LOCUS       sequence-317354-       10042 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 10042)
  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 10042)
  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: pMDJ22 - pEXP[Psmu-2 | Cre Recombinase (PATCs)...
FEATURES             Location/Qualifiers
     source          1..10042
                     /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(4393..4417)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(7655..7675)
                     /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(7683..7700)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7683..7699)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7692..7714)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8027..8049
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(8087..8105)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        8173..8277
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8278..8346,8347..9138)
                     /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(8496..8515)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9309..9897
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9798..9817
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     join(10024..10042,1..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 tgtccaacct
     2161 cctcaccgtc caccaaaacc tcccagccct cccagtcgac gccacctccg acgaggtccg
     2221 taaggtattt tcctgcattt ttcaactggg aaaatgaaag aaaatcgata atttcagaac
     2281 ctcatggaca tgttccgtga ccgtcaagcc ttctccgagc acacctggaa gatgctcctc
     2341 tccgtctgcc gttcctgggc cgcctggtgc aagctcaaca accgtaagtg gttcccagcc
     2401 gagccaggta agttttctta tgggaaagaa ggaaaaaacc gagattttac ttgaaaaatt
     2461 gaatttttcg cgggattttc accaaaaatt gttgaatatt cattatttca cgctgtaaaa
     2521 caaaaaaaaa aaaaatcaaa aactacgttg aaatcgcgtt tttaagcgaa ttttcttcag
     2581 aattgccaga ttttaacccc aaattttgca gtttttaaat aaaatttcac cttttcggct
     2641 caaattgtag attttcctga aaatttagta caaaaaacaa tttcctcgta aatttttcaa
     2701 atagattttc agaggacgtc cgtgactacc tcctctacct ccaagcccgt ggactcgccg
     2761 tcaagaccat ccaacaacac ctcggacaac tcaacatgct ccaccgtcgt tccggactcc
     2821 cacgtccatc cgactccaac gccgtctccc tcgtcatgcg tcgtatccgt aaggagaacg
     2881 tcgacgccgg agagcgtgcc aagcaagccc tcgccttcga gcgtaccgac ttcgaccaag
     2941 tccgttccct catggagaac tccgaccgtt gccaaggtaa gttatgtcta cctgcctgcc
     3001 taccgcctaa attttttgtg aagtttcttc aaaaaatcca gaaaaaaaaa caattttcat
     3061 acgatttttt cccttaaaat tgtgaatttt catgcttttt agccccaaaa gtcattattt
     3121 gagaaaaaat ttcatacaaa aaagttttga gaaatacaca attttttaaa tgtaattttc
     3181 aaattttcaa ttttcaacta gaaaattcac aaaacttgta aattttggac caaaacattt
     3241 atacaattac tttttttgaa tctaataact acaataacta caaatttaac attttcagac
     3301 atccgtaacc tcgccttcct cggaatcgcc tacaacaccc tcctccgtat cgccgagatc
     3361 gcccgtatcc gtgtcaagga catctcccgt accgacggag gacgtatgct catccacatc
     3421 ggacgtacca agaccctcgt ctccaccgcc ggagtcgaga aggccctctc cctcggagtc
     3481 accaagctcg tcgagcgttg gatctccgtc tccggagtcg ccgacgaccc aaacaactac
     3541 ctcttctgcc gtgtccgtaa ggtaagttct ttttttgaaa agtgcagttt gtagtctaat
     3601 tttcatttat tttcttttta aaaaacgcat caatttttaa atattttggg acaaaatccg
     3661 aaaactgtac taatttgtag tttgtaaaat taaaaaaaaa acgcaaaaaa atgttttcaa
     3721 aaatgcttag aataaaaata aaatttctaa aaaattgaac aaaataaaca tttaaaaatt
     3781 caaaaagttt gaaaaaatgc aaagtttttt aatgcaaaaa atatttaatg tttaaacaaa
     3841 tttaaaaaat gttgtaagaa tatgtttaga gacagttttt aaatgttaaa ttgcaaaaaa
     3901 aaaaagcaaa aaaaaatttg aaaaatgtaa cagaaaattt taattaaaaa agtcgtgttt
