LOCUS       sequence-317361-       10282 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 10282)
  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 10282)
  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: pMDJ23 - pEXP[Psmu-2 | eFlp recombinase (PATCs...
FEATURES             Location/Qualifiers
     source          1..10282
                     /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    2121..2145
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(4633..4657)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(7895..7915)
                     /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(7923..7940)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7923..7939)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7932..7954)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8267..8289
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(8327..8345)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        8413..8517
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8518..8586,8587..9378)
                     /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(8736..8755)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9549..10137
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10038..10057
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     join(10264..10282,1..4)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
ORIGIN
        1 caggaaacag ctatgaccat gattacgcca agctatcaac tttgtataga aaagttgtgc
       61 accaatttga gtttccatgg ggaatcaagc ggataccgat tgagttgaag aaccctaaat
      121 aaaattttat atattttacc ttccaacttc gaaaaaatca ctcaccattt tagcatgttt
      181 ccctcaaaaa ccgtggcaaa ttgctcttgg aaaataatag acacatcagt tggtcgagca
      241 ttttttggag gaattttaaa actgtgatga tggttgaagg tgactatatt gtctgcgagt
      301 tgatactctg gaatattgat ttttttgaag atttcatagt cttcttggat ttcaaaaatt
      361 gaaatatttc agctgtatta aaagagaata aataaaatga tattgcctag gggtgagcgg
      421 caaacgattt tttccggcaa atcggcaaat cgacaaattg ccggaattgg aaatttccag
      481 caaatcggca tatcggcaat ttgccggaat ttaaaatttc cggcaaaccg gcaaaccggc
      541 aaattgccga tttgccgaac ttcggtaaat ttgcaaatcg gtaatttgcc gaaaattttc
      601 ggcaaattgt ggttttgcac ttattttttg gaatttcaat ttcaaatggc aaaattaaac
      661 gcatcatatg aatttttcta tatctatttt gagaagttct atgaaaatat gtagaaaaaa
      721 cgggaaaaat tcaataatag acagttttaa gtgcttccgt cttataaaaa atccctctaa
      781 acatttccgg caaatctggt attcggcaaa tcggcaagtc ggcaatttgc cgtaaattaa
      841 aatttccggc aaaacggcaa accggcaaat tgccggaatt ggaaatttcc ggcaaatcgg
      901 caaattgctt gcccattttt tgattcttag tcggcgtttt acacaaaaga taaattccaa
      961 taaaagacag aactttgctt tttgccagaa attaaaaaat tggggcaaac caatccaaat
     1021 atgaagtgta aacttgggtt tgcaccaggg ctgtgcggaa accggcaatt gccgaaattg
     1081 ccgaacccta aaaactttgg caaccggcaa cttttgctgg ctggaacttc ggcaactttt
     1141 cggcaaacgg caacttggtt gccggttgcg caaagctctg gtttgcgttt agtagcaagg
     1201 cttcctgaaa gaattccgga ggaaaactcc ccctaactca ccttttactc tatgcatctg
     1261 cttaattttg tgcaattgta gtgcaaatag taggaaatga attttttttc cgccattttt
     1321 tccttttcct atttctgctt ttcaatttcc acgtggagct cttttaaaat gaaaatttaa
     1381 actctagata tttcacgttt taaaaaaaaa aaattttaat tttctatcgg ttttactttt
     1441 gacgaaaagg gttcacagac attggaaata atcatcattt actatatata aagcgcttat
     1501 tctgtgtgtc catagtttgt agtctctgta gtctttgtag tctgtgaagt tttggcttct
     1561 ggagggatag tgagttgggg tcagtgtagg gatatagtcg gggtactgta gtggtacaat
     1621 agtggtacgg taggaatact gtagggttac ggtagtttca taaaaaatta attttcagcc
