LOCUS       sequence-317103-        3169 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 3169)
  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 3169)
  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: pCFJ1359 - Intron GG2 - smu-1
FEATURES             Location/Qualifiers
     source          1..3169
                     /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(1365..1389)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        complement(1612..1630)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1613..1630)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     protein_bind    complement(1639..1659)
                     /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(1667..1684)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1667..1683)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1676..1698)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2011..2033
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2071..2089)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2157..2261
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2262..2330,2331..3122)
                     /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(2480..2499)
                     /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 ctggtctcaa tgtctacctg cctgcctacc gcctaaattt
     1081 tttgtgaagt ttcttcaaaa aatccagaaa aaaaaacaat tttcatacga ttttttccct
     1141 taaaattgtg aattttcatg ctttttagcc ccaaaagtca ttatttgaga aaaaatttca
     1201 tacaaaaaag ttttgagaaa tacacaattt tttaaatgta attttcaaat tttcaatttt
     1261 caactagaaa attcacaaaa cttgtaaatt ttggaccaaa acatttatac aattactttt
     1321 tttgaatcta ataactacaa taactacaaa tttaacaaga gaccacccag ctttcttgta
     1381 caaagtggac cagcctcaag aacacccgaa tggagtctct aagctacata ataccaactt
     1441 acactttaca aaatgttgtc ccccaaaatg tagccattcg tatctgctcc taataaaaag
     1501 aaagtttctt cacattctaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaam cccccccccc
     1561 cctgcaggca tgcattaatt aaaggccatg atggccagct agcttgagta ttctatagtg
     1621 tcacctaaat agcttggcca actttattat acatagttga taattcactg gccgtcgttt
     1681 tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc
     1741 cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt
     1801 tgcgcagcct gaatggcgaa tggcgcctga tgcggtattt tctccttacg catctgtgcg
     1861 gtatttcaca ccgcatatgg tgcactctca gtgcaatctg ctctgatgcc gcatagttaa
     1921 gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg
     1981 catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac
     2041 cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta
     2101 atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg
     2161 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     2221 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     2281 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     2341 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     2401 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     2461 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     2521 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     2581 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     2641 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     2701 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     2761 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     2821 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     2881 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     2941 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     3001 tggggtcaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     3061 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     3121 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaact
//
