LOCUS       sequence-316190-        3152 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 3152)
  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 3152)
  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: pCFJ1358 - Intron GG1 - smu-1
FEATURES             Location/Qualifiers
     source          1..3152
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      124..712
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     613..632
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     839..861
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     858..874
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     858..874
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    894..914
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     primer_bind     923..942
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        923..941
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     5'UTR           973..1015
                     /label=Xenopus globin 5'-UTR
                     /note="translational enhancer from the 5'-UTR of the major 
                     beta-globin gene of Xenopus laevis"
     protein_bind    1018..1042
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(1348..1372)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        complement(1595..1613)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1596..1613)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     protein_bind    complement(1622..1642)
                     /gene="mutant version of attB"
                     /label=attB3
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     primer_bind     complement(1650..1667)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1650..1666)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1659..1681)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1994..2016
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2054..2072)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2140..2244
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2245..2313,2314..3105)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGSDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2463..2482)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
       61 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
      121 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
      181 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
      241 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
      301 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
      361 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
      421 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
      481 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
      541 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
      601 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
      661 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
      721 caacgcggcc tttttacggt tcctggtctt ttgctggcct tttgctcaca tgttctttcc
      781 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccag
      841 cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc tatcaacttt
      901 gtatagaaaa gttggtgaat tgtaatacga ctcactatag ggcgaattaa ttcgagctcg
      961 gtacccagct tgcttgttct ttttgcagaa gctcagaata aacgctcaac tttggcagca
     1021 agtttgtaca aaaaagcagg ctggtctcat tcttatggga aagaaggaaa aaaccgagat
     1081 tttacttgaa aaattgaatt tttcgcggga ttttcaccaa aaattgttga atattcatta
     1141 tttcacgctg taaaacaaaa aaaaaaaaaa tcaaaaacta cgttgaaatc gcgtttttaa
     1201 gcgaattttc ttcagaattg ccagatttta accccaaatt ttgcagtttt taaataaaat
     1261 ttcacctttt cggctcaaat tgtagatttt cctgaaaatt tagtacaaaa aacaatttcc
     1321 tcgtaaattt ttcaaataga agagaccacc cagctttctt gtacaaagtg gaccagcctc
     1381 aagaacaccc gaatggagtc tctaagctac ataataccaa cttacacttt acaaaatgtt
     1441 gtcccccaaa atgtagccat tcgtatctgc tcctaataaa aagaaagttt cttcacattc
     1501 taaaaaaaaa aaaaaaaaaa aaaaaaaaaa aamccccccc ccccctgcag gcatgcatta
     1561 attaaaggcc atgatggcca gctagcttga gtattctata gtgtcaccta aatagcttgg
     1621 ccaactttat tatacatagt tgataattca ctggccgtcg ttttacaacg tcgtgactgg
     1681 gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg
     1741 cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc
     1801 gaatggcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata
     1861 tggtgcactc tcagtgcaat ctgctctgat gccgcatagt taagccagcc ccgacacccg
     1921 ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa
     1981 gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc
     2041 gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg
     2101 gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     2161 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     2221 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     2281 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     2341 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     2401 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     2461 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
     2521 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     2581 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     2641 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     2701 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     2761 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
     2821 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     2881 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     2941 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggtc agatggtaag
     3001 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     3061 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     3121 tactcatata tactttagat tgatttaaaa ct
//
