LOCUS       sequence-319156-        3517 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 3517)
  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 3517)
  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: pCFJ2259 - Intron GG2 - 900 bp
FEATURES             Location/Qualifiers
     source          1..3517
                     /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(1642..1741)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1758..1777)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1759..1777)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1782..1798)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1782..1798)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1900..1919)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1911..2720
                     /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(1978..1997)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2867..3455
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3356..3375
                     /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 tctcaatgta cgcgatttct cggcgaaaaa atggactttt ttgctcaaaa
      721 actgaatttt tcagcttttt ttgcactgaa aatgctgaaa attggttttt ttgctgaaaa
      781 ttcaccgaaa aaagctgaaa atattgcaga aatcgacgaa aaatgccttt tttgctgaaa
      841 tttcagccag aaaattgttc agaatcgcaa tttcctgcca aaatcgacgt ttttcggcga
      901 aaattggtag attttgaacc gaaaattgtg aaaaatggac ttttttgctc aaaaacagac
      961 tttttgagct tttttcacac tgaaaatggt ggaaaattgg attttccagc agaaaattcg
     1021 gaaattttca atttttggcg gaaatttgac gaaaaatata tgaaaaacac cgatttttgt
     1081 ctgaaaaagt tcaaaagccg aaaaattggc caaaaatctc caaaatcgga cttttctggc
     1141 gtttttcgtg gaaaaattga aatttttttg aaaaattgag caaaaatcgc cgaaaaaccg
     1201 ctaaaaaaac tgccgaaatt ggcaattttg agcttaaaaa aacttgaaaa atgctcaaaa
     1261 ttacccaatt ttcaccggaa attgagctaa aatcgtcgaa aaagactgaa ttttcaattt
     1321 ttagctgaaa atctgcctat tttcggctaa aattgcttta aaaacattaa aattcaaatt
     1381 ttttggctaa aaatactcag attgcccttt tttttctcca aaaatgacaa aaaaaaaatc
     1441 aatcagaaaa atgcgatttt ttttacaaaa attggcttca aaacgtctaa attctcccta
     1501 atttttgctg ctgaaaattg tgcaaaaatc accgaaaaat gacgatttta ggctgaaaat
     1561 cttcaaaaaa agctcaaaat tccccacttt tcaccggaaa aagacgaaaa aagctccaaa
     1621 tttatcgatt aacaagagac cacccagctt tcttgtacaa agttggcatt ataagaaagc
     1681 attgcttatc aatttgttgc aacgaacagg tcactatcag tcaaaataaa atcattattt
     1741 gccatccagc tgatatcccc tatagtgagt cgtattacat ggtcatagct gtttcctggc
     1801 agctctggcc cgtgtctcaa aatctctgat gttacattgc acaagataaa aatatatcat
     1861 catgaacaat aaaactgtct gcttacataa acagtaatac aaggggtgtt atgagccata
     1921 ttcaacggga aacgtcgagg ccgcgattaa attccaacat ggatgctgat ttatatgggt
     1981 ataaatgggc tcgcgataat gtcgggcaat caggtgcgac aatctatcgc ttgtatggga
     2041 agcccgatgc gccagagttg tttctgaaac atggcaaagg tagcgttgcc aatgatgtta
     2101 cagatgagat ggtcagacta aactggctga cggaatttat gcctcttccg accatcaagc
     2161 attttatccg tactcctgat gatgcatggt tactcaccac tgcgatcccc ggaaaaacag
     2221 cattccaggt attagaagaa tatcctgatt caggtgaaaa tattgttgat gcgctggcag
     2281 tgttcctgcg ccggttgcat tcgattcctg tttgtaattg tccttttaac agcgatcgcg
     2341 tatttcgcct cgctcaggcg caatcacgaa tgaataacgg tttggttgat gcgagtgatt
     2401 ttgatgacga gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg cataaacttt
     2461 tgccattctc accggattca gtcgtcactc atggtgattt ctcacttgat aaccttattt
     2521 ttgacgaggg gaaattaata ggttgtattg atgttggacg agtcggaatc gcagaccgat
     2581 accaggatct tgccatccta tggaactgcc tcggtgagtt ttctccttca ttacagaaac
     2641 ggctttttca aaaatatggt attgataatc ctgatatgaa taaattgcag tttcatttga
     2701 tgctcgatga gtttttctaa tcagaattgg ttaattggtt gtaacactgg cagagcatta
     2761 cgctgacttg acgggacggc gcaagctcat gaccaaaatc ccttaacgtg agttacgcgt
     2821 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     2881 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     2941 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     3001 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     3061 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     3121 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     3181 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     3241 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     3301 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     3361 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     3421 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     3481 ggttcctggc cttttgctgg ccttttgctc acatgtt
//