     3961 ttttcaaacc tgcaattctt tgtaatttta tttataaaga gaaacaaatc aaattgtata
     4021 ttacgaaaaa cgatactaaa attcgaaaat ttgcggtttt ttgcgtaaaa aatacggttc
     4081 cgtaattttc agaacggagt cgccgcccca tccgccacct cccaactctc cacccgtgcc
     4141 ctcgagggaa tcttcgaggc cacccaccgt ctcatctacg gagccaagga cgactccgga
     4201 caacgttacc tcgcctggtc cggacactcc gcccgtgtcg gagccgcccg tgacatggcc
     4261 cgtgccggag tctccatccc agagatcatg caagccggag gatggaccaa cgtcaacatc
     4321 gtcatgaact acatccgtaa cctcgactcc gagacgggag ccatggtccg tctcctcgag
     4381 gacggagact agacccagct ttcttgtaca aagtggccta agctgtctca tcctactttc
     4441 acctagttaa ctgcttgtct taaaatctat gcttctcttt agtatctaaa attttcctag
     4501 aagcttacaa gtatataaat ggtctcttct caataaaggt tgtatattta ttcatcttat
     4561 tgaatctgcc atttcctcgt ttttgcgagt ttatatacct tccaattttc tttctattgt
     4621 attttcaact tctaatttta attcagggaa actgcttcaa cgcatcatgg gttgtaccat
     4681 gtcacaggaa gagcgtgccg ctcttgaaag atcacgaatg tcctcagtct ccaaggtaac
     4741 acttagtttt tgttgctctg tttaaaaatt aattttaagg gagaggagct catcaaggag
     4801 aacatgcaca tgaagctcta catggaggga accgtcaaca accaccactt caagtgcacc
     4861 tccgagggag agggaaagcc atacgaggga acccaaacca tgcgtatcaa ggtcgtcgag
     4921 ggaggaccac tcccattcgc cttcgacatc ctcgccacct ccttcatgta cggatcgcgt
     4981 accttcatca accacaccca aggtaagttt tcttatggga aagaaggaaa aaaccgagat
     5041 tttacttgaa aaattgaatt tttcgcggga ttttcaccaa aaattgttga atattcatta
     5101 tttcacgctg taaaacaaaa aaaaaaaaaa tcaaaaacta cgttgaaatc gcgtttttaa
     5161 gcgaattttc ttcagaattg ccagatttta accccaaatt ttgcagtttt taaataaaat
     5221 ttcacctttt cggctcaaat tgtagatttt cctgaaaatt tagtacaaaa aacaatttcc
     5281 tcgtaaattt ttcaaataga ttttcaggaa tcccagactt cttcaagcaa tccttcccag
     5341 agggattcac ctgggagcgt gtcaccacct acgaggacgg aggagtcctc accgccaccc
     5401 aagacacctc cctccaagac ggatgcctca tctacaacgt caaggtaagt tatgtctacc
     5461 tgcctgccta ccgcctaaat tttttgtgaa gtttcttcaa aaaatccaga aaaaaaaaca
     5521 attttcatac gattttttcc cttaaaattg tgaattttca tgctttttag ccccaaaagt
     5581 cattatttga gaaaaaattt catacaaaaa agttttgaga aatacacaat tttttaaatg
     5641 taattttcaa attttcaatt ttcaactaga aaattcacaa aacttgtaaa ttttggacca
     5701 aaacatttat acaattactt tttttgaatc taataactac aataactaca aatttaacat
     5761 tttcagatcc gtggagtcaa cttcccatcc aacggaccag tcatgcaaaa gaagaccctc
     5821 ggatgggagg ccaacaccga gatgctctac ccagccgacg gaggactgga gggacgtacc
     5881 gacatggccc tcaagctcgt cggaggagga cacctcatct gcaacttcaa gaccacctac
     5941 cgttccaaga agccagccaa ggtaagttct ttttttgaaa agtgcagttt gtagtctaat
     6001 tttcatttat tttcttttta aaaaacgcat caatttttaa atattttggg acaaaatccg
     6061 aaaactgtac taatttgtag tttgtaaaat taaaaaaaaa acgcaaaaaa atgttttcaa
     6121 aaatgcttag aataaaaata aaatttctaa aaaattgaac aaaataaaca tttaaaaatt
     6181 caaaaagttt gaaaaaatgc aaagtttttt aatgcaaaaa atatttaatg tttaaacaaa
     6241 tttaaaaaat gttgtaagaa tatgtttaga gacagttttt aaatgttaaa ttgcaaaaaa
     6301 aaaaagcaaa aaaaaatttg aaaaatgtaa cagaaaattt taattaaaaa agtcgtgttt
     6361 ttttcaaacc tgcaattctt tgtaatttta tttataaaga gaaacaaatc aaattgtata
     6421 ttacgaaaaa cgatactaaa attcgaaaat ttgcggtttt ttgcgtaaaa aatacggttc
     6481 cgtaattttc agaacctcaa gatgccagga gtctactacg tcgaccaccg tctggagcgt
     6541 atcaaggagg ccgacaagga gacgtacgtc gagcaacacg aggtcgccgt cgcccgttac
     6601 tgcgacctcc catccaagct cggacacaag ctcaacggaa tggacgagct ctacaagtcg
     6661 ggataatttt ccccagattt ccccacgatt ttttttgaca tttttatgaa aattttctct
     6721 aaaagttacc cctattttgt ttagactgat ctcttatttt atccaatgtt tcaaattttt
     6781 gaaattgctc gaaaattcca attttcacat atttttgttg attttttcgc cattttcccc
