LOCUS       sequence-309304-        3565 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  piRNA-induced RNA interference (piRNAi) to induce males in 
            transgenic animals.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3565)
  AUTHORS   Priyadarshini M, Ni JZ, Vargas-Velazquez AM, Gu SG, Frokjaer-Jensen 
            C
  TITLE     Reprogramming the piRNA pathway for multiplexed and 
            transgenerational gene silencing in C. elegans.
  JOURNAL   Nat Methods. 2022 Feb;19(2):187-194. doi: 
            10.1038/s41592-021-01369-z. Epub 2022 Feb 3.
  PUBMED    35115715
REFERENCE   2  (bases 1 to 3565)
  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"; doi: 
            "10.1038/s41592-021-01369-z"; journalName: "Nat Methods"; date: 
            "2022-02"; volume: "19"; issue: "2"; pages: "187-194"
COMMENT     Sequence Label: pMNK54 - piRNAi 21ur-1224 - him-5
FEATURES             Location/Qualifiers
     source          1..3565
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(1418..1435)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1418..1434)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1427..1449)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        1548..1652
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1653..1721,1722..2513)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1871..1890)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     protein_bind    2613..2637
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     rep_origin      2754..3342
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3243..3262
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3495..3517
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3514..3530
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3514..3530
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 cactgagcct ccacctagcc tcgcgcttga cgcgctagtc aactaacata aaaaaggtga
       61 aacattgcga ggatacatag aaaaaacaat acttcgaatt catttttcaa ttacaaatcc
      121 tgaaatgttt cactgtgttc ctataagaaa acattgaaac aaaatattaa gtgagttagc
      181 tttccggagc ttctaatttt gattttgatt ttgaaatcga atttgcaaat ccaattaaaa
      241 atcattttct gataattaga cagttcctta tcgttaattt tattatatct atcgagttag
      301 aaattgcaac gaagataatg tcttccaaat actgaaaatt tgaaaatatg tttcctcacg
      361 aaaaacctgc ctattgccag aactcaaaat atgaaatttt tatagttttg ttgaaacagt
      421 aagaaaatct tgtaattact gtaaactgtt tgcttttttt aaagtcaacc tacttcaaat
      481 ctacttcaaa aattataatg tttcaaatta cataactgtg tatgcagaga gatcagtagg
      541 tactgtagag cttcaatgtt gataagattt attaacacag tgaaacaggt aatagttgtt
      601 tgttgcaaaa tcggaaatct ctacatttca tatggttttt aattacaggt ttgttttata
      661 aaataattgt gtgatggata ttattttcag acctcatact aatctgcaaa ccttcaaaca
      721 atatgtgaag tctactctgt ttcactcaac cattcatttc aatttggaaa aaaatcaaag
      781 aaatgttgaa aaattttcct gtttcaacat tatgacaaaa atgttatgat tttaataaaa
      841 acaatcgatc actgttgaca atcacttctg tttttcttag aagtgttttc cggaaacgcg
      901 taattggttt tatcacaaat cgaaaacaaa caaaaatttt tttaattatt tctttgctag
      961 ttttgtagtt gaaaattcac tataatcatg aataagtgag ctgcccaagt aaacaaagaa
     1021 aatttggcag cggccgacaa ctaccgggtt gcccgattta tcagtggagg ataatccggc
     1081 acgtagaatg tatctaatgt gatgtacacg gttttcattt aaaaacaaat tgaaacagaa
     1141 atgactacat tttcaaattg tctatttttg ctgtgtttat tttgccacca acaatcgatg
     1201 cgaccaactg ttttttcaat ctagtaaact cacttaatgc aattcctcca gccacatatg
     1261 taaacgttgt atacatgcag aaaacggttt tttggtttta atgggaactt ttgacaaatt
     1321 gttcgaaaat cttaagctgt cccatttcag ttgggtgatc gatttaggtc aggcggaatg
     1381 gcacttcctt attccagatg cgtgcggatt atgcatgact ggccgtcgtt ttacaacgtc
     1441 gtgactggga aaaccctggc gcttgtggtg gatcacagtt attggagact tccttggccg
     1501 tacgatgggt tttgtttcag gtatctggca cttttcgggg aaatgtgcgc ggaaccccta
     1561 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     1621 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     1681 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     1741 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     1801 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     1861 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     1921 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     1981 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     2041 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     2101 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     2161 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     2221 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     2281 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     2341 ctgataaatc tggagccggt gagcgtggct ctcgcggtat cattgcagca ctggggccag
     2401 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     2461 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     2521 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     2581 tctaggtgaa gatccttgaa ttgccattat caacaagttt gtacaaaaaa gcaggctccg
     2641 aattggtatc acgaggacga tttgaagccc cctacccatt tacgctgact tgacgggacg
     2701 gcggctttgt tgaatagagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     2761 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     2821 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     2881 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     2941 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     3001 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     3061 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     3121 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     3181 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     3241 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     3301 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     3361 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc agcgacatat
     3421 gccatacgcc gaactgaact ccacctaatg tcttggagat tgtcggatag cgcggaatag
     3481 gattgtgccc tctgagcgga taacaatttc acacaggaaa cagctatgac catgacacat
     3541 gcatccacca tcgcagactt atcat
//
