LOCUS       sequence-326626-        3454 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 3454)
  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 3454)
  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: pCFJ2214 - Intron GG1 - 900 bp
FEATURES             Location/Qualifiers
     source          1..3454
                     /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(1579..1678)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1695..1714)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1696..1714)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1719..1735)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1719..1735)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1837..1856)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1848..2657
                     /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(1915..1934)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2804..3392
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3293..3312
                     /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 tctcattctg ttttatttac ttgtttctga gaattgggat ttttagctga
      721 aaaattggat tttaagatga aaaattgaaa ttttcaacta aaaatttcaa tttttagctg
      781 aaaagttaaa tttttagcta aagagtggga tttttagttg aaaaatcgag atttctagct
      841 aaaaagtatg atttttagct gaaagattgg aattttaagc agaaaaaatt gtatttttag
      901 ctgataaatc aaattttaag cggaaaaatt tgaattttta gctaaaaaat tccattttca
      961 actaaaaatc gtgattttta gctttaaaaa ttggattttt agctgaaaaa tcggattttt
     1021 agctgaaaat tgaaattttt tagttaaaaa gtgagatttt ttaacagaaa agtgagattt
     1081 ttagttaaaa aataatattt ttagctaaaa aatacaattt tcaactaaag atcgtgattt
     1141 ttagctttaa aaattggatt tttagctaaa aaatcggatt tttagctgaa aattgaaatt
     1201 ttttagttaa aaagtgggat tttaagatga aaagtgagat tttttagctg aaaagtgaga
     1261 ttttcagcta aaaaatacaa ttttcaactg aaaaattggg atttttagct ttaaaaattg
     1321 gattttcagc tttaaaaatt ggatttttag ctgaaaagtg agatttttag ttgaaaaatt
     1381 gaaatgttta gctgaaaaat tggaattttt aagtaaaaaa gtgggatttt aagatgaaaa
     1441 gtgagatgtt tttagctgaa aagtgagatt tttagctaaa aaatacaatt ttcaactgaa
     1501 aaattgggat ttttagctca aaagtgggac gaaagacgaa aaaaaaaatc gattttttac
     1561 caaaaattag aagagaccac ccagctttct tgtacaaagt tggcattata agaaagcatt
     1621 gcttatcaat ttgttgcaac gaacaggtca ctatcagtca aaataaaatc attatttgcc
     1681 atccagctga tatcccctat agtgagtcgt attacatggt catagctgtt tcctggcagc
     1741 tctggcccgt gtctcaaaat ctctgatgtt acattgcaca agataaaaat atatcatcat
     1801 gaacaataaa actgtctgct tacataaaca gtaatacaag gggtgttatg agccatattc
     1861 aacgggaaac gtcgaggccg cgattaaatt ccaacatgga tgctgattta tatgggtata
     1921 aatgggctcg cgataatgtc gggcaatcag gtgcgacaat ctatcgcttg tatgggaagc
     1981 ccgatgcgcc agagttgttt ctgaaacatg gcaaaggtag cgttgccaat gatgttacag
     2041 atgagatggt cagactaaac tggctgacgg aatttatgcc tcttccgacc atcaagcatt
     2101 ttatccgtac tcctgatgat gcatggttac tcaccactgc gatccccgga aaaacagcat
     2161 tccaggtatt agaagaatat cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt
     2221 tcctgcgccg gttgcattcg attcctgttt gtaattgtcc ttttaacagc gatcgcgtat
     2281 ttcgcctcgc tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg agtgattttg
     2341 atgacgagcg taatggctgg cctgttgaac aagtctggaa agaaatgcat aaacttttgc
     2401 cattctcacc ggattcagtc gtcactcatg gtgatttctc acttgataac cttatttttg
     2461 acgaggggaa attaataggt tgtattgatg ttggacgagt cggaatcgca gaccgatacc
     2521 aggatcttgc catcctatgg aactgcctcg gtgagttttc tccttcatta cagaaacggc
     2581 tttttcaaaa atatggtatt gataatcctg atatgaataa attgcagttt catttgatgc
     2641 tcgatgagtt tttctaatca gaattggtta attggttgta acactggcag agcattacgc
     2701 tgacttgacg ggacggcgca agctcatgac caaaatccct taacgtgagt tacgcgtcgt
     2761 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     2821 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     2881 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     2941 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     3001 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     3061 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     3121 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     3181 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     3241 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     3301 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     3361 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     3421 tcctggcctt ttgctggcct tttgctcaca tgtt
//
