LOCUS       sequence-376954-       10508 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Heat shock inducible Cas9 expression in C. elegans..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10508)
  AUTHORS   El Mouridi S, Alkhaldi F, Frokjaer-Jensen C
  TITLE     Modular safe-harbor transgene insertion (MosTI) for targeted 
            single-copy and extrachromosomal array integration in C. elegans.
  JOURNAL   G3 (Bethesda). 2022 Jul 28. pii: 6651068. doi: 
            10.1093/g3journal/jkac184.
  PUBMED    35900171
REFERENCE   2  (bases 1 to 10508)
  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: "G3 
            (Bethesda). 2022 Jul 28. pii: 6651068. doi: 
            10.1093/g3journal/jkac184."
COMMENT     Sequence Label: pMDJ231 - Phsp Cas9
FEATURES             Location/Qualifiers
     source          1..10508
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     3..25
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     22..38
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     22..38
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    58..78
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     protein_bind    424..448
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             4829..4849
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4850..4876
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     protein_bind    complement(4880..4904)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(8142..8162)
                     /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(8170..8187)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(8170..8186)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8179..8201)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8514..8536
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(8574..8592)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        8660..8764
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8765..8833,8834..9625)
                     /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(8983..9002)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9796..10384
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10285..10304
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccagcggata acaatttcac acaggaaaca gctatgacca tgattacgcc aagctatcaa
       61 ctttgtatag aaaagttgga ttatagtttg aagatttcta atttcacaat tagagcaaat
      121 gttgttcggt atttattttc aacggtattt atactatttt ccaccttttt ctagaacatt
      181 cgagctgctt gttgcaaaag gagggcgact cacattcggt acatggaaaa gtagtgtaca
      241 caataaagag acccagatac attttccgtc tgcgtctctt tgcacccacc gggagtattt
      301 tcaaacgaat gcatctagga ccttctagaa cattctgtaa ggctgcagaa tgcgggtata
      361 taaggaaagc gggctcagag gaagccaaca cgctttgttc tagtgcatct aaaaaacttc
      421 gaaacaagtt tgtacaaaaa agcaggctaa aaatggacaa aaaatacagc atcggccttg
      481 acatcggaac gaactccgtc ggatgggccg tcatcaccga cgagtacaag gtattttcct
      541 gcatttttca actgggaaaa tgaaagaaaa tcgataattt caggtcccat ccaagaagtt
      601 caaggtcctc ggaaacaccg accgtcactc catcaagaag aacctcatcg gagccctcct
      661 cttcgactcc ggagagactg ccgaggccac ccgtctcaag cgtaccgccc gtcgtcgtta
      721 cacccgtcgt aagaaccgta tctgctacct ccaagagatc ttctccaacg agatggccaa
      781 ggtcgacgac tccttcttcc accgtctcga ggagtccttc ctcgtcgagg aggacaagaa
      841 gcacgagcgt cacccaatct tcggaaacat cgtcgacgag gtcgcctacc acgagaagta
      901 cccaaccatc taccacctcc gtaagaagct cgtcgactcc accgacaagg ccgacctccg
      961 tctcatctac ctcgccctcg cccacatgat caagttccgt ggacacttcc tcatcgaggg
     1021 tgacctcaac ccagacaact ccgacgtcga caagctcttc atccaactcg tccaaaccta
     1081 caaccaactc ttcgaggaga acccaatcaa cgcctccgga gtcgacgcca aggccatcct
     1141 ctccgcccgt ctctccaagt cccgtcgtct cgagaacctc atcgcccaac tcccaggaga
