LOCUS       sequence-318053-        5432 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 5432)
  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 5432)
  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: pCFJ2211 - [1-2] ENTR - Fluor - mMaple3(intron...
FEATURES             Location/Qualifiers
     source          1..5432
                     /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(3021..3120)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3137..3156)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3138..3156)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3161..3177)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3161..3177)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3279..3298)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3290..4099
                     /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(3357..3376)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4246..4834
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4735..4754
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4988..5005
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(5164..5191)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(5283..5369)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     join(5418..5432,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(5432,1..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 ggaagcagtg
      181 tctccaaggt aacacttagt ttttgttgct ctgtttaaaa attaatttta agggagagga
      241 gactatcatg tccgtcatca agccagacat gaagatcaag ctccgtatgg agggaaacgt
      301 caacggacac gccttcgtca tcgagggaga gggatccgga aagccattcg agggaatcca
      361 aaccatcgac ctcgaggtca aggtaagttt tctgttgaac ggctgaaaaa tcgatatttt
      421 gagcgaaaaa agccggaaaa atggatttta tcggataaaa tttgattttt tgagctgaga
      481 aatgccttta tagacgaatt tccttgtcaa attgttaaaa actgcaaaat ttgccaaaaa
      541 aaaagcattt tttatatttt tttttacaaa aaattccaca aaaatggcaa aatttgacta
      601 aaaatgccaa ttttgtcgtt ttctccgtgc cacagcggcc gaaaatcgat ttttaagcga
      661 ttttcgggtg aaaaagtgta aaaaccgact gaaaatccag ctgaaaacga caaaaacggt
      721 aagttttagc taattttcac ctttttttcg atattttttt actaaaaaaa aaccgaaaaa
      781 ttgagttttt ttcaaatttc tgcaaaaatt ctgcaatttt agcaatttcc atactatttt
      841 gacgccgaaa aagcttaaaa atatgatttt tagaggtttt taatggaaaa ttcatggatt
      901 tttcaaattt ttccacgaaa atcattgaaa aatttgaaaa aattgatttc ctattcaaag
      961 ttctagctaa aaactgcaat tttagcaatt tccatactat tttagtactg aaaaagccta
     1021 aaaatgtgat tttcagagga tttcaatgga aaattcatgg atttttccaa tttttcaatg
     1081 aaaaccattg aaaaaccctg gaaaattgag tttttcttca aagtttagga aatttatgag
     1141 aatttttgaa taaaaaattt ttaaatttga aaaaaattct cataaatttc gtaaactttt
     1201 aataaaaact caatttttcc gatttttttc gatgattttc actaaaaaat tggaaaaatt
     1261 catcaatttc ctattgaaaa cctctaaaaa ttatattttt tttgcttttc cgacgtcaaa
     1321 atagtatgga aaatgctaaa attgcagaat ttttccagaa atttgaagaa aaaactcaat
     1381 tttttccgtt ttttcaatga ttttcatgaa aaaattggaa aattttgtaa attttccatt
     1441 gaaaacctct gaaaatttgg aaattttcga atttaaatgt ttaaaaaaaa ttgaatattt
     1501 tgcgagtttt tgaatttttt ttgagaattt ttgaattttt ttcacccaaa attttttcaa
     1561 ttagattttc aggagggagc cccactccca ttcgcctacg acatcctcac caccgccttc
     1621 cactacggaa accgtgtctt caccaagtac ccacgaaaga tcccagacta cttcaagcaa
     1681 tccttcccag gtaagttatg tctacctgcc tgcctaccgc ctaaattttt tgtgaagttt
     1741 cttcaaaaaa tccagaaaaa aaaacaattt tcatacgatt ttttccctta aaattgtgaa
     1801 ttttcatgct ttttagcccc aaaagtcatt atttgagaaa aaatttcata caaaaaagtt
     1861 ttgagaaata cacaattttt taaatgtaat tttcaaattt tcaattttca actagaaaat
     1921 tcacaaaact tgtaaatttt ggaccaaaac atttatacaa ttactttttt tgaatctaat
     1981 aactacaata actacaaatt taacattttc agagggatac tcctgggagc gttccatgac
     2041 ctacgaggac ggaggaatct gcaacgccac caacgacatc accatggagg aggactcctt
     2101 catcaacaag atccacttca agggaaccaa cttcccacca aacggaccag tcatgcaaaa
