LOCUS       sequence-136598-        5615 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  SapTrap mCherry + Cbr-unc-119 donor plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5615)
  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 5615)
  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: pMLS291
FEATURES             Location/Qualifiers
     source          1..5615
                     /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    1348..1381
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             3345..3356
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     protein_bind    3505..3538
                     /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(3740..3839)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3856..3875)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3857..3875)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3880..3896)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3880..3896)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3998..4017)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4009..4818
                     /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(4076..4095)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4965..5553
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5454..5473
                     /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 tctcaaaggg tgaagaagat aacatggcaa ttattaaaga
      721 gtttatgcgt ttcaaggtgc atatggaggg atctgtcaat gggcatgagt ttgaaattga
      781 aggtgaagga gaaggccgac catatgaggg aacacaaacc gcaaaactaa aggtaagttt
      841 aaacatatat atactaacta accctgatta tttaaatttt caggtaacta aaggcggacc
      901 attaccattc gcctgggaca tcctctctcc acagttcatg tatggaagta aagcttatgt
      961 taaacatccg gcagatatac cagattattt gaaactttca ttcccggagg gttttaagtg
     1021 ggaacgcgta atgaattttg aagacggagg agttgttaca gtgacgcaag actcaaggta
     1081 agtttaaaca gttcggtact aactaaccat acatatttaa attttcagcc tccaagatgg
     1141 agaatttatt tataaagtca aacttcgagg aacgaatttc ccctcggatg gacctgttat
     1201 gcagaagaag actatgggat gggaagcttc aagtgaaaga atgtaccctg aagacggtgc
     1261 tcttaaggga gagattaaac aacgtcttaa attgaaagat ggaggacatt acgatgctga
     1321 ggtaagttta aacatgattt tactaacata acttcgtata atgtatgcta tacgaagtta
     1381 tttctagaca ttctctaatg aaaaaatctt tcagttgaaa ttgaaaatga gttaaagttg
     1441 gagtttttat tgaaaacaga tttccgtgtg attagtgttt ttagcgagtg tgacaggaca
     1501 gcgaaaaaat atagaaacaa ggggggaact gaaaagctta ggaatgcatt gaacatgaga
     1561 aggggaaggg gaaggaacaa actagacagg aattattgga atttaatcac atttggagtt
     1621 ttttttctat tcgacagaat aattatccag aacatttttg tattaaatat ttatgcatca
     1681 tatgagtagt cggctttgtt gtgcatgacg agtttgttat cgacgaaata gaagctgtca
     1741 gaacgagtct cgtttggatt gttgatcatg tcgtccactg aaaaagagat tagtcttttg
     1801 aattgtactt tttagaataa tgactcactg agttgttgag agagttgagg gaactcatag
     1861 atatgttcac agttgtttcg tgaattcgga atacagaatc cgaattcaaa gtcaaaacac
     1921 ttcagaaggc gatctttgaa gaagtgacgt tcgatcattc ggaaatgatg gattgggatg
     1981 tctcctactt tgaattccac agttgctccg accgttttga gtttcaaaaa gtttggagca
     2041 aatctataac gcacgtatct tgccgactcc tgtggcgact catcattctc ctgatcgttc
     2101 tccggtttgg cgatctcgaa aagcacttgc tcagtgtcca gatcacggat ttggaacttg
     2161 gtgaactcga tgttatagat gttcgcagat ggggagcata agaatcctaa atttatgttt
     2221 taaactgaaa tccaaaggga gcaagatacc ttgagtgatt cccggaagtg ctaaaacgtc
     2281 gttcggagtg atttgagctt tcttcgcaag ctccgattcc gttgtgattc cttgttcggt
     2341 gcttggtggt ggccgtggca tctggaaata tggaaaagtt caacaaaaag aaaagagaaa
     2401 agaatgaaat cggatatcaa gagttagttg agcggtttct ctagttttct gagtctcacc
     2461 tgcgacggga aggtcgccga gccgggtgga atcgattgtt gttgctcggc tttcatatcg