     1201 gaagaagaac ggactcttcg gaaacctcat cgccctctcc ctcggactca ccccaaactt
     1261 caagtccaac ttcgacctcg ccgaggacgc caagctccaa ctctccaagg acacctacga
     1321 cgacgacctc gacaacctcc tcgcccaaat cggtgaccaa tacgccgacc tcttcctcgc
     1381 cgccaagaac ctctccgacg ccatcctcct ctccgacatc ctccgtgtca acaccgagat
     1441 caccaaggcc ccactctccg cctccatgat caagcgttac gacgagcacc accaagacct
     1501 caccctcctc aaggccctcg tccgtcaaca actcccagag aagtacaagg agatcttctt
     1561 cgaccaatcc aaggtaagtt ttctagagac ctttactaac taactaggtc tcatagattt
     1621 tcagaacgga tacgccggat acatcgacgg aggagcctcc caagaggagt tctacaagtt
     1681 catcaagcca atcctcgaga agatggacgg aaccgaggag ctcctcgtca agctcaaccg
     1741 tgaggacctc ctccgtaagc aacgtacctt cgacaacgga tccatcccac accaaatcca
     1801 cctcggagag ctccacgcca tcctccgtcg tcaagaggac ttctacccat tcctcaagga
     1861 caaccgtgag aagatcgaga agatcctcac cttccgtatc ccatactacg tcggaccact
     1921 cgcccgtgga aactcccgtt tcgcctggat gacccgtaag tccgaggaga ctatcacccc
     1981 atggaacttc gaggaggtcg tcgacaaggg agcctccgcc caatccttca tcgagcgtat
     2041 gaccaacttc gacaagaacc tcccaaacga gaaggtcctc ccaaagcact ccctcctcta
     2101 cgagtacttc accgtctaca acgagctcac caaggtcaag tacgtcaccg agggaatgcg
     2161 taagccagcc ttcctctccg gagagcaaaa gaaggccatc gtcgacctcc tcttcaagac
     2221 caaccgtaag gtcaccgtca agcaactcaa ggaggactac ttcaagaaga tcgagtgctt
     2281 cgactccgtc gagatctccg gagtcgagga ccgtttcaac gcctccctcg gaacctacca
     2341 cgacctcctc aagatcatca aggacaagga cttcctcgac aacgaggaga acgaggacat
     2401 cctcgaggac atcgtcctca ccctcaccct cttcgaggac cgtgagatga tcgaggagcg
     2461 tctcaagacc tacgcccacc tcttcgacga caaggtcatg aagcaactca agcgtcgtcg
     2521 ttacaccgga tggggacgtc tctcccgtaa gctcatcaac ggaatccgtg acaagcaatc
     2581 cggaaagacc atcctcgact tcctcaagtc cgacggattc gccaaccgta acttcatgca
     2641 actcatccac gacgactccc tcaccttcaa ggtaagttat gtagagacct ttactaacta
     2701 actaggtctc aaacattttc aggaggacat ccaaaaggcc caagtctccg gacaaggaga
     2761 ctccctccac gagcacatcg ccaacctcgc cggatcccca gccatcaaga agggaatcct
     2821 ccaaaccgtc aaggtcgtcg acgagctcgt caaggtcatg ggacgtcaca agccagagaa
     2881 catcgtcatc gagatggccc gtgagaacca aaccacccaa aagggacaaa agaactcccg
     2941 tgagcgtatg aagcgtatcg aggagggaat caaggagctc ggatcccaaa tcctcaagga
     3001 gcacccagtc gagaacaccc aactccaaaa cgagaagctc tacctctact acctccaaaa
     3061 cggacgtgac atgtacgtcg accaagagct cgacatcaac cgtctctccg actacgacgt
     3121 cgaccacatc gtcccacaat ccttcctcaa ggacgactcc atcgacaaca aggtcctcac
     3181 ccgttccgac aagaaccgtg gaaagtccga caacgtccca tccgaggagg tcgtcaagaa
     3241 gatgaagaac tactggcgtc aactcctcaa cgccaagctc atcacccaac gtaagttcga
     3301 caacctcacc aaggccgagc gtggaggact ctccgagctc gacaaggccg gattcatcaa
     3361 gcgtcaactc gtcgagactc gtcaaatcac caagcacgtc gcccaaatcc tcgactcccg
     3421 tatgaacacc aagtacgacg agaacgacaa gctcatccgt gaggtcaagg tcatcaccct
     3481 caagtccaag ctcgtctccg acttccgtaa ggacttccaa ttctacaagg tccgtgagat
     3541 caacaactac caccacgccc acgacgccta cctcaacgcc gtcgtcggaa ccgccctcat
     3601 caagaagtac ccaaagctcg agtccgagtt cgtctacgga gactacaagg tctacgacgt
     3661 ccgtaagatg atcgccaagt ccgagcaaga gatcggaaag gtaagttctt tagagacctt
     3721 tactaactaa ctaggtctca gtaattttca ggccaccgcc aagtacttct tctactccaa
     3781 catcatgaac ttcttcaaga ccgagatcac cctcgccaac ggagagatcc gtaagcgtcc
     3841 actcatcgag actaacggag agactggaga gatcgtctgg gacaagggac gtgacttcgc
     3901 caccgtccgt aaggtcctct ccatgccaca agtcaacatc gtcaagaaga ccgaggtcca
     3961 aaccggagga ttctccaagg agtccatcct cccaaagcgt aactccgaca agctcatcgc
     4021 ccgtaagaag gactgggacc caaagaagta cggaggattc gactccccaa ccgtcgccta
     4081 ctccgtcctc gtcgtcgcca aggtcgagaa gggaaagtcc aagaagctca agtccgtcaa
     4141 ggagctcctc ggaatcacca tcatggagcg ttcctccttc gagaagaacc caatcgactt
