LOCUS       sequence-317997-        6197 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 6197)
  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 6197)
  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: pCFJ2451 - [1-2] ENTR - Fluor -
            ce-tagBFP2(PAT...
FEATURES             Location/Qualifiers
     source          1..6197
                     /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(3786..3885)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3902..3921)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3903..3921)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3926..3942)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3926..3942)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4044..4063)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4055..4864
                     /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(4122..4141)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      5011..5599
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5500..5519
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5753..5770
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(5929..5956)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(6048..6134)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     join(6183..6197,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(6197,1..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 ccgagctcat caaggtaaca cttagttttt gttgctctgt ttaaaaatta attttaagga
      241 gaacatgcac atgaagctct acatggaggg aaccgtcgac aaccaccact tcaagtgcac
      301 ctccgaggga gagggaaagc catacgaggg aacccaaacc atgcgtatca aggtcgtcga
      361 gggaggacca ctcccattcg ccttcgacat cctcgccacc tccttcctct acggatcgaa
      421 gaccttcatc aaccacaccc aaggaatccc agacttcttc aagcaatcct tcccaggtaa
      481 gttttctgtt ttatttactt gttttgagaa ttgggatttt tagctgaaaa attggatttt
      541 aagatgaaaa attgaaattt tcaactaaaa atttcaattt ttagctgaaa agttaaattt
      601 ttagctaaag agtgggattt ttagttgaaa aatcgagatt tctagctaaa aagtatgatt
      661 tttagctgaa agattggaat tttaagcaga aaaaattgta tttttagctg ataaatcaaa
      721 ttttaagcgg aaaaatttga atttttagct aaaaaattcc attttcaact aaaaatcgtg
      781 atttttagct ttaaaaattg gatttttagc tgaaaaatcg gatttttagc tgaaaattga
      841 aattttttag ttaaaaagtg agatttttta acagaaaagt gagattttta gttaaaaaat
      901 aatattttta gctaaaaaat acaattttca actaaagatc gtgattttta gctttaaaaa
      961 ttggattttt agctaaaaaa tcggattttt agctgaaaat tgaaattttt tagttaaaaa
     1021 gtgggatttt aagatgaaaa gtgagatttt ttagctgaaa agtgagattt tcagctaaaa
     1081 aatacaattt tcaactgaaa aattgggatt tttagcttta aaaattggat tttcagcttt
     1141 aaaaattgga tttttagctg aaaagtgaga tttttagttg aaaaattgaa atgtttagct
     1201 gaaaaattgg aatttttaag taaaaaagtg ggattttaag atgaaaagtg agatgttttt
     1261 agctgaaaag tgagattttt agctaaaaaa tacaattttc aactgaaaaa ttgggatttt
     1321 tagctcaaaa gtgggacgaa agacgaaaaa aaaaatcgat tttttaccaa aaattagatt
     1381 ttcagaggga ttcacctggg agcgtgtcac cacctacgag gacggaggag tcctcaccgc
     1441 cacccaagac acctccctcc aagacggatg cctcatctac aacgtcaaga tccgtggagt
     1501 caacttcacc tccaacggac cagtcatgca aaaggtaagt tatgtacgcg atttctcggc
     1561 gaaaaaatgg acttttttgc tcaaaaactg aatttttcag ctttttttgc actgaaaatg
     1621 ctgaaaattg gtttttttgc tgaaaattca ccgaaaaaag ctgaaaatat tgcagaaatc
     1681 gacgaaaaat gccttttttg ctgaaatttc agccagaaaa ttgttcagaa tcgcaatttc
     1741 ctgccaaaat cgacgttttt cggcgaaaat tggtagattt tgaaccgaaa attgtgaaaa
     1801 atggactttt ttgctcaaaa acagactttt tgagcttttt tcacactgaa aatggtggaa
     1861 aattggattt tccagcagaa aattcggaaa ttttcaattt ttggcggaaa tttgacgaaa
     1921 aatatatgaa aaacaccgat ttttgtctga aaaagttcaa aagccgaaaa attggccaaa
     1981 aatctccaaa atcggacttt tctggcgttt ttcgtggaaa aattgaaatt tttttgaaaa
     2041 attgagcaaa aatcgccgaa aaaccgctaa aaaaactgcc gaaattggca attttgagct
     2101 taaaaaaact tgaaaaatgc tcaaaattac ccaattttca ccggaaattg agctaaaatc
     2161 gtcgaaaaag actgaatttt caatttttag ctgaaaatct gcctattttc ggctaaaatt
     2221 gctttaaaaa cattaaaatt caaatttttt ggctaaaaat actcagattg cccttttttt
     2281 tctccaaaaa tgacaaaaaa aaaatcaatc agaaaaatgc gatttttttt acaaaaattg
     2341 gcttcaaaac gtctaaattc tccctaattt ttgctgctga aaattgtgca aaaatcaccg
     2401 aaaaatgacg attttaggct gaaaatcttc aaaaaaagct caaaattccc cacttttcac
     2461 cggaaaaaga cgaaaaaagc tccaaattta tcgattaaca ttttcagaag accctcggat
     2521 gggaggcctt caccgagacg ctctacccag ccgacggagg actggaggga cgtaacgaca
