LOCUS       sequence-136600-        5747 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  SapTrap 2xNLS-mCherry + Cbr-unc-119 donor plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5747)
  AUTHORS   Schwartz ML, Jorgensen EM
  TITLE     SapTrap, a Toolkit for High-Throughput CRISPR/Cas9 Gene Modification
            in Caenorhabditis elegans.
  JOURNAL   Genetics. 2016 Feb 2. pii: genetics.115.184275.
  PUBMED    26837755
REFERENCE   2  (bases 1 to 5747)
  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: "Genetics. 
            2016 Feb 2. pii: genetics.115.184275."
COMMENT     Sequence Label: pMLS292
FEATURES             Location/Qualifiers
     source          1..5747
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     536..553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    569..668
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             686..706
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     protein_bind    1393..1426
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             3390..3401
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     protein_bind    3550..3583
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    complement(3872..3971)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3988..4007)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3989..4007)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4012..4028)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4012..4028)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4130..4149)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4141..4950
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(4208..4227)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      5097..5685
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5586..5605
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac
      601 ctgttcgttg caacaaattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
      661 agcaggctgc tcttcgatga ctgctccaaa gaagaagcgt aaggtaccgg tagaaaaagg
      721 gatggtctca aagggtgaag aagataacat ggcaattatt aaagagttta tgcgtttcaa
      781 ggtgcatatg gagggatctg tcaatgggca tgagtttgaa attgaaggtg aaggagaagg
      841 ccgaccatat gagggaacac aaaccgcaaa actaaaggta agtttaaaca tatatatact
      901 aactaaccct gattatttaa attttcaggt aactaaaggc ggaccattac cattcgcctg
      961 ggacatcctc tctccacagt tcatgtatgg aagtaaagct tatgttaaac atccggcaga
     1021 tataccagat tatttgaaac tttcattccc ggagggtttt aagtgggaac gcgtaatgaa
     1081 ttttgaagac ggaggagttg ttacagtgac gcaagactca aggtaagttt aaacagttcg
     1141 gtactaacta accatacata tttaaatttt cagcctccaa gatggagaat ttatttataa
     1201 agtcaaactt cgaggaacga atttcccctc ggatggacct gttatgcaga agaagactat
     1261 gggatgggaa gcttcaagtg aaagaatgta ccctgaagac ggtgctctta agggagagat
     1321 taaacaacgt cttaaattga aagatggagg acattacgat gctgaggtaa gtttaaacat
     1381 gattttacta acataacttc gtataatgta tgctatacga agttatttct agacattctc
     1441 taatgaaaaa atctttcagt tgaaattgaa aatgagttaa agttggagtt tttattgaaa
     1501 acagatttcc gtgtgattag tgtttttagc gagtgtgaca ggacagcgaa aaaatataga
     1561 aacaaggggg gaactgaaaa gcttaggaat gcattgaaca tgagaagggg aaggggaagg
     1621 aacaaactag acaggaatta ttggaattta atcacatttg gagttttttt tctattcgac
     1681 agaataatta tccagaacat ttttgtatta aatatttatg catcatatga gtagtcggct
     1741 ttgttgtgca tgacgagttt gttatcgacg aaatagaagc tgtcagaacg agtctcgttt
     1801 ggattgttga tcatgtcgtc cactgaaaaa gagattagtc ttttgaattg tactttttag
     1861 aataatgact cactgagttg ttgagagagt tgagggaact catagatatg ttcacagttg
     1921 tttcgtgaat tcggaataca gaatccgaat tcaaagtcaa aacacttcag aaggcgatct
     1981 ttgaagaagt gacgttcgat cattcggaaa tgatggattg ggatgtctcc tactttgaat
     2041 tccacagttg ctccgaccgt tttgagtttc aaaaagtttg gagcaaatct ataacgcacg
     2101 tatcttgccg actcctgtgg cgactcatca ttctcctgat cgttctccgg tttggcgatc
     2161 tcgaaaagca cttgctcagt gtccagatca cggatttgga acttggtgaa ctcgatgtta
     2221 tagatgttcg cagatgggga gcataagaat cctaaattta tgttttaaac tgaaatccaa
     2281 agggagcaag ataccttgag tgattcccgg aagtgctaaa acgtcgttcg gagtgatttg
     2341 agctttcttc gcaagctccg attccgttgt gattccttgt tcggtgcttg gtggtggccg
