LOCUS       sequence-317110-        3376 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 3376)
  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 3376)
  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: pCFJ1360 - Intron GG3 - smu-1
FEATURES             Location/Qualifiers
     source          1..3376
                     /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(1574..1598)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        complement(1819..1837)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1820..1837)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     protein_bind    complement(1846..1866)
                     /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(1874..1891)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1874..1890)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1883..1905)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2218..2240
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2278..2296)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2364..2468
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2469..2537,2538..3329)
                     /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(2687..2706)
                     /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 ctggtctcac tttttttgaa aagtgcagtt tgtagtctaa
     1081 ttttcattta ttttcttttt aaaaaacgca tcaattttta aatattttgg gacaaaatcc
     1141 gaaaactgta ctaatttgta gtttgtaaaa ttaaaaaaaa aacgcaaaaa aatgttttca
     1201 aaaatgctta gaataaaaat aaaatttcta aaaaattgaa caaaataaac atttaaaaat
     1261 tcaaaaagtt tgaaaaaatg caaagttttt taatgcaaaa aatatttaat gtttaaacaa
     1321 atttaaaaaa tgttgtaaga atatgtttag agacagtttt taaatgttaa attgcaaaaa
     1381 aaaaaagcaa aaaaaaattt gaaaaatgta acagaaaatt ttaattaaaa aagtcgtgtt
     1441 tttttcaaac ctgcaattct ttgtaatttt atttataaag agaaacaaat caaattgtat
     1501 attacgaaaa acgatactaa aattcgaaaa tttgcggttt tttgcgtaaa aaatacggtt
     1561 ccgtaaagag accacccagc tttcttgtac aaagtggacc agcctcaaga acacccgaat
     1621 ggagtctcta agctacataa taccaactta cactttacaa aatgttgtcc cccaaaatgt
     1681 agccattcgt atctgctcct aataaaaaga aagtttcttc acattctaaa aaaaaaaaaa
     1741 aaaaaaaaaa aaaaaaamcc ccccccccct gcaggcatgc attaattaaa ggccatgatg
     1801 gccagctagc ttgagtattc tatagtgtca cctaaatagc ttggccaact ttattataca
     1861 tagttgataa ttcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta
     1921 cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg
     1981 cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg cgcctgatgc
     2041 ggtattttct ccttacgcat ctgtgcggta tttcacaccg catatggtgc actctcagtg
     2101 caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg
     2161 cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg
     2221 ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc
     2281 tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag
     2341 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     2401 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     2461 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     2521 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     2581 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     2641 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     2701 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     2761 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     2821 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     2881 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     2941 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     3001 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     3061 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     3121 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     3181 gtgggtctcg cggtatcatt gcagcactgg ggtcagatgg taagccctcc cgtatcgtag
     3241 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     3301 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     3361 agattgattt aaaact
//
