LOCUS       sequence-136592-        5699 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  SapTrap Halotag + Cbr-unc-119 donor plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5699)
  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 5699)
  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: pMLS254
FEATURES             Location/Qualifiers
     source          1..5699
                     /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    1217..1250
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             3214..3225
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     protein_bind    3374..3407
                     /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(3824..3923)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3940..3959)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3941..3959)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3964..3980)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3964..3980)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4082..4101)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4093..4902
                     /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(4160..4179)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      5049..5637
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5538..5557
                     /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 tggctgaaat tggtactggc tttccttttg atccccacta
      721 cgttgaggtt cttggagagc gcatgcacta cgttgatgtt ggaccacgcg atggaacccc
      781 agtccttttc cttcacggaa acccaacttc ttcttacgtt tggcgcaaca tcatcccaca
      841 tgttgctcca acccaccgct gcatcgctcc agatcttatc ggaatgggaa agtctgataa
      901 gccagatctt ggatacttct tcgatgatca cgtccgtttc atggatgctt tcatcgaggc
      961 ccttggactt gaggaggttg tccttgttat ccacgattgg ggatccgctc ttggattcca
     1021 ctgggctaag cgcaacccag agcgcgttaa gggaatcgct ttcatggaat tcatccgccc
     1081 aatcccaact tgggatgaat ggccagagtt cgctcgcgag accttccaag ctttccgcac
     1141 caccgatgtt ggacgtaagc ttatcatcga tcaaaacgtt ttcatcgaag gtaagtttaa
     1201 acatgatttt actaacataa cttcgtataa tgtatgctat acgaagttat ttctagacat
     1261 tctctaatga aaaaatcttt cagttgaaat tgaaaatgag ttaaagttgg agtttttatt
     1321 gaaaacagat ttccgtgtga ttagtgtttt tagcgagtgt gacaggacag cgaaaaaata
     1381 tagaaacaag gggggaactg aaaagcttag gaatgcattg aacatgagaa ggggaagggg
     1441 aaggaacaaa ctagacagga attattggaa tttaatcaca tttggagttt tttttctatt
     1501 cgacagaata attatccaga acatttttgt attaaatatt tatgcatcat atgagtagtc
     1561 ggctttgttg tgcatgacga gtttgttatc gacgaaatag aagctgtcag aacgagtctc
     1621 gtttggattg ttgatcatgt cgtccactga aaaagagatt agtcttttga attgtacttt
     1681 ttagaataat gactcactga gttgttgaga gagttgaggg aactcataga tatgttcaca
     1741 gttgtttcgt gaattcggaa tacagaatcc gaattcaaag tcaaaacact tcagaaggcg
     1801 atctttgaag aagtgacgtt cgatcattcg gaaatgatgg attgggatgt ctcctacttt
     1861 gaattccaca gttgctccga ccgttttgag tttcaaaaag tttggagcaa atctataacg
     1921 cacgtatctt gccgactcct gtggcgactc atcattctcc tgatcgttct ccggtttggc
     1981 gatctcgaaa agcacttgct cagtgtccag atcacggatt tggaacttgg tgaactcgat
     2041 gttatagatg ttcgcagatg gggagcataa gaatcctaaa tttatgtttt aaactgaaat
     2101 ccaaagggag caagatacct tgagtgattc ccggaagtgc taaaacgtcg ttcggagtga
     2161 tttgagcttt cttcgcaagc tccgattccg ttgtgattcc ttgttcggtg cttggtggtg
     2221 gccgtggcat ctggaaatat ggaaaagttc aacaaaaaga aaagagaaaa gaatgaaatc
     2281 ggatatcaag agttagttga gcggtttctc tagttttctg agtctcacct gcgacgggaa
     2341 ggtcgccgag ccgggtggaa tcgattgttg ttgctcggct ttcatatcgg tttggttgga
     2401 agcggctgaa aacggaaaga agtggaagaa ggaaaagagt gtggtgtgac aggaaaatgg
     2461 taattagagg gtgccaaata accagctata ttttgttttt ttttgaaaac atttttaaaa
     2521 agaaaaatac gataatgata tcagatggat ttccggaaaa ctggtatgaa aaatttcaac