     2401 tggcatctgg aaatatggaa aagttcaaca aaaagaaaag agaaaagaat gaaatcggat
     2461 atcaagagtt agttgagcgg tttctctagt tttctgagtc tcacctgcga cgggaaggtc
     2521 gccgagccgg gtggaatcga ttgttgttgc tcggctttca tatcggtttg gttggaagcg
     2581 gctgaaaacg gaaagaagtg gaagaaggaa aagagtgtgg tgtgacagga aaatggtaat
     2641 tagagggtgc caaataacca gctatatttt gttttttttt gaaaacattt ttaaaaagaa
     2701 aaatacgata atgatatcag atggatttcc ggaaaactgg tatgaaaaat ttcaaccttt
     2761 ttgagtacat gtaatcaaaa tacactttgt aaattatcat ttttattgaa actccaccat
     2821 ttttctattt ataacgctaa taatttgaaa aagaaacctg ttgcgaaccg cggggtgaat
     2881 cccaaaaacg aatgcgtttt ggtggagtga ttgattcgaa tcgaagaaga aaaagaagaa
     2941 gacgtggaat agagagctca ctcttaaccg agcagcacac accgacagaa aaaaaaatga
     3001 aatgaatgag ggtcttcttc ttcttcttct tcgaatgatt gacagaaatg ggaaaaagag
     3061 gaagattgag aagggaaaaa ggaaggagaa aagaagcaga agaagacgtc agagaggaga
     3121 ggaacgagcg gaaaagcagc gggcgcaagt catagaagta gcagagctgg ggagaagaag
     3181 acactatcca agaaaggaat gacgagagag tatgcaaagg ggtatagggt gcagacagaa
     3241 taggaacaga ataacagatg atgagccaag aagagttgaa aagggcgatg aatttgtcat
     3301 gtaacttaat ttgggtcaat ttgagcatga tgaattgaaa tcatcccttg ttgggagtta
     3361 ataaccggtt tgttatcaga aaccctgtaa tagaagggcg ccctaacttt gagccaattc
     3421 atcccggttt ctgtcaaata tatcaaaaag tggtcaactg acaaattgtt tttgatatta
     3481 taataaacat tttatccgtt aacaattttc gaatactttt tacaaggact tggataaatt
     3541 ggctcaaaga taacttcgta taatgtatgc tatacgaagt tattaactaa tctgatttaa
     3601 attttcaggt gaagacaact tacaaagcca aaaaaccagt tcagctgcca ggagcgtaca
     3661 atgttaatat taaactggat atcacctccc acaacgagga ttacactatc gttgagcaat
     3721 atgaaagagc tgaagggcgg cactcgacag gtggcatgga tgaattgtat aaggggagct
     3781 ctatgagccg tagacgaaaa gcgaatccga caaaactgag tgaaaacgcg aagaagcttg
     3841 ccaaggaagt tgaaaattaa ggtcgaagag cacccagctt tcttgtacaa agttggcatt
     3901 ataagaaagc attgcttatc aatttgttgc aacgaacagg tcactatcag tcaaaataaa
     3961 atcattattt gccatccagc tgatatcccc tatagtgagt cgtattacat ggtcatagct
     4021 gtttcctggc agctctggcc cgtgtctcaa aatctctgat gttacattgc acaagataaa
     4081 ataatatcat catgaacaat aaaactgtct gcttacataa acagtaatac aaggggtgtt
     4141 atgagccata ttcaacggga aacgtcgagg ccgcgattaa attccaacat ggatgctgat
     4201 ttatatgggt ataaatgggc tcgcgataat gtcgggcaat caggtgcgac aatctatcgc
     4261 ttgtatggga agcccgatgc gccagagttg tttctgaaac atggcaaagg tagcgttgcc
     4321 aatgatgtta cagatgagat ggtcagacta aactggctga cggaatttat gcctcttccg
     4381 accatcaagc attttatccg tactcctgat gatgcatggt tactcaccac tgcgatcccc
     4441 ggaaaaacag cattccaggt attagaagaa tatcctgatt caggtgaaaa tattgttgat
     4501 gcgctggcag tgttcctgcg ccggttgcat tcgattcctg tttgtaattg tccttttaac
     4561 agcgatcgcg tatttcgtct cgctcaggcg caatcacgaa tgaataacgg tttggttgat
     4621 gcgagtgatt ttgatgacga gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg
     4681 cataaacttt tgccattctc accggattca gtcgtcactc atggtgattt ctcacttgat
     4741 aaccttattt ttgacgaggg gaaattaata ggttgtattg atgttggacg agtcggaatc
     4801 gcagaccgat accaggatct tgccatccta tggaactgcc tcggtgagtt ttctccttca
     4861 ttacagaaac ggctttttca aaaatatggt attgataatc ctgatatgaa taaattgcag
     4921 tttcatttga tgctcgatga gtttttctaa tcagaattgg ttaattggtt gtaacactgg
     4981 cagagcatta cgctgacttg acgggacggc gcaagctcat gaccaaaatc ccttaacgtg
     5041 agttacgcgt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     5101 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     5161 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     5221 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     5281 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     5341 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     5401 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     5461 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     5521 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     5581 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     5641 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     5701 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgtt
//