     2581 tggccctcaa gctcgtcgga ggatcgcacc tcatcgccaa cgccaagacc acctaccgtt
     2641 ccaagaagcc agccaagaac ctcaagatgc cagtctacta cgtcgactac cgtctggagc
     2701 gtaaggtaag ttcttttttt tgctcgtttt tgagtgaaaa attgatggaa ttcacaaaaa
     2761 atggccaaag aatcctcaaa attggccgaa aattctgaaa aatgacctaa aaatcaccga
     2821 aaaacggcca aaaattatct gaaaatcgcc gaaaatctca aattttgagg aaattcccgc
     2881 taaaaattaa taaaaatcac cgaaaaaagg ccaaaaatta cctaataaag gccgaaaatc
     2941 gcctaaaaag gccaaaaatt ctgaaaaatg gccaaatttc aatttttcac acacaaaaaa
     3001 tccctgaatt ttctattttt actcgaaaaa ctgtggattt acgggggtaa aaattcagtt
     3061 tttagtgcta tttcacaaga aaaacacagc attttcgaca aaaactccga tttttaagcc
     3121 aaaaatttga ggtttttagt tttttacgtg aaaacctgaa ctttttgcca aaaaaacact
     3181 tttttttaga ctaaaaatca attttcgatc aaaaatcact aattttgcct gaaaaaatat
     3241 agatttttcc actttttcac tgaaaaaaaa ctggattttc actagaaaat caagttgttt
     3301 ttagaccaaa aattttattt ttccgctgaa aattatcgtt tttcagtcaa ttttcagcta
     3361 aaaatccctg aaattctgga ttttcagcca aaaaactcga aattttccaa aaaaaccccg
     3421 tttaaaaatg agtttagact gaaaattccg atttttgagc caaaaatttg atttttaatc
     3481 gaaaaaaatt ggattttcaa ccaaaaaatc agtttttcag tgatttttag agtaaaaatc
     3541 actgatttta acacaaaatt ccagtttttc gtgatttttc actagaaaat cccgattttt
     3601 ccccaaaaaa tccatttttc cgactaaaaa tccccaaagt aattttcagg aggccaacaa
     3661 cgagacgtac gtcgagcaac acgaggtcgc cgtcgcccgt tactgcgacc tcccatccaa
     3721 gctcggacac aagctcaacg gtggcacagg aggaacgggc ggtagtggag gcactggtgc
     3781 cggcaaccca gctttcttgt acaaagttgg cattataaga aagcattgct tatcaatttg
     3841 ttgcaacgaa caggtcacta tcagtcaaaa taaaatcatt atttgccatc cagctgatat
     3901 cccctatagt gagtcgtatt acatggtcat agctgtttcc tggcagctct ggcccgtgtc
     3961 tcaaaatctc tgatgttaca ttgcacaaga taaaaatata tcatcatgaa caataaaact
     4021 gtctgcttac ataaacagta atacaagggg tgttatgagc catattcaac gggaaacgtc
     4081 gaggccgcga ttaaattcca acatggatgc tgatttatat gggtataaat gggctcgcga
     4141 taatgtcggg caatcaggtg cgacaatcta tcgcttgtat gggaagcccg atgcgccaga
     4201 gttgtttctg aaacatggca aaggtagcgt tgccaatgat gttacagatg agatggtcag
     4261 actaaactgg ctgacggaat ttatgcctct tccgaccatc aagcatttta tccgtactcc
     4321 tgatgatgca tggttactca ccactgcgat ccccggaaaa acagcattcc aggtattaga
     4381 agaatatcct gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt
     4441 gcattcgatt cctgtttgta attgtccttt taacagcgat cgcgtatttc gcctcgctca
     4501 ggcgcaatca cgaatgaata acggtttggt tgatgcgagt gattttgatg acgagcgtaa
     4561 tggctggcct gttgaacaag tctggaaaga aatgcataaa cttttgccat tctcaccgga
     4621 ttcagtcgtc actcatggtg atttctcact tgataacctt atttttgacg aggggaaatt
     4681 aataggttgt attgatgttg gacgagtcgg aatcgcagac cgataccagg atcttgccat
     4741 cctatggaac tgcctcggtg agttttctcc ttcattacag aaacggcttt ttcaaaaata
     4801 tggtattgat aatcctgata tgaataaatt gcagtttcat ttgatgctcg atgagttttt
     4861 ctaatcagaa ttggttaatt ggttgtaaca ctggcagagc attacgctga cttgacggga
     4921 cggcgcaagc tcatgaccaa aatcccttaa cgtgagttac gcgtcgttcc actgagcgtc
     4981 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     5041 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     5101 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     5161 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     5221 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     5281 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     5341 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     5401 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     5461 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     5521 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     5581 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     5641 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     5701 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
     5761 agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc
     5821 gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa
     5881 cgcaattaat acgcgtaccg ctagccagga agagtttgta gaaacgcaaa aaggccatcc
     5941 gtcaggatgg ccttctgctt agtttgatgc ctggcagttt atggcgggcg tcctgcccgc
     6001 caccctccgg gccgttgctt cacaacgttc aaatccgctc ccggcggatt tgtcctactc
     6061 aggagagcgt tcaccgacaa acaacagata aaacgaaagg cccagtcttc cgactgagcc
     6121 tttcgtttta tttgatgcct ggcagttccc tactctcgcg ttaacgctag catggatgtt
     6181 ttcccagtca cgacgtt
//