     1681 ccagaagtcg ggggccgcgc cgaaggtgcc tagacctaag cctaggcctc agcctaagcc
     1741 taattacatc aaaaaagcta tgacttttgg atcattttga cttcttttct agaacgcata
     1801 atgaaataat tcttaatttc atcggctttt tctcatcgtt gcttagaagt attgtcttga
     1861 aaattgatat ttttagaaag tacacatatt tctcaatcct tttatttaaa tttctgaaag
     1921 aaactttgcc acaacggtga gaaaagggtg cccgaaaaat gtgcatttaa atgtgaaaaa
     1981 tatccgtggc cgcggcctaa gaacccccgt cggccatgtg gtgcaagtgc gctctgttgc
     2041 ggatttttat atttaattgc attttcgcat attcgaacga ttttcctgat tttccagccg
     2101 aaattgtcca ttaaaaagtg gcaagtttgt acaaaaaagc aggctaaaaa tgtcccaatt
     2161 cgacatcctc tgcaaggtat tttcctgcat ttttcaactg ggaaaatgaa agaaaatcga
     2221 taatttcaga ccccaccaaa ggtcctcgtc cgtcaattcg tcgagcgttt cgagcgtcca
     2281 tccggagaga agatcgcctc ctgcgccgcc gagctcacct acctctgctg gatgatcacc
     2341 cacaacggaa ccgccatcaa gcgtgccacc ttcatgtcct acaacaccat catctccaac
     2401 tccctctcct tcgacatcgt caacaagtcc ctccaattca agtacaaggt aagttttctt
     2461 atgggaaaga aggaaaaaac cgagatttta cttgaaaaat tgaatttttc gcgggatttt
     2521 caccaaaaat tgttgaatat tcattatttc acgctgtaaa acaaaaaaaa aaaaaatcaa
     2581 aaactacgtt gaaatcgcgt ttttaagcga attttcttca gaattgccag attttaaccc
     2641 caaattttgc agtttttaaa taaaatttca ccttttcggc tcaaattgta gattttcctg
     2701 aaaatttagt acaaaaaaca atttcctcgt aaatttttca aatagatttt cagacgcaaa
     2761 aggccaccat cctcgaggcc tccctcaaga agctcatccc agcctgggag ttcaccatca
     2821 tcccatacaa cggacaaaag caccaatccg acatcaccga catcgtctcc tccctccaac
     2881 tccaattcga gtcctccgag gaggccgaca agggaaactc ccactccaag aagatgctca
     2941 aggccctcct ctccgaggga gagtccatct gggagatcac cgagaagatc ctcaactcct
     3001 tcgagtacac ctcccgtttc accaagacca agaccctcta ccaattcctc ttcctcgcca
     3061 ccttcatcaa ctgcggacgt ttctccgaca tcaaggtaag ttatgtctac ctgcctgcct
     3121 accgcctaaa ttttttgtga agtttcttca aaaaatccag aaaaaaaaac aattttcata
     3181 cgattttttc ccttaaaatt gtgaattttc atgcttttta gccccaaaag tcattatttg
     3241 agaaaaaatt tcatacaaaa aagttttgag aaatacacaa ttttttaaat gtaattttca
     3301 aattttcaat tttcaactag aaaattcaca aaacttgtaa attttggacc aaaacattta
     3361 tacaattact ttttttgaat ctaataacta caataactac aaatttaaca ttttcagaac
     3421 gtcgacccaa agtccttcaa gctcgtccaa aacaagtacc tcggagtcat catccaatgc
     3481 ctcgtcaccg agacgaagac ctccgtctcc cgtcacatct acttcttctc cgcccgtgga
     3541 cgtatcgacc cactcgtcta cctcgacgag ttcctccgta actccgagcc agtcctcaag
     3601 cgtgtcaacc gtaccggaaa ctcctcctcc aacaagcaag agtaccaact cctcaaggac
     3661 aacctcgtcc gttcctacaa caaggccctc aagaagaacg ccccataccc aatcttcgcc
     3721 atcaaggtaa gttctttttt tgaaaagtgc agtttgtagt ctaattttca tttattttct
     3781 ttttaaaaaa cgcatcaatt tttaaatatt ttgggacaaa atccgaaaac tgtactaatt
     3841 tgtagtttgt aaaattaaaa aaaaaacgca aaaaaatgtt ttcaaaaatg cttagaataa
     3901 aaataaaatt tctaaaaaat tgaacaaaat aaacatttaa aaattcaaaa agtttgaaaa
     3961 aatgcaaagt tttttaatgc aaaaaatatt taatgtttaa acaaatttaa aaaatgttgt
     4021 aagaatatgt ttagagacag tttttaaatg ttaaattgca aaaaaaaaaa gcaaaaaaaa
     4081 atttgaaaaa tgtaacagaa aattttaatt aaaaaagtcg tgtttttttc aaacctgcaa
     4141 ttctttgtaa ttttatttat aaagagaaac aaatcaaatt gtatattacg aaaaacgata
     4201 ctaaaattcg aaaatttgcg gttttttgcg taaaaaatac ggttccgtaa ttttcagaac
     4261 ggaccaaagt cccacatcgg acgtcacctc atgacctcct tcctctccat gaagggactc
     4321 accgagctca ccaacgtcgt cggaaactgg tccgacaagc gtgcctccgc cgtcgcccgt
     4381 accacctaca cccaccaaat caccgccatc ccagaccact acttcgccct cgtctcccgt
