LOCUS       sequence-319103-        4382 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Three-fragment Gateway compatible protein tag for expression in C. 
            elegans.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4382)
  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 4382)
  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: pCFJ2089 - [1-2] ENTR - Tag - SNAP(PATC, no_at...
FEATURES             Location/Qualifiers
     source          1..4382
                     /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(2507..2606)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2623..2642)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2624..2642)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2647..2663)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2647..2663)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2765..2784)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2776..3585
                     /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(2843..2862)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3732..4320
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4221..4240
                     /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 gcagtgacaa
      721 ggtaacactt agtttttgtt gctctgttta aaaattaatt ttaaggactg cgagatgaag
      781 cgtaccaccc tcgactcccc actcggaaag ctcgagctct ccggatgcga gcaaggactc
      841 caccgtatca tcttcctcgg aaagggaacc tccgccgccg acgccgtcga ggtcccagcc
      901 ccagccgccg tcctcggagg accaggtaag ttttcttatg ggaaagaagg aaaaaaccga
      961 gattttactt gaaaaattga atttttcgcg ggattttcac caaaaattgt tgaatattca
     1021 ttatttcacg ctgtaaaaca aaaaaaaaaa aaatcaaaaa ctacgttgaa atcgcgtttt
     1081 taagcgaatt ttcttcagaa ttgccagatt ttaaccccaa attttgcagt ttttaaataa
     1141 aatttcacct tttcggctca aattgtagat tttcctgaaa atttagtaca aaaaacaatt
     1201 tcctcgtaaa tttttcaaat agattttcag agccactcat gcaagccacc gcctggctca
     1261 acgcctactt ccaccaacca gaggccatcg aggagttccc agtcccagcc ctccaccacc
     1321 cagtcttcca acaagagtcc ttcacccgtc aagtcctctg gaagctcctc aaggtaagtt
     1381 atgtctacct gcctgcctac cgcctaaatt ttttgtgaag tttcttcaaa aaatccagaa
     1441 aaaaaaacaa ttttcatacg attttttccc ttaaaattgt gaattttcat gctttttagc
     1501 cccaaaagtc attatttgag aaaaaatttc atacaaaaaa gttttgagaa atacacaatt
     1561 ttttaaatgt aattttcaaa ttttcaattt tcaactagaa aattcacaaa acttgtaaat
     1621 tttggaccaa aacatttata caattacttt ttttgaatct aataactaca ataactacaa
     1681 atttaacatt ttcaggtcgt caagttcgga gaggtcatct cctactccca cctcgccgcc
     1741 ctcgccggaa acccagccgc caccgccgcc gtcaagaccg ccctctccgg aaacccagtc
     1801 ccaatcctca tcccatgcca ccgtgtcgtc caaggagacc tcgacgtcgg aggatacgag
     1861 ggaggactcg ccgtcaaggt aagttctttt tttgaaaagt gcagtttgta gtctaatttt
     1921 catttatttt ctttttaaaa aacgcatcaa tttttaaata ttttgggaca aaatccgaaa
     1981 actgtactaa tttgtagttt gtaaaattaa aaaaaaaacg caaaaaaatg ttttcaaaaa
     2041 tgcttagaat aaaaataaaa tttctaaaaa attgaacaaa ataaacattt aaaaattcaa
     2101 aaagtttgaa aaaatgcaaa gttttttaat gcaaaaaata tttaatgttt aaacaaattt
     2161 aaaaaatgtt gtaagaatat gtttagagac agtttttaaa tgttaaattg caaaaaaaaa
     2221 aagcaaaaaa aaatttgaaa aatgtaacag aaaattttaa ttaaaaaagt cgtgtttttt
     2281 tcaaacctgc aattctttgt aattttattt ataaagagaa acaaatcaaa ttgtatatta
     2341 cgaaaaacga tactaaaatt cgaaaatttg cggttttttg cgtaaaaaat acggttccgt
     2401 aattttcagg agtggctcct cgcccacgag ggacaccctc tcggaaagcc aggactcgga
     2461 ggtggcacag gaggaacggg cggtagtgga ggcactggtg ccggcaaccc agctttcttg
     2521 tacaaagttg gcattataag aaagcattgc ttatcaattt gttgcaacga acaggtcact
     2581 atcagtcaaa ataaaatcat tatttgccat ccagctgata tcccctatag tgagtcgtat
     2641 tacatggtca tagctgtttc ctggcagctc tggcccgtgt ctcaaaatct ctgatgttac
     2701 attgcacaag ataaaaatat atcatcatga acaataaaac tgtctgctta cataaacagt
     2761 aatacaaggg gtgttatgag ccatattcaa cgggaaacgt cgaggccgcg attaaattcc
     2821 aacatggatg ctgatttata tgggtataaa tgggctcgcg ataatgtcgg gcaatcaggt
     2881 gcgacaatct atcgcttgta tgggaagccc gatgcgccag agttgtttct gaaacatggc
     2941 aaaggtagcg ttgccaatga tgttacagat gagatggtca gactaaactg gctgacggaa
     3001 tttatgcctc ttccgaccat caagcatttt atccgtactc ctgatgatgc atggttactc
     3061 accactgcga tccccggaaa aacagcattc caggtattag aagaatatcc tgattcaggt
     3121 gaaaatattg ttgatgcgct ggcagtgttc ctgcgccggt tgcattcgat tcctgtttgt
     3181 aattgtcctt ttaacagcga tcgcgtattt cgcctcgctc aggcgcaatc acgaatgaat
     3241 aacggtttgg ttgatgcgag tgattttgat gacgagcgta atggctggcc tgttgaacaa
     3301 gtctggaaag aaatgcataa acttttgcca ttctcaccgg attcagtcgt cactcatggt
     3361 gatttctcac ttgataacct tatttttgac gaggggaaat taataggttg tattgatgtt
     3421 ggacgagtcg gaatcgcaga ccgataccag gatcttgcca tcctatggaa ctgcctcggt
     3481 gagttttctc cttcattaca gaaacggctt tttcaaaaat atggtattga taatcctgat
     3541 atgaataaat tgcagtttca tttgatgctc gatgagtttt tctaatcaga attggttaat
     3601 tggttgtaac actggcagag cattacgctg acttgacggg acggcgcaag ctcatgacca
     3661 aaatccctta acgtgagtta cgcgtcgttc cactgagcgt cagaccccgt agaaaagatc
     3721 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     3781 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     3841 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     3901 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     3961 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     4021 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     4081 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     4141 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     4201 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     4261 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     4321 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     4381 tt
//
