LOCUS       sequence-136590-        5621 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  SapTrap GFP + Cbr-unc-119 donor plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5621)
  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 5621)
  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: pMLS252
FEATURES             Location/Qualifiers
     source          1..5621
                     /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"
     primer_bind     complement(707..735)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     protein_bind    1294..1327
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             3291..3302
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     protein_bind    3451..3484
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     3674..3694
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     protein_bind    complement(3746..3845)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3862..3881)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3863..3881)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3886..3902)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3886..3902)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4004..4023)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4015..4824
                     /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(4082..4101)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4971..5559
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5460..5479
                     /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 gcaaaggaga agaacttttc actggagttg tcccaattct
      721 tgttgaatta gatggtgatg ttaatgggca caaattttct gtcagtggag agggtgaagg
      781 tgatgcaaca tacggaaaac ttacccttaa atttatttgc actactggaa aactacctgt
      841 tccatgggta agtttaaaca tatatatact aactaaccct gattatttaa attttcagcc
      901 aacacttgtc actactttct gttatggtgt tcaatgcttc tcgagatacc cagatcatat
      961 gaaacggcat gactttttca agagtgccat gcccgaaggt tatgtacagg aaagaactat
     1021 atttttcaaa gatgacggga actacaagac acgtaagttt aaacagttcg gtactaacta
     1081 accatacata tttaaatttt caggtgctga agtcaagttt gaaggtgata cccttgttaa
     1141 tagaatcgag ttaaaaggta ttgattttaa agaagatgga aacattcttg gacacaaatt
     1201 ggaatacaac tataactcac acaatgtata catcatggca gacaaacaaa agaatggaat
     1261 caaagttgta agtttaaaca tgattttact aacataactt cgtataatgt atgctatacg
     1321 aagttatttc tagacattct ctaatgaaaa aatctttcag ttgaaattga aaatgagtta
     1381 aagttggagt ttttattgaa aacagatttc cgtgtgatta gtgtttttag cgagtgtgac
     1441 aggacagcga aaaaatatag aaacaagggg ggaactgaaa agcttaggaa tgcattgaac
     1501 atgagaaggg gaaggggaag gaacaaacta gacaggaatt attggaattt aatcacattt
     1561 ggagtttttt ttctattcga cagaataatt atccagaaca tttttgtatt aaatatttat
     1621 gcatcatatg agtagtcggc tttgttgtgc atgacgagtt tgttatcgac gaaatagaag
     1681 ctgtcagaac gagtctcgtt tggattgttg atcatgtcgt ccactgaaaa agagattagt
     1741 cttttgaatt gtacttttta gaataatgac tcactgagtt gttgagagag ttgagggaac
     1801 tcatagatat gttcacagtt gtttcgtgaa ttcggaatac agaatccgaa ttcaaagtca
     1861 aaacacttca gaaggcgatc tttgaagaag tgacgttcga tcattcggaa atgatggatt
     1921 gggatgtctc ctactttgaa ttccacagtt gctccgaccg ttttgagttt caaaaagttt
     1981 ggagcaaatc tataacgcac gtatcttgcc gactcctgtg gcgactcatc attctcctga
     2041 tcgttctccg gtttggcgat ctcgaaaagc acttgctcag tgtccagatc acggatttgg
     2101 aacttggtga actcgatgtt atagatgttc gcagatgggg agcataagaa tcctaaattt
     2161 atgttttaaa ctgaaatcca aagggagcaa gataccttga gtgattcccg gaagtgctaa
     2221 aacgtcgttc ggagtgattt gagctttctt cgcaagctcc gattccgttg tgattccttg
     2281 ttcggtgctt ggtggtggcc gtggcatctg gaaatatgga aaagttcaac aaaaagaaaa
     2341 gagaaaagaa tgaaatcgga tatcaagagt tagttgagcg gtttctctag ttttctgagt
     2401 ctcacctgcg acgggaaggt cgccgagccg ggtggaatcg attgttgttg ctcggctttc