     4441 tactacgcct acgacccaat ctccaaggag atgatcgccc tcaaggacga gacaaaccca
     4501 atcgaggagt ggcaacacat cgagcaactc aagggatccg ccgagggatc catccgttac
     4561 ccagcctgga acggaatcat ctcccaagag gtcctcgact acctctcctc ctacatcaac
     4621 cgtcgtatct agacccagct ttcttgtaca aagtggccta agctgtctca tcctactttc
     4681 acctagttaa ctgcttgtct taaaatctat gcttctcttt agtatctaaa attttcctag
     4741 aagcttacaa gtatataaat ggtctcttct caataaaggt tgtatattta ttcatcttat
     4801 tgaatctgcc atttcctcgt ttttgcgagt ttatatacct tccaattttc tttctattgt
     4861 attttcaact tctaatttta attcagggaa actgcttcaa cgcatcatgg gttgtaccat
     4921 gtcacaggaa gagcgtgccg ctcttgaaag atcacgaatg tcctcagtct ccaaggtaac
     4981 acttagtttt tgttgctctg tttaaaaatt aattttaagg gagaggagct catcaaggag
     5041 aacatgcaca tgaagctcta catggaggga accgtcaaca accaccactt caagtgcacc
     5101 tccgagggag agggaaagcc atacgaggga acccaaacca tgcgtatcaa ggtcgtcgag
     5161 ggaggaccac tcccattcgc cttcgacatc ctcgccacct ccttcatgta cggatcgcgt
     5221 accttcatca accacaccca aggtaagttt tcttatggga aagaaggaaa aaaccgagat
     5281 tttacttgaa aaattgaatt tttcgcggga ttttcaccaa aaattgttga atattcatta
     5341 tttcacgctg taaaacaaaa aaaaaaaaaa tcaaaaacta cgttgaaatc gcgtttttaa
     5401 gcgaattttc ttcagaattg ccagatttta accccaaatt ttgcagtttt taaataaaat
     5461 ttcacctttt cggctcaaat tgtagatttt cctgaaaatt tagtacaaaa aacaatttcc
     5521 tcgtaaattt ttcaaataga ttttcaggaa tcccagactt cttcaagcaa tccttcccag
     5581 agggattcac ctgggagcgt gtcaccacct acgaggacgg aggagtcctc accgccaccc
     5641 aagacacctc cctccaagac ggatgcctca tctacaacgt caaggtaagt tatgtctacc
     5701 tgcctgccta ccgcctaaat tttttgtgaa gtttcttcaa aaaatccaga aaaaaaaaca
     5761 attttcatac gattttttcc cttaaaattg tgaattttca tgctttttag ccccaaaagt
     5821 cattatttga gaaaaaattt catacaaaaa agttttgaga aatacacaat tttttaaatg
     5881 taattttcaa attttcaatt ttcaactaga aaattcacaa aacttgtaaa ttttggacca
     5941 aaacatttat acaattactt tttttgaatc taataactac aataactaca aatttaacat
     6001 tttcagatcc gtggagtcaa cttcccatcc aacggaccag tcatgcaaaa gaagaccctc
     6061 ggatgggagg ccaacaccga gatgctctac ccagccgacg gaggactgga gggacgtacc
     6121 gacatggccc tcaagctcgt cggaggagga cacctcatct gcaacttcaa gaccacctac
     6181 cgttccaaga agccagccaa ggtaagttct ttttttgaaa agtgcagttt gtagtctaat
     6241 tttcatttat tttcttttta aaaaacgcat caatttttaa atattttggg acaaaatccg
     6301 aaaactgtac taatttgtag tttgtaaaat taaaaaaaaa acgcaaaaaa atgttttcaa
     6361 aaatgcttag aataaaaata aaatttctaa aaaattgaac aaaataaaca tttaaaaatt
     6421 caaaaagttt gaaaaaatgc aaagtttttt aatgcaaaaa atatttaatg tttaaacaaa
     6481 tttaaaaaat gttgtaagaa tatgtttaga gacagttttt aaatgttaaa ttgcaaaaaa
     6541 aaaaagcaaa aaaaaatttg aaaaatgtaa cagaaaattt taattaaaaa agtcgtgttt
     6601 ttttcaaacc tgcaattctt tgtaatttta tttataaaga gaaacaaatc aaattgtata
     6661 ttacgaaaaa cgatactaaa attcgaaaat ttgcggtttt ttgcgtaaaa aatacggttc
     6721 cgtaattttc agaacctcaa gatgccagga gtctactacg tcgaccaccg tctggagcgt
     6781 atcaaggagg ccgacaagga gacgtacgtc gagcaacacg aggtcgccgt cgcccgttac
     6841 tgcgacctcc catccaagct cggacacaag ctcaacggaa tggacgagct ctacaagtcg
     6901 ggataatttt ccccagattt ccccacgatt ttttttgaca tttttatgaa aattttctct
     6961 aaaagttacc cctattttgt ttagactgat ctcttatttt atccaatgtt tcaaattttt
     7021 gaaattgctc gaaaattcca attttcacat atttttgttg attttttcgc cattttcccc
     7081 tgataaatta acttattttt gcaatttttt ctttcgaaaa aatattggtt tctcatcacc
     7141 atttatcctt ttttttatgg agttaaaaat tgttaaaaaa atttaatttt ttttctaatt