     4201 cctcgaggcc aagggataca aggaggtcaa gaaggacctc atcatcaagc tcccaaagta
     4261 ctccctcttc gagctcgaga acggacgtaa gcgtatgctc gcctccgccg gagagctcca
     4321 aaagggaaac gagctcgccc tcccatccaa gtacgtcaac ttcctctacc tcgcctccca
     4381 ctacgagaag ctcaagggat ccccagagga caacgagcaa aagcaactct tcgtcgagca
     4441 acacaagcac tacctcgacg agatcatcga gcaaatctcc gagttctcca agcgtgtcat
     4501 cctcgccgac gccaacctcg acaaggtcct ctccgcctac aacaagcacc gtgacaagcc
     4561 aatccgtgag caagccgaga acatcatcca cctcttcacc ctcaccaacc tcggagcccc
     4621 agccgccttc aagtacttcg acaccaccat cgaccgtaag cgttacacct ccaccaaggt
     4681 aagttcataa gagaccttta ctaactaact aggtctcaga atttttcagg aggtcctcga
     4741 cgccaccctc atccaccaat ccatcaccgg actctacgag actcgtatcg acctctccca
     4801 actcggagga gacggaggat ccggaccacc aaagaagaag cgtaaggtct acccatacga
     4861 cgtcccagac tacgcctaaa cccagctttc ttgtacaaag tggcctaagc tgtctcatcc
     4921 tactttcacc tagttaactg cttgtcttaa aatctatgct tctctttagt atctaaaatt
     4981 ttcctagaag cttacaagta tataaatggt ctcttctcaa taaaggttgt atatttattc
     5041 atcttattga atctgccatt tcctcgtttt tgcgagttta tataccttcc aattttcttt
     5101 ctattgtatt ttcaacttct aattttaatt cagggaaact gcttcaacgc atcatgggtt
     5161 gtaccatgtc acaggaagag cgtgccgctc ttgaaagatc acgaatgtcc tcagtctcca
     5221 aggtaacact tagtttttgt tgctctgttt aaaaattaat tttaagggag aggagctcat
     5281 caaggagaac atgcacatga agctctacat ggagggaacc gtcaacaacc accacttcaa
     5341 gtgcacctcc gagggagagg gaaagccata cgagggaacc caaaccatgc gtatcaaggt
     5401 cgtcgaggga ggaccactcc cattcgcctt cgacatcctc gccacctcct tcatgtacgg
     5461 atcgcgtacc ttcatcaacc acacccaagg taagttttct tatgggaaag aaggaaaaaa
     5521 ccgagatttt acttgaaaaa ttgaattttt cgcgggattt tcaccaaaaa ttgttgaata
     5581 ttcattattt cacgctgtaa aacaaaaaaa aaaaaaatca aaaactacgt tgaaatcgcg
     5641 tttttaagcg aattttcttc agaattgcca gattttaacc ccaaattttg cagtttttaa
     5701 ataaaatttc accttttcgg ctcaaattgt agattttcct gaaaatttag tacaaaaaac
     5761 aatttcctcg taaatttttc aaatagattt tcaggaatcc cagacttctt caagcaatcc
     5821 ttcccagagg gattcacctg ggagcgtgtc accacctacg aggacggagg agtcctcacc
     5881 gccacccaag acacctccct ccaagacgga tgcctcatct acaacgtcaa ggtaagttat
     5941 gtctacctgc ctgcctaccg cctaaatttt ttgtgaagtt tcttcaaaaa atccagaaaa
     6001 aaaaacaatt ttcatacgat tttttccctt aaaattgtga attttcatgc tttttagccc
     6061 caaaagtcat tatttgagaa aaaatttcat acaaaaaagt tttgagaaat acacaatttt
     6121 ttaaatgtaa ttttcaaatt ttcaattttc aactagaaaa ttcacaaaac ttgtaaattt
     6181 tggaccaaaa catttataca attacttttt ttgaatctaa taactacaat aactacaaat
     6241 ttaacatttt cagatccgtg gagtcaactt cccatccaac ggaccagtca tgcaaaagaa
     6301 gaccctcgga tgggaggcca acaccgagat gctctaccca gccgacggag gactggaggg
     6361 acgtaccgac atggccctca agctcgtcgg aggaggacac ctcatctgca acttcaagac
     6421 cacctaccgt tccaagaagc cagccaaggt aagttctttt tttgaaaagt gcagtttgta
     6481 gtctaatttt catttatttt ctttttaaaa aacgcatcaa tttttaaata ttttgggaca
     6541 aaatccgaaa actgtactaa tttgtagttt gtaaaattaa aaaaaaaacg caaaaaaatg
     6601 ttttcaaaaa tgcttagaat aaaaataaaa tttctaaaaa attgaacaaa ataaacattt
     6661 aaaaattcaa aaagtttgaa aaaatgcaaa gttttttaat gcaaaaaata tttaatgttt
     6721 aaacaaattt aaaaaatgtt gtaagaatat gtttagagac agtttttaaa tgttaaattg
     6781 caaaaaaaaa aagcaaaaaa aaatttgaaa aatgtaacag aaaattttaa ttaaaaaagt
     6841 cgtgtttttt tcaaacctgc aattctttgt aattttattt ataaagagaa acaaatcaaa
     6901 ttgtatatta cgaaaaacga tactaaaatt cgaaaatttg cggttttttg cgtaaaaaat
     6961 acggttccgt aattttcaga acctcaagat gccaggagtc tactacgtcg accaccgtct
     7021 ggagcgtatc aaggaggccg acaaggagac gtacgtcgag caacacgagg tcgccgtcgc
     7081 ccgttactgc gacctcccat ccaagctcgg acacaagctc aacggaatgg acgagctcta
     7141 caagtcggga taattttccc cagatttccc cacgattttt tttgacattt ttatgaaaat
