LOCUS       sequence-317247-        6966 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Optimized enzyme for genome editing in C. elegans.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6966)
  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 6966)
  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: pMDJ21 - pEXP [Psmu-1 | eFlp recombinase (PATC...
FEATURES             Location/Qualifiers
     source          1..6966
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1..17
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    37..57
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     protein_bind    1050..1074
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3562..3586)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(4579..4599)
                     /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(4607..4624)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4607..4623)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4616..4638)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4951..4973
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5011..5029)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5097..5201
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5202..5270,5271..6062)
                     /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(5420..5439)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6233..6821
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6722..6741
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     join(6948..6966,1..4)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
ORIGIN
        1 caggaaacag ctatgaccat gattacgcca agctatcaac tttgtataga aaagttgaaa
       61 tttttcaggc gggaagtggc ttcggtgcgt tcaacaaacg cgcttttgac tcgctagccg
      121 gatccggctt cagcggtttt gacaaaagag ctttcgattc actggctgga caaggtttta
      181 ctggatttga gaaacgcgcc tttgatacag tggtacggtt ttcggtggaa attccagttt
      241 ttcgaatttt caataatttt cagagcacgt caggctttga cgatttcaaa ttgtgaagag
      301 ctcactgata tttaaaattt ggttcaattc aagcacatat ttttgaggta agcaactgat
      361 tttaatgttg ttaatcaaat gaaaagtgaa atttttggaa taattttttt ttgttttcag
      421 gtagctccaa cgaacgacac acacaagccc agtcccccag tattcctgac gacagcccct
      481 gctcacacac tttttattat aattcctttt ttttcaaact tttttttcaa attttatact
      541 ttaatctcaa aaaacctatt catacgaaca caagtaccgg tgtgcgtgtg tgagaataag
      601 aatttatttt aaaaaaataa agaatgtcgt cctgtttttt catatttatt gaagaaaatg
      661 gagctggaaa aaaatcaatt gtaaaaaaca gaaaaacgtg ctgataaaat gcattgtacg
      721 caaactgtgg attttgagaa ctaacgtagt caagcaaaaa tctctggaat tttatcgatt
      781 tttaacggat aaactgttgt tttttgtaac tgtgcatgaa aaaaaatctc aggcaaaaat
      841 attttctgaa aaaaaaagag tttaaacaca aatgcaagtc cacaaaaaac tccaatttga
      901 aattcgctcc aaaaacgcaa aacacgccaa gcgctcgtgc tggcgcattt gccgatcgct
      961 tccaaaaagg ctggcgcact tctcgtgtcc gaggatattc gaattttaca cgtcattttc
     1021 agccgttttt caccgaattt tcgctgaaaa caagtttgta caaaaaagca ggctaaaaat
     1081 gtcccaattc gacatcctct gcaaggtatt ttcctgcatt tttcaactgg gaaaatgaaa
     1141 gaaaatcgat aatttcagac cccaccaaag gtcctcgtcc gtcaattcgt cgagcgtttc
     1201 gagcgtccat ccggagagaa gatcgcctcc tgcgccgccg agctcaccta cctctgctgg
     1261 atgatcaccc acaacggaac cgccatcaag cgtgccacct tcatgtccta caacaccatc
     1321 atctccaact ccctctcctt cgacatcgtc aacaagtccc tccaattcaa gtacaaggta
     1381 agttttctta tgggaaagaa ggaaaaaacc gagattttac ttgaaaaatt gaatttttcg
     1441 cgggattttc accaaaaatt gttgaatatt cattatttca cgctgtaaaa caaaaaaaaa
     1501 aaaaatcaaa aactacgttg aaatcgcgtt tttaagcgaa ttttcttcag aattgccaga
     1561 ttttaacccc aaattttgca gtttttaaat aaaatttcac cttttcggct caaattgtag
     1621 attttcctga aaatttagta caaaaaacaa tttcctcgta aatttttcaa atagattttc
     1681 agacgcaaaa ggccaccatc ctcgaggcct ccctcaagaa gctcatccca gcctgggagt
     1741 tcaccatcat cccatacaac ggacaaaagc accaatccga catcaccgac atcgtctcct
     1801 ccctccaact ccaattcgag tcctccgagg aggccgacaa gggaaactcc cactccaaga
     1861 agatgctcaa ggccctcctc tccgagggag agtccatctg ggagatcacc gagaagatcc
     1921 tcaactcctt cgagtacacc tcccgtttca ccaagaccaa gaccctctac caattcctct
     1981 tcctcgccac cttcatcaac tgcggacgtt tctccgacat caaggtaagt tatgtctacc
     2041 tgcctgccta ccgcctaaat tttttgtgaa gtttcttcaa aaaatccaga aaaaaaaaca
     2101 attttcatac gattttttcc cttaaaattg tgaattttca tgctttttag ccccaaaagt
     2161 cattatttga gaaaaaattt catacaaaaa agttttgaga aatacacaat tttttaaatg
     2221 taattttcaa attttcaatt ttcaactaga aaattcacaa aacttgtaaa ttttggacca
     2281 aaacatttat acaattactt tttttgaatc taataactac aataactaca aatttaacat
     2341 tttcagaacg tcgacccaaa gtccttcaag ctcgtccaaa acaagtacct cggagtcatc
     2401 atccaatgcc tcgtcaccga gacgaagacc tccgtctccc gtcacatcta cttcttctcc
     2461 gcccgtggac gtatcgaccc actcgtctac ctcgacgagt tcctccgtaa ctccgagcca
     2521 gtcctcaagc gtgtcaaccg taccggaaac tcctcctcca acaagcaaga gtaccaactc
     2581 ctcaaggaca acctcgtccg ttcctacaac aaggccctca agaagaacgc cccataccca
     2641 atcttcgcca tcaaggtaag ttcttttttt gaaaagtgca gtttgtagtc taattttcat
