LOCUS       sequence-319366-        3734 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PATC-rich intron number three from smu-2 to stimulate germline 
            expression in C. elegans.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3734)
  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 3734)
  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: pCFJ1149 - intron 3 smu-2 PATCs
FEATURES             Location/Qualifiers
     source          1..3734
                     /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(1859..1958)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1975..1994)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1976..1994)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1999..2015)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1999..2015)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2117..2136)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2128..2937
                     /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(2195..2214)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3084..3672
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3573..3592
                     /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 ttgaacggct gaaaaatcga tattttgagc gaaaaaagcc
      721 ggaaaaatgg attttatcgg ataaaatttg attttttgag ctgagaaatg cctttataga
      781 cgaatttcct tgtcaaattg ttaaaaactg caaaatttgc caaaaaaaaa gcatttttta
      841 tatttttttt tacaaaaaat tccacaaaaa tggcaaaatt tgactaaaaa tgccaatttt
      901 gtcgttttct ccgtgccaca gcggccgaaa atcgattttt aagcgatttt cgggtgaaaa
      961 agtgtaaaaa ccgactgaaa atccagctga aaacgacaaa aacggtaagt tttagctaat
     1021 tttcaccttt ttttcgatat ttttttacta aaaaaaaacc gaaaaattga gtttttttca
     1081 aatttctgca aaaattctgc aattttagca atttccatac tattttgacg ccgaaaaagc
     1141 ttaaaaatat gatttttaga ggtttttaat ggaaaattca tggatttttc aaatttttcc
     1201 acgaaaatca ttgaaaaatt tgaaaaaatt gatttcctat tcaaagttct agctaaaaac
     1261 tgcaatttta gcaatttcca tactatttta gtactgaaaa agcctaaaaa tgtgattttc
     1321 agaggatttc aatggaaaat tcatggattt ttccaatttt tcaatgaaaa ccattgaaaa
     1381 accctggaaa attgagtttt tcttcaaagt ttaggaaatt tatgagaatt tttgaataaa
     1441 aaatttttaa atttgaaaaa aattctcata aatttcgtaa acttttaata aaaactcaat
     1501 ttttccgatt tttttcgatg attttcacta aaaaattgga aaaattcatc aatttcctat
     1561 tgaaaacctc taaaaattat attttttttg cttttccgac gtcaaaatag tatggaaaat
     1621 gctaaaattg cagaattttt ccagaaattt gaagaaaaaa ctcaattttt tccgtttttt
     1681 caatgatttt catgaaaaaa ttggaaaatt ttgtaaattt tccattgaaa acctctgaaa
     1741 atttggaaat tttcgaattt aaatgtttaa aaaaaattga atattttgcg agtttttgaa
     1801 ttttttttga gaatttttga atttttttca cccaaaattt tttcaattag aagagaccac
     1861 ccagctttct tgtacaaagt tggcattata agaaagcatt gcttatcaat ttgttgcaac
     1921 gaacaggtca ctatcagtca aaataaaatc attatttgcc atccagctga tatcccctat
     1981 agtgagtcgt attacatggt catagctgtt tcctggcagc tctggcccgt gtctcaaaat
     2041 ctctgatgtt acattgcaca agataaaaat atatcatcat gaacaataaa actgtctgct
     2101 tacataaaca gtaatacaag gggtgttatg agccatattc aacgggaaac gtcgaggccg
     2161 cgattaaatt ccaacatgga tgctgattta tatgggtata aatgggctcg cgataatgtc
     2221 gggcaatcag gtgcgacaat ctatcgcttg tatgggaagc ccgatgcgcc agagttgttt
     2281 ctgaaacatg gcaaaggtag cgttgccaat gatgttacag atgagatggt cagactaaac
     2341 tggctgacgg aatttatgcc tcttccgacc atcaagcatt ttatccgtac tcctgatgat
     2401 gcatggttac tcaccactgc gatccccgga aaaacagcat tccaggtatt agaagaatat
     2461 cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg gttgcattcg
     2521 attcctgttt gtaattgtcc ttttaacagc gatcgcgtat ttcgtctcgc tcaggcgcaa
     2581 tcacgaatga ataacggttt ggttgatgcg agtgattttg atgacgagcg taatggctgg
     2641 cctgttgaac aagtctggaa agaaatgcat aaacttttgc cattctcacc ggattcagtc
     2701 gtcactcatg gtgatttctc acttgataac cttatttttg acgaggggaa attaataggt
     2761 tgtattgatg ttggacgagt cggaatcgca gaccgatacc aggatcttgc catcctatgg
     2821 aactgcctcg gtgagttttc tccttcatta cagaaacggc tttttcaaaa atatggtatt
     2881 gataatcctg atatgaataa attgcagttt catttgatgc tcgatgagtt tttctaatca
     2941 gaattggtta attggttgta acactggcag agcattacgc tgacttgacg ggacggcgca
     3001 agctcatgac caaaatccct taacgtgagt tacgcgtcgt tccactgagc gtcagacccc
     3061 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     3121 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     3181 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     3241 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     3301 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     3361 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     3421 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     3481 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     3541 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     3601 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     3661 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     3721 tttgctcaca tgtt
//