     7201 ttagagtcaa aacgggttcg aatctagagc gtagctgtac gattttttgg cgatttttgc
     7261 ggttttctgg tgagtttttt taaagttatt tttttgaaaa aaattcttga aaataaaagg
     7321 aaattgtttt aaaaaaaaat ttcaaagaaa aaattttgaa aaaattgaaa catttttttt
     7381 attttgacca aaaatattga aaattaatta acattttttt ttggaaagtt aatttttaat
     7441 acaaattttt taaatttatt ttacaaaaaa aattgaattt ctgtgaaaaa tcagttttcc
     7501 agtacataat gatttgacca aaaaaaattg aaaaaaatga ttttttaaaa ttaaaaatac
     7561 cataaaatgc tgaaaattca gaaaaataag taaattttaa aaaattctgg aaaaacatct
     7621 aaaaaccccc caaatataat attttggacc cttttatttt ctgctcgaca tctcaaatat
     7681 cctccaattt ttcgcttttt tgattaattg aaagcatagt gggggcaggt gttataggac
     7741 acccttccct ccactttttt ggagcatttt tcaaagctct tccgagagca gaagcttctc
     7801 aatttttcca ccccaaaagt ttgaaattct gccaaaaaat ctcgcatttt aacaattttt
     7861 cgagatgttt cacgtccttt tcattgcgat ttcccaactt tattatacat agttgataat
     7921 tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat
     7981 cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat
     8041 cgcccttccc aacagttgcg cagcctgaat ggcgaatggc gcctgatgcg gtattttctc
     8101 cttacgcatc tgtgcggtat ttcacaccgc atatggtgca ctctcagtgc aatctgctct
     8161 gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg
     8221 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg
     8281 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc
     8341 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt
     8401 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     8461 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     8521 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     8581 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     8641 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     8701 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     8761 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     8821 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     8881 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     8941 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     9001 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     9061 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     9121 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     9181 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tggctctcgc
     9241 ggtatcattg cagcactggg gtcagatggt aagccctccc gtatcgtagt tatctacacg
     9301 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     9361 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     9421 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     9481 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     9541 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     9601 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     9661 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     9721 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     9781 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     9841 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     9901 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     9961 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
    10021 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
    10081 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
    10141 gccagcaacg cggccttttt acggttcctg gtcttttgct ggccttttgc tcacatgttc
    10201 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
    10261 accagcggat aacaatttca ca
//
