LOCUS       sequence-140083-        5348 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  SapTrap SNAPtag + Cbr-unc-119 donor plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5348)
  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 5348)
  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: pMLS271
FEATURES             Location/Qualifiers
     source          1..5348
                     /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"
     protein_bind    977..1010
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             2974..2985
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     protein_bind    3134..3167
                     /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(3473..3572)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3589..3608)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3590..3608)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3613..3629)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3613..3629)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3731..3750)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3742..4551
                     /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(3809..3828)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4698..5286
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5187..5206
                     /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 tcttcgatgg acaaggactg cgagatgaag cgtaccaccc tcgactcccc
      721 actcggaaag ctcgagctct ccggatgcga gcaaggactc caccgtatca tcttcctcgg
      781 aaagggaacc tccgccgccg acgccgtcga ggtcccagcc ccagccgccg tcctcggagg
      841 accagagcca ctcatgcaag ccaccgcctg gctcaacgcc tacttccacc aaccagaggc
      901 catcgaggag ttcccagtcc cagccctcca ccacccagtc ttccaacaag gtaagtttaa
      961 acatgatttt actaacataa cttcgtataa tgtatgctat acgaagttat ttctagacat
     1021 tctctaatga aaaaatcttt cagttgaaat tgaaaatgag ttaaagttgg agtttttatt
     1081 gaaaacagat ttccgtgtga ttagtgtttt tagcgagtgt gacaggacag cgaaaaaata
     1141 tagaaacaag gggggaactg aaaagcttag gaatgcattg aacatgagaa ggggaaaggg
     1201 aaggaacaaa ctagacagga attattggaa tttaatcaca tttggagttt tttttctatt
     1261 cgacagaata attatccaga acatttttgt attaaatatt tatgcatcat atgagtagtc
     1321 ggctttgttg tgcatgacga gtttgttatc gacgaaatag aagctgtcag aacgagtctc
     1381 gtttggattg ttgatcatgt cgtccactga aaaagagatt agtcttttga attgtacttt
     1441 ttagaataat gactcactga gttgttgaga gagttgaggg aactcataga tatgttcaca
     1501 gttgtttcgt gaattcggaa tacagaatcc gaattcaaag tcaaaacact tcagaaggcg
     1561 atctttgaag aagtgacgtt cgatcattcg gaaatgatgg attgggatgt ctcctacttt
     1621 gaattccaca gttgctccga ccgttttgag tttcaaaaag tttggagcaa atctataacg
     1681 cacgtatctt gccgactcct gtggcgactc atcattctcc tgatcgttct ccggtttggc
     1741 gatctcgaaa agcacttgct cagtgtccag atcacggatt tggaacttgg tgaactcgat
     1801 gttatagatg ttcgcagatg gggagcataa gaatcctaaa tttatgtttt aaactgaaat
     1861 ccaaagggag caagatacct tgagtgattc ccggaagtgc taaaacgtcg ttcggagtga
     1921 tttgagcttt cttcgcaagc tccgattccg ttgtgattcc ttgttcggtg cttggtggtg
     1981 gccgtggcat ctggaaatat ggaaaagttc aacaaaaaga aaagagaaaa gaatgaaatc
     2041 ggatatcaag agttagttga gcggtttctc tagttttctg agtctcacct gcgacgggaa
     2101 ggtcgccgag ccgggtggaa tcgattgttg ttgctcggct ttcatatcgg tttggttgga
     2161 agcggctgaa aacggaaaga agtggaagaa ggaaaagagt gtggtgtgac aggaaaatgg
     2221 taattagagg gtgccaaata accagctata ttttgttttt ttttgaaaac atttttaaaa
     2281 agaaaaatac gataatgata tcagatggat ttccggaaaa ctggtatgaa aaatttcaac
     2341 ctttttgagt acatgtaatc aaaatacact ttgtaaatta tcatttttat tgaaactcca