     2581 ctttttgagt acatgtaatc aaaatacact ttgtaaatta tcatttttat tgaaactcca
     2641 ccatttttct atttataacg ctaataattt gaaaaagaaa cctgttgcga accgcggggt
     2701 gaatcccaaa aacgaatgcg ttttggtgga gtgattgatt cgaatcgaag aagaaaaaga
     2761 agaagacgtg gaatagagag ctcactctta accgagcagc acacaccgac agaaaaaaaa
     2821 atgaaatgaa tgagggtctt cttcttcttc ttcttcgaat gattgacaga aatgggaaaa
     2881 agaggaagat tgagaaggga aaaaggaagg agaaaagaag cagaagaaga cgtcagagag
     2941 gagaggaacg agcggaaaag cagcgggcgc aagtcataga agtagcagag ctggggagaa
     3001 gaagacacta tccaagaaag gaatgacgag agagtatgca aaggggtata gggtgcagac
     3061 agaataggaa cagaataaca gatgatgagc caagaagagt tgaaaagggc gatgaatttg
     3121 tcatgtaact taatttgggt caatttgagc atgatgaatt gaaatcatcc cttgttggga
     3181 gttaataacc ggtttgttat cagaaaccct gtaatagaag ggcgccctaa ctttgagcca
     3241 attcatcccg gtttctgtca aatatatcaa aaagtggtca actgacaaat tgtttttgat
     3301 attataataa acattttatc cgttaacaat tttcgaatac tttttacaag gacttggata
     3361 aattggctca aagataactt cgtataatgt atgctatacg aagttattaa ctaatctgat
     3421 ttaaattttc aggaaccctt ccaatgggag ttgttcgtcc acttaccgag gttgagatgg
     3481 atcactaccg cgagccattc cttaacccag ttgatcgcga gccactttgg cgcttcccaa
     3541 acgaacttcc aatcgctgga gagccagcta acatcgttgc tcttgttgag gagtacatgg
     3601 attggcttca ccaatctcca gttccaaagc ttcttttctg gggaactcca ggagttctta
     3661 tcccaccagc tgaggctgcc cgtcttgcta agtctcttcc aaactgcaag gctgtcgata
     3721 tcggaccagg acttaacctt cttcaagagg ataacccaga tcttatcgga tctgagatcg
     3781 ctcgttggct ttctaccctt gagatctccg gaggtcgaag agcacccagc tttcttgtac
     3841 aaagttggca ttataagaaa gcattgctta tcaatttgtt gcaacgaaca ggtcactatc
     3901 agtcaaaata aaatcattat ttgccatcca gctgatatcc cctatagtga gtcgtattac
     3961 atggtcatag ctgtttcctg gcagctctgg cccgtgtctc aaaatctctg atgttacatt
     4021 gcacaagata aaataatatc atcatgaaca ataaaactgt ctgcttacat aaacagtaat
     4081 acaaggggtg ttatgagcca tattcaacgg gaaacgtcga ggccgcgatt aaattccaac
     4141 atggatgctg atttatatgg gtataaatgg gctcgcgata atgtcgggca atcaggtgcg
     4201 acaatctatc gcttgtatgg gaagcccgat gcgccagagt tgtttctgaa acatggcaaa
     4261 ggtagcgttg ccaatgatgt tacagatgag atggtcagac taaactggct gacggaattt
     4321 atgcctcttc cgaccatcaa gcattttatc cgtactcctg atgatgcatg gttactcacc
     4381 actgcgatcc ccggaaaaac agcattccag gtattagaag aatatcctga ttcaggtgaa
     4441 aatattgttg atgcgctggc agtgttcctg cgccggttgc attcgattcc tgtttgtaat
     4501 tgtcctttta acagcgatcg cgtatttcgt ctcgctcagg cgcaatcacg aatgaataac
     4561 ggtttggttg atgcgagtga ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc
     4621 tggaaagaaa tgcataaact tttgccattc tcaccggatt cagtcgtcac tcatggtgat
     4681 ttctcacttg ataaccttat ttttgacgag gggaaattaa taggttgtat tgatgttgga
     4741 cgagtcggaa tcgcagaccg ataccaggat cttgccatcc tatggaactg cctcggtgag
     4801 ttttctcctt cattacagaa acggcttttt caaaaatatg gtattgataa tcctgatatg
     4861 aataaattgc agtttcattt gatgctcgat gagtttttct aatcagaatt ggttaattgg
     4921 ttgtaacact ggcagagcat tacgctgact tgacgggacg gcgcaagctc atgaccaaaa
     4981 tcccttaacg tgagttacgc gtcgttccac tgagcgtcag accccgtaga aaagatcaaa
     5041 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     5101 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     5161 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     5221 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     5281 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     5341 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     5401 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     5461 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     5521 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     5581 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     5641 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgtt
//
