LOCUS       sequence-318007-        6218 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 6218)
  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 6218)
  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: pCFJ2440 - [1-2] ENTR - Fluor - ce-GFP(PATCs(9...
FEATURES             Location/Qualifiers
     source          1..6218
                     /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(3807..3906)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3923..3942)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3924..3942)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3947..3963)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3947..3963)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4065..4084)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4076..4885
                     /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(4143..4162)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      5032..5620
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5521..5540
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5774..5791
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(5950..5977)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(6069..6155)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     join(6204..6218,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(6218,1..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 attgataagc aatgctttct tataatgcca actttgtaca
      121 aaaaagcagg ctcggccggc agtactagcg gtggcagtgg aggtaccggc ggaagcagtt
      181 ccaaggtaac acttagtttt tgttgctctg tttaaaaatt aattttaagg gagaggagct
      241 gttcaccgga gtcgtcccaa tcctcgtcga gctcgacggg gacgtcaacg gacacaagtt
      301 ctccgtgtcc ggagagggag agggagatgc cacctacgga aagctcaccc tcaagttcat
      361 ctgcaccacc ggaaagctcc cagtcccatg gccaaccctc gtcaccacct tctgctacgg
      421 agtccaatgc ttctcccgtt acccaggtaa gttttctgtt ttatttactt gttttgagaa
      481 ttgggatttt tagctgaaaa attggatttt aagatgaaaa attgaaattt tcaactaaaa
      541 atttcaattt ttagctgaaa agttaaattt ttagctaaag agtgggattt ttagttgaaa
      601 aatcgagatt tctagctaaa aagtatgatt tttagctgaa agattggaat tttaagcaga
      661 aaaaattgta tttttagctg ataaatcaaa ttttaagcgg aaaaatttga atttttagct
      721 aaaaaattcc attttcaact aaaaatcgtg atttttagct ttaaaaattg gatttttagc
      781 tgaaaaatcg gatttttagc tgaaaattga aattttttag ttaaaaagtg agatttttta
      841 acagaaaagt gagattttta gttaaaaaat aatattttta gctaaaaaat acaattttca
      901 actaaagatc gtgattttta gctttaaaaa ttggattttt agctaaaaaa tcggattttt
      961 agctgaaaat tgaaattttt tagttaaaaa gtgggatttt aagatgaaaa gtgagatttt
     1021 ttagctgaaa agtgagattt tcagctaaaa aatacaattt tcaactgaaa aattgggatt
     1081 tttagcttta aaaattggat tttcagcttt aaaaattgga tttttagctg aaaagtgaga
     1141 tttttagttg aaaaattgaa atgtttagct gaaaaattgg aatttttaag taaaaaagtg
     1201 ggattttaag atgaaaagtg agatgttttt agctgaaaag tgagattttt agctaaaaaa
     1261 tacaattttc aactgaaaaa ttgggatttt tagctcaaaa gtgggacgaa agacgaaaaa
     1321 aaaaatcgat tttttaccaa aaattagatt ttcagaccac atgaagcgtc acgacttctt
     1381 caagtccgcc atgccagagg gatacgtcca agagcgtacc atcttcttca aggacgacgg
     1441 aaactacaag gtaagttatg tacgcgattt ctcggcgaaa aaatggactt ttttgctcaa
     1501 aaactgaatt tttcagcttt ttttgcactg aaaatgctga aaattggttt ttttgctgaa
     1561 aattcaccga aaaaagctga aaatattgca gaaatcgacg aaaaatgcct tttttgctga
     1621 aatttcagcc agaaaattgt tcagaatcgc aatttcctgc caaaatcgac gtttttcggc
     1681 gaaaattggt agattttgaa ccgaaaattg tgaaaaatgg acttttttgc tcaaaaacag
     1741 actttttgag cttttttcac actgaaaatg gtggaaaatt ggattttcca gcagaaaatt
     1801 cggaaatttt caatttttgg cggaaatttg acgaaaaata tatgaaaaac accgattttt
     1861 gtctgaaaaa gttcaaaagc cgaaaaattg gccaaaaatc tccaaaatcg gacttttctg
     1921 gcgtttttcg tggaaaaatt gaaatttttt tgaaaaattg agcaaaaatc gccgaaaaac
     1981 cgctaaaaaa actgccgaaa ttggcaattt tgagcttaaa aaaacttgaa aaatgctcaa
     2041 aattacccaa ttttcaccgg aaattgagct aaaatcgtcg aaaaagactg aattttcaat
     2101 ttttagctga aaatctgcct attttcggct aaaattgctt taaaaacatt aaaattcaaa
     2161 ttttttggct aaaaatactc agattgccct tttttttctc caaaaatgac aaaaaaaaaa
     2221 tcaatcagaa aaatgcgatt ttttttacaa aaattggctt caaaacgtct aaattctccc
     2281 taatttttgc tgctgaaaat tgtgcaaaaa tcaccgaaaa atgacgattt taggctgaaa
     2341 atcttcaaaa aaagctcaaa attccccact tttcaccgga aaaagacgaa aaaagctcca
     2401 aatttatcga ttaacatttt cagacccgtg ccgaggtcaa gttcgaggga gacaccctcg
     2461 tcaaccgtat cgagctcaag ggaatcgact tcaaggagga cggtaacatc ctgggacaca