     2401 ccatttttct atttataacg ctaataattt gaaaaagaaa cctgttgcga accgcggggt
     2461 gaatcccaaa aacgaatgcg ttttggtgga gtgattgatt cgaatcgaag aagaaaaaga
     2521 agaagacgtg gaatagagag ctcactctta accgagcagc acacaccgac agaaaaaaaa
     2581 atgaaatgaa tgagggtctt cttcttcttc ttcttcgaat gattgacaga aatgggaaaa
     2641 agaggaagat tgagaaggga aaaaggaagg agaaaagaag cagaagaaga cgtcagagag
     2701 gagaggaacg agcggaaaag cagcgggcgc aagtcataga agtagcagag ctggggagaa
     2761 gaagacacta tccaagaaag gaatgacgag agagtatgca aaggggtata gggtgcagac
     2821 agaataggaa cagaataaca gatgatgagc caagaagagt tgaaaagggc gatgaatttg
     2881 tcatgtaact taatttgggt caatttgagc atgatgaatt gaaatcatcc cttgttggga
     2941 gttaataacc ggtttgttat cagaaaccct gtaatagaag ggcgccctaa ctttgagcca
     3001 attcatcccg gtttctgtca aatatatcaa aaagtggtca actgacaaat tgtttttgat
     3061 attataataa acattttatc cgttaacaat tttcgaatac tttttacaag gacttggata
     3121 aattggctca aagataactt cgtataatgt atgctatacg aagttattaa ctaatctgat
     3181 ttaaattttc agagtccttc acccgtcaag tcctctggaa gctcctcaag gtcgtcaagt
     3241 tcggagaggt catctcctac tcccacctcg ccgccctcgc cggaaaccca gccgccaccg
     3301 ccgccgtcaa gaccgccctc tccggaaacc cagtcccaat cctcatccca tgccaccgtg
     3361 tcgtccaagg agacctcgac gtcggaggat acgagggagg actcgccgtc aaggagtggc
     3421 tcctcgccca cgagggacac cgtctcggaa agccaggact cggtcgaaga gcacccagct
     3481 ttcttgtaca aagttggcat tataagaaag cattgcttat caatttgttg caacgaacag
     3541 gtcactatca gtcaaaataa aatcattatt tgccatccag ctgatatccc ctatagtgag
     3601 tcgtattaca tggtcatagc tgtttcctgg cagctctggc ccgtgtctca aaatctctga
     3661 tgttacattg cacaagataa aataatatca tcatgaacaa taaaactgtc tgcttacata
     3721 aacagtaata caaggggtgt tatgagccat attcaacggg aaacgtcgag gccgcgatta
     3781 aattccaaca tggatgctga tttatatggg tataaatggg ctcgcgataa tgtcgggcaa
     3841 tcaggtgcga caatctatcg cttgtatggg aagcccgatg cgccagagtt gtttctgaaa
     3901 catggcaaag gtagcgttgc caatgatgtt acagatgaga tggtcagact aaactggctg
     3961 acggaattta tgcctcttcc gaccatcaag cattttatcc gtactcctga tgatgcatgg
     4021 ttactcacca ctgcgatccc cggaaaaaca gcattccagg tattagaaga atatcctgat
     4081 tcaggtgaaa atattgttga tgcgctggca gtgttcctgc gccggttgca ttcgattcct
     4141 gtttgtaatt gtccttttaa cagcgatcgc gtatttcgtc tcgctcaggc gcaatcacga
     4201 atgaataacg gtttggttga tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt
     4261 gaacaagtct ggaaagaaat gcataaactt ttgccattct caccggattc agtcgtcact
     4321 catggtgatt tctcacttga taaccttatt tttgacgagg ggaaattaat aggttgtatt
     4381 gatgttggac gagtcggaat cgcagaccga taccaggatc ttgccatcct atggaactgc
     4441 ctcggtgagt tttctccttc attacagaaa cggctttttc aaaaatatgg tattgataat
     4501 cctgatatga ataaattgca gtttcatttg atgctcgatg agtttttcta atcagaattg
     4561 gttaattggt tgtaacactg gcagagcatt acgctgactt gacgggacgg cgcaagctca
     4621 tgaccaaaat cccttaacgt gagttacgcg tcgttccact gagcgtcaga ccccgtagaa
     4681 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     4741 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     4801 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     4861 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     4921 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     4981 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     5041 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     5101 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     5161 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     5221 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     5281 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     5341 cacatgtt
//