     7201 tttctctaaa agttacccct attttgttta gactgatctc ttattttatc caatgtttca
     7261 aatttttgaa attgctcgaa aattccaatt ttcacatatt tttgttgatt ttttcgccat
     7321 tttcccctga taaattaact tatttttgca attttttctt tcgaaaaaat attggtttct
     7381 catcaccatt tatccttttt tttatggagt taaaaattgt taaaaaaatt taattttttt
     7441 tctaatttta gagtcaaaac gggttcgaat ctagagcgta gctgtacgat tttttggcga
     7501 tttttgcggt tttctggtga gtttttttaa agttattttt ttgaaaaaaa ttcttgaaaa
     7561 taaaaggaaa ttgttttaaa aaaaaatttc aaagaaaaaa ttttgaaaaa attgaaacat
     7621 tttttttatt ttgaccaaaa atattgaaaa ttaattaaca tttttttttg gaaagttaat
     7681 ttttaataca aattttttaa atttatttta caaaaaaaat tgaatttctg tgaaaaatca
     7741 gttttccagt acataatgat ttgaccaaaa aaaattgaaa aaaatgattt tttaaaatta
     7801 aaaataccat aaaatgctga aaattcagaa aaataagtaa attttaaaaa attctggaaa
     7861 aacatctaaa aaccccccaa atataatatt ttggaccctt ttattttctg ctcgacatct
     7921 caaatatcct ccaatttttc gcttttttga ttaattgaaa gcatagtggg ggcaggtgtt
     7981 ataggacacc cttccctcca cttttttgga gcatttttca aagctcttcc gagagcagaa
     8041 gcttctcaat ttttccaccc caaaagtttg aaattctgcc aaaaaatctc gcattttaac
     8101 aatttttcga gatgtttcac gtccttttca ttgcgatttc ccaactttat tatacatagt
     8161 tgataattca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca
     8221 acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg
     8281 caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatggcgcc tgatgcggta
     8341 ttttctcctt acgcatctgt gcggtatttc acaccgcata tggtgcactc tcagtgcaat
     8401 ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc
     8461 ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag
     8521 ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt
     8581 gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg
     8641 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     8701 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     8761 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     8821 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     8881 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     8941 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     9001 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     9061 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     9121 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     9181 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     9241 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     9301 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     9361 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     9421 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     9481 ctctcgcggt atcattgcag cactggggtc agatggtaag ccctcccgta tcgtagttat
     9541 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     9601 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     9661 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     9721 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     9781 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     9841 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     9901 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     9961 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
    10021 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
    10081 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
    10141 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
    10201 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
    10261 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
    10321 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
    10381 gaaaaacgcc agcaacgcgg cctttttacg gttcctggtc ttttgctggc cttttgctca
    10441 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
    10501 agctgata
//
