatttat
     3001 cgattaacat tttcagggaa ccaacttccc accaaacgga ccagtcatgc aaaagaagac
     3061 cctcaagtgg gagccatcca ccgagaagct ccacgtccgt gacggactcc tcgtcggaaa
     3121 catcaacatg gccctcctcc tcgagggagg aggacactac ctctgcgact tcaaggtaag
     3181 ttcttttttt tgctcgtttt tgagtgaaaa attgatggaa ttcacaaaaa atggccaaag
     3241 aatcctcaaa attggccgaa aattctgaaa aatgacctaa aaatcaccga aaaacggcca
     3301 aaaattatct gaaaatcgcc gaaaatctca aattttgagg aaattcccgc taaaaattaa
     3361 taaaaatcac cgaaaaaagg ccaaaaatta cctaataaag gccgaaaatc gcctaaaaag
     3421 gccaaaaatt ctgaaaaatg gccaaatttc aatttttcac acacaaaaaa tccctgaatt
     3481 ttctattttt actcgaaaaa ctgtggattt acgggggtaa aaattcagtt tttagtgcta
     3541 tttcacaaga aaaacacagc attttcgaca aaaactccga tttttaagcc aaaaatttga
     3601 ggtttttagt tttttacgtg aaaacctgaa ctttttgcca aaaaaacact tttttttaga
     3661 ctaaaaatca attttcgatc aaaaatcact aattttgcct gaaaaaatat agatttttcc
     3721 actttttcac tgaaaaaaaa ctggattttc actagaaaat caagttgttt ttagaccaaa
     3781 aattttattt ttccgctgaa aattatcgtt tttcagtcaa ttttcagcta aaaatccctg
     3841 aaattctgga ttttcagcca aaaaactcga aattttccaa aaaaaccccg tttaaaaatg
     3901 agtttagact gaaaattccg atttttgagc caaaaatttg atttttaatc gaaaaaaatt
     3961 ggattttcaa ccaaaaaatc agtttttcag tgatttttag agtaaaaatc actgatttta
     4021 acacaaaatt ccagtttttc gtgatttttc actagaaaat cccgattttt ccccaaaaaa
     4081 tccatttttc cgactaaaaa tccccaaagt aattttcaga ccacctacaa ggccaagaag
     4141 gtcgtccaac tcccagacgc ccacttcgtc gaccaccgta tcgagatcct cggaaacgac
     4201 tccgactaca acaaggtcaa gctctacgag cacgccgtcg cccgttactc cccactccca
     4261 tcccaagtct ggggtggcac aggaggaacg ggcggtagtg gaggcactgg tgccggcaac
     4321 ccagctttct tgtacaaagt tggcattata agaaagcatt gcttatcaat ttgttgcaac
     4381 gaacaggtca ctatcagtca aaataaaatc attatttgcc atccagctga tatcccctat
     4441 agtgagtcgt attacatggt catagctgtt tcctggcagc tctggcccgt gtctcaaaat
     4501 ctctgatgtt acattgcaca agataaaaat atatcatcat gaacaataaa actgtctgct
     4561 tacataaaca gtaatacaag gggtgttatg agccatattc aacgggaaac gtcgaggccg
     4621 cgattaaatt ccaacatgga tgctgattta tatgggtata aatgggctcg cgataatgtc
     4681 gggcaatcag gtgcgacaat ctatcgcttg tatgggaagc ccgatgcgcc agagttgttt
     4741 ctgaaacatg gcaaaggtag cgttgccaat gatgttacag atgagatggt cagactaaac
     4801 tggctgacgg aatttatgcc tcttccgacc atcaagcatt ttatccgtac tcctgatgat
     4861 gcatggttac tcaccactgc gatccccgga aaaacagcat tccaggtatt agaagaatat
     4921 cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg gttgcattcg
     4981 attcctgttt gtaattgtcc ttttaacagc gatcgcgtat ttcgcctcgc tcaggcgcaa
     5041 tcacgaatga ataacggttt ggttgatgcg agtgattttg atgacgagcg taatggctgg
     5101 cctgttgaac aagtctggaa agaaatgcat aaacttttgc cattctcacc ggattcagtc
     5161 gtcactcatg gtgatttctc acttgataac cttatttttg acgaggggaa attaataggt
     5221 tgtattgatg ttggacgagt cggaatcgca gaccgatacc aggatcttgc catcctatgg
     5281 aactgcctcg gtgagttttc tccttcatta cagaaacggc tttttcaaaa atatggtatt
     5341 gataatcctg atatgaataa attgcagttt catttgatgc tcgatgagtt tttctaatca
     5401 gaattggtta attggttgta acactggcag agcattacgc tgacttgacg ggacggcgca
     5461 agctcatgac caaaatccct taacgtgagt tacgcgtcgt tccactgagc gtcagacccc
     5521 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     5581 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     5641 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     5701 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     5761 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     5821 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     5881 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     5941 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     6001 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     6061 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     6121 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     6181 tttgctcaca tgtt
//