     2521 gtttggttgg aagcggctga aaacggaaag aagtggaaga aggaaaagag tgtggtgtga
     2581 caggaaaatg gtaattagag ggtgccaaat aaccagctat attttgtttt tttttgaaaa
     2641 catttttaaa aagaaaaata cgataatgat atcagatgga tttccggaaa actggtatga
     2701 aaaatttcaa cctttttgag tacatgtaat caaaatacac tttgtaaatt atcattttta
     2761 ttgaaactcc accatttttc tatttataac gctaataatt tgaaaaagaa acctgttgcg
     2821 aaccgcgggg tgaatcccaa aaacgaatgc gttttggtgg agtgattgat tcgaatcgaa
     2881 gaagaaaaag aagaagacgt ggaatagaga gctcactctt aaccgagcag cacacaccga
     2941 cagaaaaaaa aatgaaatga atgagggtct tcttcttctt cttcttcgaa tgattgacag
     3001 aaatgggaaa aagaggaaga ttgagaaggg aaaaaggaag gagaaaagaa gcagaagaag
     3061 acgtcagaga ggagaggaac gagcggaaaa gcagcgggcg caagtcatag aagtagcaga
     3121 gctggggaga agaagacact atccaagaaa ggaatgacga gagagtatgc aaaggggtat
     3181 agggtgcaga cagaatagga acagaataac agatgatgag ccaagaagag ttgaaaaggg
     3241 cgatgaattt gtcatgtaac ttaatttggg tcaatttgag catgatgaat tgaaatcatc
     3301 ccttgttggg agttaataac cggtttgtta tcagaaaccc tgtaatagaa gggcgcccta
     3361 actttgagcc aattcatccc ggtttctgtc aaatatatca aaaagtggtc aactgacaaa
     3421 ttgtttttga tattataata aacattttat ccgttaacaa ttttcgaata ctttttacaa
     3481 ggacttggat aaattggctc aaagataact tcgtataatg tatgctatac gaagttatta
     3541 actaatctga tttaaatttt caggtgaaga caacttacaa agccaaaaaa ccagttcagc
     3601 tgccaggagc gtacaatgtt aatattaaac tggatatcac ctcccacaac gaggattaca
     3661 ctatcgttga gcaatatgaa agagctgaag ggcggcactc gacaggtggc atggatgaat
     3721 tgtataaggg tcgaagagca cccagctttc ttgtacaaag ttggcattat aagaaagcat
     3781 tgcttatcaa tttgttgcaa cgaacaggtc actatcagtc aaaataaaat cattatttgc
     3841 catccagctg atatccccta tagtgagtcg tattacatgg tcatagctgt ttcctggcag
     3901 ctctggcccg tgtctcaaaa tctctgatgt tacattgcac aagataaaat aatatcatca
     3961 tgaacaataa aactgtctgc ttacataaac agtaatacaa ggggtgttat gagccatatt
     4021 caacgggaaa cgtcgaggcc gcgattaaat tccaacatgg atgctgattt atatgggtat
     4081 aaatgggctc gcgataatgt cgggcaatca ggtgcgacaa tctatcgctt gtatgggaag
     4141 cccgatgcgc cagagttgtt tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca
     4201 gatgagatgg tcagactaaa ctggctgacg gaatttatgc ctcttccgac catcaagcat
     4261 tttatccgta ctcctgatga tgcatggtta ctcaccactg cgatccccgg aaaaacagca
     4321 ttccaggtat tagaagaata tcctgattca ggtgaaaata ttgttgatgc gctggcagtg
     4381 ttcctgcgcc ggttgcattc gattcctgtt tgtaattgtc cttttaacag cgatcgcgta
     4441 tttcgtctcg ctcaggcgca atcacgaatg aataacggtt tggttgatgc gagtgatttt
     4501 gatgacgagc gtaatggctg gcctgttgaa caagtctgga aagaaatgca taaacttttg
     4561 ccattctcac cggattcagt cgtcactcat ggtgatttct cacttgataa ccttattttt
     4621 gacgagggga aattaatagg ttgtattgat gttggacgag tcggaatcgc agaccgatac
     4681 caggatcttg ccatcctatg gaactgcctc ggtgagtttt ctccttcatt acagaaacgg
     4741 ctttttcaaa aatatggtat tgataatcct gatatgaata aattgcagtt tcatttgatg
     4801 ctcgatgagt ttttctaatc agaattggtt aattggttgt aacactggca gagcattacg
     4861 ctgacttgac gggacggcgc aagctcatga ccaaaatccc ttaacgtgag ttacgcgtcg
     4921 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     4981 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     5041 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     5101 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     5161 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     5221 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     5281 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     5341 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     5401 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     5461 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     5521 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     5581 ttcctggcct tttgctggcc ttttgctcac atgtt
//