     2161 gcgtaccgtc ggatgggagg tttccaccga gaaggtaagt tctttttttg aaaagtgcag
     2221 tttgtagtct aattttcatt tattttcttt ttaaaaaacg catcaatttt taaatatttt
     2281 gggacaaaat ccgaaaactg tactaatttg tagtttgtaa aattaaaaaa aaaacgcaaa
     2341 aaaatgtttt caaaaatgct tagaataaaa ataaaatttc taaaaaattg aacaaaataa
     2401 acatttaaaa attcaaaaag tttgaaaaaa tgcaaagttt tttaatgcaa aaaatattta
     2461 atgtttaaac aaatttaaaa aatgttgtaa gaatatgttt agagacagtt tttaaatgtt
     2521 aaattgcaaa aaaaaaaagc aaaaaaaaat ttgaaaaatg taacagaaaa ttttaattaa
     2581 aaaagtcgtg tttttttcaa acctgcaatt ctttgtaatt ttatttataa agagaaacaa
     2641 atcaaattgt atattacgaa aaacgatact aaaattcgaa aatttgcggt tttttgcgta
     2701 aaaaatacgg ttccgtaatt ttcagatgta cgtccgtgac ggagtcctca agggtgacgt
     2761 caagatgaag ctcctcctca agggaggatc ccactaccgt tgcgacttcc gtaccaccta
     2821 caaggtcaag caaaaggccg tcaagctccc aaaggcccac ttcgtcgacc accgtatcga
     2881 gatcctctcc cacgacaagg actacaacaa ggtcaagctc tacgagcacg ccgtcgcccg
     2941 taactccacc gactccatgg acgagctcta caagggtggc acaggaggaa cgggcggtag
     3001 tggaggcact ggtgccggca acccagcttt cttgtacaaa gttggcatta taagaaagca
     3061 ttgcttatca atttgttgca acgaacaggt cactatcagt caaaataaaa tcattatttg
     3121 ccatccagct gatatcccct atagtgagtc gtattacatg gtcatagctg tttcctggca
     3181 gctctggccc gtgtctcaaa atctctgatg ttacattgca caagataaaa atatatcatc
     3241 atgaacaata aaactgtctg cttacataaa cagtaataca aggggtgtta tgagccatat
     3301 tcaacgggaa acgtcgaggc cgcgattaaa ttccaacatg gatgctgatt tatatgggta
     3361 taaatgggct cgcgataatg tcgggcaatc aggtgcgaca atctatcgct tgtatgggaa
     3421 gcccgatgcg ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca atgatgttac
     3481 agatgagatg gtcagactaa actggctgac ggaatttatg cctcttccga ccatcaagca
     3541 ttttatccgt actcctgatg atgcatggtt actcaccact gcgatccccg gaaaaacagc
     3601 attccaggta ttagaagaat atcctgattc aggtgaaaat attgttgatg cgctggcagt
     3661 gttcctgcgc cggttgcatt cgattcctgt ttgtaattgt ccttttaaca gcgatcgcgt
     3721 atttcgcctc gctcaggcgc aatcacgaat gaataacggt ttggttgatg cgagtgattt
     3781 tgatgacgag cgtaatggct ggcctgttga acaagtctgg aaagaaatgc ataaactttt
     3841 gccattctca ccggattcag tcgtcactca tggtgatttc tcacttgata accttatttt
     3901 tgacgagggg aaattaatag gttgtattga tgttggacga gtcggaatcg cagaccgata
     3961 ccaggatctt gccatcctat ggaactgcct cggtgagttt tctccttcat tacagaaacg
     4021 gctttttcaa aaatatggta ttgataatcc tgatatgaat aaattgcagt ttcatttgat
     4081 gctcgatgag tttttctaat cagaattggt taattggttg taacactggc agagcattac
     4141 gctgacttga cgggacggcg caagctcatg accaaaatcc cttaacgtga gttacgcgtc
     4201 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     4261 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     4321 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     4381 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     4441 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     4501 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     4561 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     4621 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     4681 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     4741 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     4801 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     4861 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     4921 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
     4981 cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct
     5041 ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc
     5101 gggcagtgag cgcaacgcaa ttaatacgcg taccgctagc caggaagagt ttgtagaaac
     5161 gcaaaaaggc catccgtcag gatggccttc tgcttagttt gatgcctggc agtttatggc
     5221 gggcgtcctg cccgccaccc tccgggccgt tgcttcacaa cgttcaaatc cgctcccggc
     5281 ggatttgtcc tactcaggag agcgttcacc gacaaacaac agataaaacg aaaggcccag
     5341 tcttccgact gagcctttcg ttttatttga tgcctggcag ttccctactc tcgcgttaac
     5401 gctagcatgg atgttttccc agtcacgacg tt
//
