LOCUS       sequence-319159-        3488 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Golden Gate compatible intron with PATCs for reduced germline 
            silencing.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3488)
  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 3488)
  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: pCFJ2215 - Intron GG3 - 900 bp
FEATURES             Location/Qualifiers
     source          1..3488
                     /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(1613..1712)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1729..1748)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1730..1748)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1753..1769)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1753..1769)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1871..1890)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1882..2691
                     /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(1949..1968)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2838..3426
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3327..3346
                     /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 agcaggctgg tctcactttt ttttgctcgt ttttgagtga aaaattgatg gaattcacaa
      721 aaaatggcca aagaatcctc aaaattggcc gaaaattctg aaaaatgacc taaaaatcac
      781 cgaaaaacgg ccaaaaatta tctgaaaatc gccgaaaatc tcaaattttg aggaaattcc
      841 cgctaaaaat taataaaaat caccgaaaaa aggccaaaaa ttacctaata aaggccgaaa
      901 atcgcctaaa aaggccaaaa attctgaaaa atggccaaat ttcaattttt cacacacaaa
      961 aaatccctga attttctatt tttactcgaa aaactgtgga tttacggggg taaaaattca
     1021 gtttttagtg ctatttcaca agaaaaacac agcattttcg acaaaaactc cgatttttaa
     1081 gccaaaaatt tgaggttttt agttttttac gtgaaaacct gaactttttg ccaaaaaaac
     1141 actttttttt agactaaaaa tcaattttcg atcaaaaatc actaattttg cctgaaaaaa
     1201 tatagatttt tccacttttt cactgaaaaa aaactggatt ttcactagaa aatcaagttg
     1261 tttttagacc aaaaatttta tttttccgct gaaaattatc gtttttcagt caattttcag
     1321 ctaaaaatcc ctgaaattct ggattttcag ccaaaaaact cgaaattttc caaaaaaacc
     1381 ccgtttaaaa atgagtttag actgaaaatt ccgatttttg agccaaaaat ttgattttta
     1441 atcgaaaaaa attggatttt caaccaaaaa atcagttttt cagtgatttt tagagtaaaa
     1501 atcactgatt ttaacacaaa attccagttt ttcgtgattt ttcactagaa aatcccgatt
     1561 tttccccaaa aaatccattt ttccgactaa aaatccccaa agtaaagaga ccacccagct
     1621 ttcttgtaca aagttggcat tataagaaag cattgcttat caatttgttg caacgaacag
     1681 gtcactatca gtcaaaataa aatcattatt tgccatccag ctgatatccc ctatagtgag
     1741 tcgtattaca tggtcatagc tgtttcctgg cagctctggc ccgtgtctca aaatctctga
     1801 tgttacattg cacaagataa aaatatatca tcatgaacaa taaaactgtc tgcttacata
     1861 aacagtaata caaggggtgt tatgagccat attcaacggg aaacgtcgag gccgcgatta
     1921 aattccaaca tggatgctga tttatatggg tataaatggg ctcgcgataa tgtcgggcaa
     1981 tcaggtgcga caatctatcg cttgtatggg aagcccgatg cgccagagtt gtttctgaaa
     2041 catggcaaag gtagcgttgc caatgatgtt acagatgaga tggtcagact aaactggctg
     2101 acggaattta tgcctcttcc gaccatcaag cattttatcc gtactcctga tgatgcatgg
     2161 ttactcacca ctgcgatccc cggaaaaaca gcattccagg tattagaaga atatcctgat
     2221 tcaggtgaaa atattgttga tgcgctggca gtgttcctgc gccggttgca ttcgattcct
     2281 gtttgtaatt gtccttttaa cagcgatcgc gtatttcgcc tcgctcaggc gcaatcacga
     2341 atgaataacg gtttggttga tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt
     2401 gaacaagtct ggaaagaaat gcataaactt ttgccattct caccggattc agtcgtcact
     2461 catggtgatt tctcacttga taaccttatt tttgacgagg ggaaattaat aggttgtatt
     2521 gatgttggac gagtcggaat cgcagaccga taccaggatc ttgccatcct atggaactgc
     2581 ctcggtgagt tttctccttc attacagaaa cggctttttc aaaaatatgg tattgataat
     2641 cctgatatga ataaattgca gtttcatttg atgctcgatg agtttttcta atcagaattg
     2701 gttaattggt tgtaacactg gcagagcatt acgctgactt gacgggacgg cgcaagctca
     2761 tgaccaaaat cccttaacgt gagttacgcg tcgttccact gagcgtcaga ccccgtagaa
     2821 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     2881 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     2941 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     3001 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     3061 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     3121 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     3181 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     3241 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     3301 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     3361 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     3421 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     3481 cacatgtt
//
