LOCUS       sequence-317118-        7332 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 7332)
  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 7332)
  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: pCFJ1532 - pEXP[Psmu-1 | mosase (PATCs) | smu-...
FEATURES             Location/Qualifiers
     source          1..7332
                     /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(3928..3952)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(4945..4965)
                     /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(4973..4990)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4973..4989)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4982..5004)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5317..5339
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5377..5395)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5463..5567
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5568..5636,5637..6428)
                     /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(5786..5805)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6599..7187
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7088..7107
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     join(7314..7332,1..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 ggctaaaaaa
     1081 tgtcctcctt cgtcccaaac aaggtaagtt tatctaaaaa gtttttcatt caaaatgtgt
     1141 aaaaattcat ttaaaataac ccaaaaatca ttaatcctcg atattttcag gagcaaaccc
     1201 gtaccgtcct catcttctgc ttccacctca agaagaccgc cgccgagtcc caccgtatgc
     1261 tcgtcgaggt attttcctgc atttttcaac tgggaaaatg aaagaaaatc gataatttca
     1321 ggccttcgga gagcaagtcc caaccgtcaa gacctgcgag cgttggttcc aacgtttcaa
     1381 gtccggagac ttcgacgtcg acgacaagga gcacggaaag ccaccaaagc gttacgagga
     1441 cgccgagctc caagccctcc tcgacgagga cgacgcccaa acgcaaaagc aactcgccga
     1501 gcaactcgag gtatcccaac aagccgtatc caaccgactc cgtgagatgg gaaaggtaag
     1561 ttttcttatg ggaaagaagg aaaaaaccga gattttactt gaaaaattga atttttcgcg
     1621 ggattttcac caaaaattgt tgaatattca ttatttcacg ctgtaaaaca aaaaaaaaaa
     1681 aaatcaaaaa ctacgttgaa atcgcgtttt taagcgaatt ttcttcagaa ttgccagatt
     1741 ttaaccccaa attttgcagt ttttaaataa aatttcacct tttcggctca aattgtagat
     1801 tttcctgaaa atttagtaca aaaaacaatt tcctcgtaaa tttttcaaat agattttcag
     1861 atccaaaagg tcggacgttg ggtcccacac gagctcaacg agcgtcaaat ggagcgtcgt
     1921 aagaacacct gcgagatcct cctctcccgt tacaagcgta agtccttcct ccaccgtatc
     1981 gtcaccggtg acgagaagtg gatcttcttc gtcaacccaa agcgtaaggt aagttatgtc
     2041 tacctgcctg cctaccgcct aaattttttg tgaagtttct tcaaaaaatc cagaaaaaaa
     2101 aacaattttc atacgatttt ttcccttaaa attgtgaatt ttcatgcttt ttagccccaa
     2161 aagtcattat ttgagaaaaa atttcataca aaaaagtttt gagaaataca caatttttta
     2221 aatgtaattt tcaaattttc aattttcaac tagaaaattc acaaaacttg taaattttgg
     2281 accaaaacat ttatacaatt actttttttg aatctaataa ctacaataac tacaaattta
     2341 acattttcag aagtcctacg tcgacccagg acaaccagcc acctccaccg cccgtccaaa
     2401 ccgtttcgga aagaagacca tgctctgcgt ctggtgggac caatccggag tcatctacta
     2461 cgagctcctc aagccaggta agttcttttt ttgaaaagtg cagtttgtag tctaattttc
     2521 atttattttc tttttaaaaa acgcatcaat ttttaaatat tttgggacaa aatccgaaaa
     2581 ctgtactaat ttgtagtttg taaaattaaa aaaaaaacgc aaaaaaatgt tttcaaaaat
     2641 gcttagaata aaaataaaat ttctaaaaaa ttgaacaaaa taaacattta aaaattcaaa
     2701 aagtttgaaa aaatgcaaag ttttttaatg caaaaaatat ttaatgttta aacaaattta
     2761 aaaaatgttg taagaatatg tttagagaca gtttttaaat gttaaattgc aaaaaaaaaa
     2821 agcaaaaaaa aatttgaaaa atgtaacaga aaattttaat taaaaaagtc gtgttttttt
     2881 caaacctgca attctttgta attttattta taaagagaaa caaatcaaat tgtatattac
     2941 gaaaaacgat actaaaattc gaaaatttgc ggttttttgc gtaaaaaata cggttccgta
     3001 attttcagga gaaaccgtca acactgcccg ttaccagcaa cagctcatca atctcaaccg
     3061 tgccctccaa cgtaagcgtc cagagtacca aaagcgtcaa catcgtgtca tcttcctcca
     3121 cgacaacgcc ccatcccaca cggcccgtgc cgtccgtgac accctcgaga cactcaactg
     3181 ggaggtcctc ccacacgccg cctactcccc aggtaagttc atagattttg aaaaaaagtt
     3241 taagaactga aaaatggaat aaaaatattt aagagcattt ttaaatgtaa aattaacaaa
     3301 aaaagcgcct aagaatgttt ccaaaacagt aaaaaaaagg tttaaaaaaa tgcaaaaaaa
     3361 aatttaagat gcttttaatt acacaaaaaa tgaaagtaaa aaaaattaaa atattgaaaa
     3421 atgtatattt gtttgaaaag gtacattttt caatgcaaaa aatgtttcta aaaattaaaa
     3481 aaatatgttt aaattgtgta aagtgtaaat taaaaaataa aattatgaaa aagtattgtt