     2461 atatcggttt ggttggaagc ggctgaaaac ggaaagaagt ggaagaagga aaagagtgtg
     2521 gtgtgacagg aaaatggtaa ttagagggtg ccaaataacc agctatattt tgtttttttt
     2581 tgaaaacatt tttaaaaaga aaaatacgat aatgatatca gatggatttc cggaaaactg
     2641 gtatgaaaaa tttcaacctt tttgagtaca tgtaatcaaa atacactttg taaattatca
     2701 tttttattga aactccacca tttttctatt tataacgcta ataatttgaa aaagaaacct
     2761 gttgcgaacc gcggggtgaa tcccaaaaac gaatgcgttt tggtggagtg attgattcga
     2821 atcgaagaag aaaaagaaga agacgtggaa tagagagctc actcttaacc gagcagcaca
     2881 caccgacaga aaaaaaaatg aaatgaatga gggtcttctt cttcttcttc ttcgaatgat
     2941 tgacagaaat gggaaaaaga ggaagattga gaagggaaaa aggaaggaga aaagaagcag
     3001 aagaagacgt cagagaggag aggaacgagc ggaaaagcag cgggcgcaag tcatagaagt
     3061 agcagagctg gggagaagaa gacactatcc aagaaaggaa tgacgagaga gtatgcaaag
     3121 gggtataggg tgcagacaga ataggaacag aataacagat gatgagccaa gaagagttga
     3181 aaagggcgat gaatttgtca tgtaacttaa tttgggtcaa tttgagcatg atgaattgaa
     3241 atcatccctt gttgggagtt aataaccggt ttgttatcag aaaccctgta atagaagggc
     3301 gccctaactt tgagccaatt catcccggtt tctgtcaaat atatcaaaaa gtggtcaact
     3361 gacaaattgt ttttgatatt ataataaaca ttttatccgt taacaatttt cgaatacttt
     3421 ttacaaggac ttggataaat tggctcaaag ataacttcgt ataatgtatg ctatacgaag
     3481 ttattaacta atctgattta aattttcaga acttcaaaat tagacacaac attgaagatg
     3541 gaagcgttca actagcagac cattatcaac aaaatactcc aattggcgat ggccctgtcc
     3601 ttttaccaga caaccattac ctgtccacac aatctgccct ttcgaaagat cccaacgaaa
     3661 agagagacca catggtcctt cttgagtttg taacagctgc tgggattaca catggcatgg
     3721 atgaactata caaaggtcga agagcaccca gctttcttgt acaaagttgg cattataaga
     3781 aagcattgct tatcaatttg ttgcaacgaa caggtcacta tcagtcaaaa taaaatcatt
     3841 atttgccatc cagctgatat cccctatagt gagtcgtatt acatggtcat agctgtttcc
     3901 tggcagctct ggcccgtgtc tcaaaatctc tgatgttaca ttgcacaaga taaaataata
     3961 tcatcatgaa caataaaact gtctgcttac ataaacagta atacaagggg tgttatgagc
     4021 catattcaac gggaaacgtc gaggccgcga ttaaattcca acatggatgc tgatttatat
     4081 gggtataaat gggctcgcga taatgtcggg caatcaggtg cgacaatcta tcgcttgtat
     4141 gggaagcccg atgcgccaga gttgtttctg aaacatggca aaggtagcgt tgccaatgat
     4201 gttacagatg agatggtcag actaaactgg ctgacggaat ttatgcctct tccgaccatc
     4261 aagcatttta tccgtactcc tgatgatgca tggttactca ccactgcgat ccccggaaaa
     4321 acagcattcc aggtattaga agaatatcct gattcaggtg aaaatattgt tgatgcgctg
     4381 gcagtgttcc tgcgccggtt gcattcgatt cctgtttgta attgtccttt taacagcgat
     4441 cgcgtatttc gtctcgctca ggcgcaatca cgaatgaata acggtttggt tgatgcgagt
     4501 gattttgatg acgagcgtaa tggctggcct gttgaacaag tctggaaaga aatgcataaa
     4561 cttttgccat tctcaccgga ttcagtcgtc actcatggtg atttctcact tgataacctt
     4621 atttttgacg aggggaaatt aataggttgt attgatgttg gacgagtcgg aatcgcagac
     4681 cgataccagg atcttgccat cctatggaac tgcctcggtg agttttctcc ttcattacag
     4741 aaacggcttt ttcaaaaata tggtattgat aatcctgata tgaataaatt gcagtttcat
     4801 ttgatgctcg atgagttttt ctaatcagaa ttggttaatt ggttgtaaca ctggcagagc
     4861 attacgctga cttgacggga cggcgcaagc tcatgaccaa aatcccttaa cgtgagttac
     4921 gcgtcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     4981 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     5041 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     5101 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     5161 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     5221 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     5281 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     5341 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     5401 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     5461 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     5521 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     5581 ttacggttcc tggccttttg ctggcctttt gctcacatgt t
//