     2521 agctggagta caactacaac tcccacaacg tctacatcat ggccgacaag caaaagaacg
     2581 gaatcaaggt caacttcaag gtaagttctt ttttttgctc gtttttgagt gaaaaattga
     2641 tggaattcac aaaaaatggc caaagaatcc tcaaaattgg ccgaaaattc tgaaaaatga
     2701 cctaaaaatc accgaaaaac ggccaaaaat tatctgaaaa tcgccgaaaa tctcaaattt
     2761 tgaggaaatt cccgctaaaa attaataaaa atcaccgaaa aaaggccaaa aattacctaa
     2821 taaaggccga aaatcgccta aaaaggccaa aaattctgaa aaatggccaa atttcaattt
     2881 ttcacacaca aaaaatccct gaattttcta tttttactcg aaaaactgtg gatttacggg
     2941 ggtaaaaatt cagtttttag tgctatttca caagaaaaac acagcatttt cgacaaaaac
     3001 tccgattttt aagccaaaaa tttgaggttt ttagtttttt acgtgaaaac ctgaactttt
     3061 tgccaaaaaa acactttttt ttagactaaa aatcaatttt cgatcaaaaa tcactaattt
     3121 tgcctgaaaa aatatagatt tttccacttt ttcactgaaa aaaaactgga ttttcactag
     3181 aaaatcaagt tgtttttaga ccaaaaattt tatttttccg ctgaaaatta tcgtttttca
     3241 gtcaattttc agctaaaaat ccctgaaatt ctggattttc agccaaaaaa ctcgaaattt
     3301 tccaaaaaaa ccccgtttaa aaatgagttt agactgaaaa ttccgatttt tgagccaaaa
     3361 atttgatttt taatcgaaaa aaattggatt ttcaaccaaa aaatcagttt ttcagtgatt
     3421 tttagagtaa aaatcactga ttttaacaca aaattccagt ttttcgtgat ttttcactag
     3481 aaaatcccga tttttcccca aaaaatccat ttttccgact aaaaatcccc aaagtaattt
     3541 tcagatccgt cacaacatcg aggacggatc ggtccaactc gccgaccact accaacaaaa
     3601 caccccaatc ggtgacggac cagtcctcct cccagacaac cactacctct ccacccaatc
     3661 cgccctctcc aaggacccaa acgagaagcg tgaccacatg gtcctcctcg agttcgtcac
     3721 cgccgccgga atcacccacg gaatggacga gctctacaag ggtggcacag gaggaacggg
     3781 cggtagtgga ggcactggtg ccggcaaccc agctttcttg tacaaagttg gcattataag
     3841 aaagcattgc ttatcaattt gttgcaacga acaggtcact atcagtcaaa ataaaatcat
     3901 tatttgccat ccagctgata tcccctatag tgagtcgtat tacatggtca tagctgtttc
     3961 ctggcagctc tggcccgtgt ctcaaaatct ctgatgttac attgcacaag ataaaaatat
     4021 atcatcatga acaataaaac tgtctgctta cataaacagt aatacaaggg gtgttatgag
     4081 ccatattcaa cgggaaacgt cgaggccgcg attaaattcc aacatggatg ctgatttata
     4141 tgggtataaa tgggctcgcg ataatgtcgg gcaatcaggt gcgacaatct atcgcttgta
     4201 tgggaagccc gatgcgccag agttgtttct gaaacatggc aaaggtagcg ttgccaatga
     4261 tgttacagat gagatggtca gactaaactg gctgacggaa tttatgcctc ttccgaccat
     4321 caagcatttt atccgtactc ctgatgatgc atggttactc accactgcga tccccggaaa
     4381 aacagcattc caggtattag aagaatatcc tgattcaggt gaaaatattg ttgatgcgct
     4441 ggcagtgttc ctgcgccggt tgcattcgat tcctgtttgt aattgtcctt ttaacagcga
     4501 tcgcgtattt cgcctcgctc aggcgcaatc acgaatgaat aacggtttgg ttgatgcgag
     4561 tgattttgat gacgagcgta atggctggcc tgttgaacaa gtctggaaag aaatgcataa
     4621 acttttgcca ttctcaccgg attcagtcgt cactcatggt gatttctcac ttgataacct
     4681 tatttttgac gaggggaaat taataggttg tattgatgtt ggacgagtcg gaatcgcaga
     4741 ccgataccag gatcttgcca tcctatggaa ctgcctcggt gagttttctc cttcattaca
     4801 gaaacggctt tttcaaaaat atggtattga taatcctgat atgaataaat tgcagtttca
     4861 tttgatgctc gatgagtttt tctaatcaga attggttaat tggttgtaac actggcagag
     4921 cattacgctg acttgacggg acggcgcaag ctcatgacca aaatccctta acgtgagtta
     4981 cgcgtcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     5041 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     5101 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     5161 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     5221 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     5281 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     5341 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     5401 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     5461 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     5521 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     5581 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     5641 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     5701 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
     5761 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc
     5821 gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg
     5881 gaaagcgggc agtgagcgca acgcaattaa tacgcgtacc gctagccagg aagagtttgt
     5941 agaaacgcaa aaaggccatc cgtcaggatg gccttctgct tagtttgatg cctggcagtt
     6001 tatggcgggc gtcctgcccg ccaccctccg ggccgttgct tcacaacgtt caaatccgct
     6061 cccggcggat ttgtcctact caggagagcg ttcaccgaca aacaacagat aaaacgaaag
     6121 gcccagtctt ccgactgagc ctttcgtttt atttgatgcc tggcagttcc ctactctcgc
     6181 gttaacgcta gcatggatgt tttcccagtc acgacgtt
//