     3541 tcatactttt tctaaatttt ggttgaaaaa tttaaaattc cagcttttta ttcacattat
     3601 gtgttctaaa aatctttccc caattttcgg gaccgttttc cagtatattt ccatttttca
     3661 aaaagatttt taactgaaat tcatgttttc aatgctaaaa atcaataaaa aaagaatttt
     3721 tcagacctcg ccccatccga ctaccacctc ttcgcctcca tgggacacgc cctcgccgag
     3781 caacgtttcg actcctacga gtccgtcaag aagtggctcg acgagtggtt cgccgccaag
     3841 gacgacgagt tctactggcg tggaatccac aagctcccag agcgttggga gaagtgcgtc
     3901 gcctccgacg gaaagtactt cgagtaaacc cagctttctt gtacaaagtg gattaatttt
     3961 ccccagattt ccccacgatt ttttttgaca tttttatgaa aattttctct aaaagttacc
     4021 cctattttgt ttagactgat ctcttatttt atccaatgtt tcaaattttt gaaattgctc
     4081 gaaaattcca attttcacat atttttgttg attttttcgc cattttcccc tgataaatta
     4141 acttattttt gcaatttttt ctttcgaaaa aatattggtt tctcatcacc atttatcctt
     4201 ttttttatgg agttaaaaat tgttaaaaaa atttaatttt ttttctaatt ttagagtcaa
     4261 aacgggttcg aatctagagc gtagctgtac gattttttgg cgatttttgc ggttttctgg
     4321 tgagtttttt taaagttatt tttttgaaaa aaattcttga aaataaaagg aaattgtttt
     4381 aaaaaaaaat ttcaaagaaa aaattttgaa aaaattgaaa catttttttt attttgacca
     4441 aaaatattga aaattaatta acattttttt ttggaaagtt aatttttaat acaaattttt
     4501 taaatttatt ttacaaaaaa aattgaattt ctgtgaaaaa tcagttttcc agtacataat
     4561 gatttgacca aaaaaaattg aaaaaaatga ttttttaaaa ttaaaaatac cataaaatgc
     4621 tgaaaattca gaaaaataag taaattttaa aaaattctgg aaaaacatct aaaaaccccc
     4681 caaatataat attttggacc cttttatttt ctgctcgaca tctcaaatat cctccaattt
     4741 ttcgcttttt tgattaattg aaagcatagt gggggcaggt gttataggac acccttccct
     4801 ccactttttt ggagcatttt tcaaagctct tccgagagca gaagcttctc aatttttcca
     4861 ccccaaaagt ttgaaattct gccaaaaaat ctcgcatttt aacaattttt cgagatgttt
     4921 cacgtccttt tcattgcgat ttcccaactt tattatacat agttgataat tcactggccg
     4981 tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag
     5041 cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc
     5101 aacagttgcg cagcctgaat ggcgaatggc gcctgatgcg gtattttctc cttacgcatc
     5161 tgtgcggtat ttcacaccgc atatggtgca ctctcagtgc aatctgctct gatgccgcat
     5221 agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc
     5281 tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt
     5341 tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat
     5401 aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg
     5461 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     5521 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     5581 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     5641 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     5701 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     5761 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     5821 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     5881 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     5941 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     6001 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     6061 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     6121 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     6181 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     6241 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     6301 cagcactggg gtcagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     6361 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     6421 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     6481 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     6541 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     6601 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     6661 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     6721 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     6781 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     6841 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     6901 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     6961 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     7021 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     7081 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     7141 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     7201 cggccttttt acggttcctg gtcttttgct ggccttttgc tcacatgttc tttcctgcgt
     7261 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accagcggat
     7321 aacaatttca ca
//