     6841 tgataaatta acttattttt gcaatttttt ctttcgaaaa aatattggtt tctcatcacc
     6901 atttatcctt ttttttatgg agttaaaaat tgttaaaaaa atttaatttt ttttctaatt
     6961 ttagagtcaa aacgggttcg aatctagagc gtagctgtac gattttttgg cgatttttgc
     7021 ggttttctgg tgagtttttt taaagttatt tttttgaaaa aaattcttga aaataaaagg
     7081 aaattgtttt aaaaaaaaat ttcaaagaaa aaattttgaa aaaattgaaa catttttttt
     7141 attttgacca aaaatattga aaattaatta acattttttt ttggaaagtt aatttttaat
     7201 acaaattttt taaatttatt ttacaaaaaa aattgaattt ctgtgaaaaa tcagttttcc
     7261 agtacataat gatttgacca aaaaaaattg aaaaaaatga ttttttaaaa ttaaaaatac
     7321 cataaaatgc tgaaaattca gaaaaataag taaattttaa aaaattctgg aaaaacatct
     7381 aaaaaccccc caaatataat attttggacc cttttatttt ctgctcgaca tctcaaatat
     7441 cctccaattt ttcgcttttt tgattaattg aaagcatagt gggggcaggt gttataggac
     7501 acccttccct ccactttttt ggagcatttt tcaaagctct tccgagagca gaagcttctc
     7561 aatttttcca ccccaaaagt ttgaaattct gccaaaaaat ctcgcatttt aacaattttt
     7621 cgagatgttt cacgtccttt tcattgcgat ttcccaactt tattatacat agttgataat
     7681 tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat
     7741 cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat
     7801 cgcccttccc aacagttgcg cagcctgaat ggcgaatggc gcctgatgcg gtattttctc
     7861 cttacgcatc tgtgcggtat ttcacaccgc atatggtgca ctctcagtgc aatctgctct
     7921 gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg
     7981 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg
     8041 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc
     8101 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt
     8161 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     8221 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     8281 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     8341 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     8401 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     8461 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     8521 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     8581 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     8641 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     8701 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     8761 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     8821 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     8881 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     8941 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tggctctcgc
     9001 ggtatcattg cagcactggg gtcagatggt aagccctccc gtatcgtagt tatctacacg
     9061 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     9121 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     9181 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     9241 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     9301 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     9361 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     9421 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     9481 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     9541 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     9601 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     9661 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     9721 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     9781 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     9841 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     9901 gccagcaacg cggccttttt acggttcctg gtcttttgct ggccttttgc tcacatgttc
     9961 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
    10021 accagcggat aacaatttca ca
//
