LOCUS       sequence-319163-        2816 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Golden Gate compatible intron with no PATCs for negative controls.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2816)
  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 2816)
  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: pCFJ2366 - intron GG3 - 250bp no PATC
FEATURES             Location/Qualifiers
     source          1..2816
                     /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(941..1040)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1057..1076)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1058..1076)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1081..1097)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1081..1097)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1199..1218)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1210..2019
                     /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(1277..1296)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2166..2754
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2655..2674
                     /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 tctcacttta tgaagatacg aattaaaaaa aagacgacgc ctcttcatca
      721 cttcaatgta acaaagtgcc tttttgaacg cacacacttc ttccccacgt gctgttgcag
      781 ttttgatact aaaaaaacga tagaaaaaga gagataagag atgaaaatga taagagaatt
      841 ggaaaaagag gaattaaaaa aggaaacgat taagttgaaa gtagaaagaa cagaaaaatg
      901 tgtatggagg ggttttcaaa ttctcttttg taaagagacc acccagcttt cttgtacaaa
      961 gttggcatta taagaaagca ttgcttatca atttgttgca acgaacaggt cactatcagt
     1021 caaaataaaa tcattatttg ccatccagct gatatcccct atagtgagtc gtattacatg
     1081 gtcatagctg tttcctggca gctctggccc gtgtctcaaa atctctgatg ttacattgca
     1141 caagataaaa atatatcatc atgaacaata aaactgtctg cttacataaa cagtaataca
     1201 aggggtgtta tgagccatat tcaacgggaa acgtcgaggc cgcgattaaa ttccaacatg
     1261 gatgctgatt tatatgggta taaatgggct cgcgataatg tcgggcaatc aggtgcgaca
     1321 atctatcgct tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca tggcaaaggt
     1381 agcgttgcca atgatgttac agatgagatg gtcagactaa actggctgac ggaatttatg
     1441 cctcttccga ccatcaagca ttttatccgt actcctgatg atgcatggtt actcaccact
     1501 gcgatccccg gaaaaacagc attccaggta ttagaagaat atcctgattc aggtgaaaat
     1561 attgttgatg cgctggcagt gttcctgcgc cggttgcatt cgattcctgt ttgtaattgt
     1621 ccttttaaca gcgatcgcgt atttcgcctc gctcaggcgc aatcacgaat gaataacggt
     1681 ttggttgatg cgagtgattt tgatgacgag cgtaatggct ggcctgttga acaagtctgg
     1741 aaagaaatgc ataaactttt gccattctca ccggattcag tcgtcactca tggtgatttc
     1801 tcacttgata accttatttt tgacgagggg aaattaatag gttgtattga tgttggacga
     1861 gtcggaatcg cagaccgata ccaggatctt gccatcctat ggaactgcct cggtgagttt
     1921 tctccttcat tacagaaacg gctttttcaa aaatatggta ttgataatcc tgatatgaat
     1981 aaattgcagt ttcatttgat gctcgatgag tttttctaat cagaattggt taattggttg
     2041 taacactggc agagcattac gctgacttga cgggacggcg caagctcatg accaaaatcc
     2101 cttaacgtga gttacgcgtc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     2161 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     2221 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     2281 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     2341 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     2401 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     2461 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     2521 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     2581 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     2641 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     2701 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     2761 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgtt
//