     2701 ttattttctt tttaaaaaac gcatcaattt ttaaatattt tgggacaaaa tccgaaaact
     2761 gtactaattt gtagtttgta aaattaaaaa aaaaacgcaa aaaaatgttt tcaaaaatgc
     2821 ttagaataaa aataaaattt ctaaaaaatt gaacaaaata aacatttaaa aattcaaaaa
     2881 gtttgaaaaa atgcaaagtt ttttaatgca aaaaatattt aatgtttaaa caaatttaaa
     2941 aaatgttgta agaatatgtt tagagacagt ttttaaatgt taaattgcaa aaaaaaaaag
     3001 caaaaaaaaa tttgaaaaat gtaacagaaa attttaatta aaaaagtcgt gtttttttca
     3061 aacctgcaat tctttgtaat tttatttata aagagaaaca aatcaaattg tatattacga
     3121 aaaacgatac taaaattcga aaatttgcgg ttttttgcgt aaaaaatacg gttccgtaat
     3181 tttcagaacg gaccaaagtc ccacatcgga cgtcacctca tgacctcctt cctctccatg
     3241 aagggactca ccgagctcac caacgtcgtc ggaaactggt ccgacaagcg tgcctccgcc
     3301 gtcgcccgta ccacctacac ccaccaaatc accgccatcc cagaccacta cttcgccctc
     3361 gtctcccgtt actacgccta cgacccaatc tccaaggaga tgatcgccct caaggacgag
     3421 acaaacccaa tcgaggagtg gcaacacatc gagcaactca agggatccgc cgagggatcc
     3481 atccgttacc cagcctggaa cggaatcatc tcccaagagg tcctcgacta cctctcctcc
     3541 tacatcaacc gtcgtatcta gacccagctt tcttgtacaa agtggattaa ttttccccag
     3601 atttccccac gatttttttt gacattttta tgaaaatttt ctctaaaagt tacccctatt
     3661 ttgtttagac tgatctctta ttttatccaa tgtttcaaat ttttgaaatt gctcgaaaat
     3721 tccaattttc acatattttt gttgattttt tcgccatttt cccctgataa attaacttat
     3781 ttttgcaatt ttttctttcg aaaaaatatt ggtttctcat caccatttat cctttttttt
     3841 atggagttaa aaattgttaa aaaaatttaa ttttttttct aattttagag tcaaaacggg
     3901 ttcgaatcta gagcgtagct gtacgatttt ttggcgattt ttgcggtttt ctggtgagtt
     3961 tttttaaagt tatttttttg aaaaaaattc ttgaaaataa aaggaaattg ttttaaaaaa
     4021 aaatttcaaa gaaaaaattt tgaaaaaatt gaaacatttt ttttattttg accaaaaata
     4081 ttgaaaatta attaacattt ttttttggaa agttaatttt taatacaaat tttttaaatt
     4141 tattttacaa aaaaaattga atttctgtga aaaatcagtt ttccagtaca taatgatttg
     4201 accaaaaaaa attgaaaaaa atgatttttt aaaattaaaa ataccataaa atgctgaaaa
     4261 ttcagaaaaa taagtaaatt ttaaaaaatt ctggaaaaac atctaaaaac cccccaaata
     4321 taatattttg gaccctttta ttttctgctc gacatctcaa atatcctcca atttttcgct
     4381 tttttgatta attgaaagca tagtgggggc aggtgttata ggacaccctt ccctccactt
     4441 ttttggagca tttttcaaag ctcttccgag agcagaagct tctcaatttt tccaccccaa
     4501 aagtttgaaa ttctgccaaa aaatctcgca ttttaacaat ttttcgagat gtttcacgtc
     4561 cttttcattg cgatttccca actttattat acatagttga taattcactg gccgtcgttt
     4621 tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc
     4681 cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt
     4741 tgcgcagcct gaatggcgaa tggcgcctga tgcggtattt tctccttacg catctgtgcg
     4801 gtatttcaca ccgcatatgg tgcactctca gtgcaatctg ctctgatgcc gcatagttaa
     4861 gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg
     4921 catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac
     4981 cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta
     5041 atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg
     5101 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     5161 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     5221 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     5281 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     5341 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     5401 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     5461 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     5521 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     5581 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     5641 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     5701 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     5761 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     5821 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     5881 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     5941 tggggtcaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     6001 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     6061 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     6121 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     6181 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     6241 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     6301 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     6361 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     6421 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     6481 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     6541 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     6601 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     6661 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     6721 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     6781 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     6841 ttttacggtt cctggtcttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     6901 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccagc ggataacaat
     6961 ttcaca
//
