LOCUS       sequence-136506-        6541 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  SapTrap 3-site Destination vector for N-terminal GFP tagging with 
            embedded Cbr-unc-119 selectable marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6541)
  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 6541)
  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: pMLS234
FEATURES             Location/Qualifiers
     source          1..6541
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     117..134
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        141..639
                     /label=CeU6 promoter
                     /note="promoter for the C. elegans K09B11.12 U6 snRNA gene 
                     (Friedland et al., 2013)"
     misc_RNA        667..742
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    750..986
                     /label=3' CeU6
                     /note="region downstream of the C. elegans K09B11.12 U6 
                     snRNA gene"
     terminator      complement(1132..1159)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(1251..1337)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     1386..1408
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1400..1417
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1401..1417
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1433..1532
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1594..1622)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     protein_bind    2181..2214
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             4178..4189
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     protein_bind    4338..4371
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     4561..4581
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     protein_bind    complement(4691..4790)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(4807..4826)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4808..4826)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4831..4847)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4831..4847)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4949..4968)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4960..5769
                     /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(5027..5046)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      5916..6504
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6405..6424
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
       61 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
      121 agtcagtgag cgaggaagcg tccaagaact cgtacaaaaa tgctctgaag taggtctcga
      181 gatcaatact gggaagacga aagtcttgcg aaaccgattc gctgacccca gtaaagtcta
      241 cttcggtagc ccttccccca ccacccagct cgacgacgtc gacgagtaca tctacctcgg
      301 tcgtcaaatc aacgcccaaa acaacttgat gccggaaatc caccgaagac gtcgagcagc
      361 ctgggctgca ttcaatggaa tcaagaatgc caccgactcc atcaccgaca agaagattcg
      421 tgcgaatctg ttcgactcaa ttgtccttcc agcgctcacc tacgtttcag aagcctggac
      481 attcaccaaa gctctatccg aacgagtacg aatcacacat gcctccccca caaattattg
      541 atctacatcc ccatttggta tagtgtctta caggactctc aatatgtcag ctgcatgata
      601 aatggtccct atataattga attgcaaatc taaatgtttg cgaagagcgg atccatgggc
      661 tcttcggttt tagagctaga aatagcaagt taaaataagg ctagtccgtt atcaacttga
      721 aaaagtggca ccgagtcggt gctttttttc aataatattc tagatatccc tttttgaaac
      781 taaaatgaaa tactctaaat ttttgagttc aatgtttcaa ctaatcttgg ttaaaggtaa
      841 attcccggag ggacgatttt aaagttgcca gagttgatta aattggtcca aaaaatttta
      901 ttaggaaact cgaaaaaaaa ttgatttttt ttgacaatat tccaaagaaa attacaaaat
      961 ctcgacgcca aacatagtcg gctgtggccc aatacgcaaa ccgcctctcc ccgcgcgttg
     1021 gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg
     1081 caacgcaatt aatacgcgta ccgctagcca ggaagagttt gtagaaacgc aaaaaggcca
     1141 tccgtcagga tggccttctg cttagtttga tgcctggcag tttatggcgg gcgtcctgcc
     1201 cgccaccctc cgggccgttg cttcacaacg ttcaaatccg ctcccggcgg atttgtccta
     1261 ctcaggagag cgttcaccga caaacaacag ataaaacgaa aggcccagtc ttccgactga
     1321 gcctttcgtt ttatttgatg cctggcagtt ccctactctc gcgttaacgc tagcatggat
     1381 gttttcccag tcacgacgtt gtaaaacgac ggccagtctt aagctcgggc cccaaataat
     1441 gattttattt tgactgatag tgacctgttc gttgcaacaa attgatgagc aatgcttttt
     1501 tataatgcca actttgtaca aaaaagcagg ctcgtaccga agagcggatc catgggctct
     1561 tcgatgagca aaggagaaga acttttcact ggagttgtcc caattcttgt tgaattagat
     1621 ggtgatgtta atgggcacaa attttctgtc agtggagagg gtgaaggtga tgcaacatac
     1681 ggaaaactta cccttaaatt tatttgcact actggaaaac tacctgttcc atgggtaagt
     1741 ttaaacatat atatactaac taaccctgat tatttaaatt ttcagccaac acttgtcact
     1801 actttctgtt atggtgttca atgcttctcg agatacccag atcatatgaa acggcatgac
     1861 tttttcaaga gtgccatgcc cgaaggttat gtacaggaaa gaactatatt tttcaaagat
     1921 gacgggaact acaagacacg taagtttaaa cagttcggta ctaactaacc atacatattt
     1981 aaattttcag gtgctgaagt caagtttgaa ggtgataccc ttgttaatag aatcgagtta
     2041 aaaggtattg attttaaaga agatggaaac attcttggac acaaattgga atacaactat
     2101 aactcacaca atgtatacat catggcagac aaacaaaaga atggaatcaa agttgtaagt
     2161 ttaaacatga ttttactaac ataacttcgt ataatgtatg ctatacgaag ttatttctag
     2221 acattctcta atgaaaaaat ctttcagttg aaattgaaaa tgagttaaag ttggagtttt
     2281 tattgaaaac agatttccgt gtgattagtg tttttagcga gtgtgacagg acagcgaaaa
     2341 aatatagaaa caagggggga actgaaaagc ttaggaatgc attgaacatg agaaggggaa
     2401 ggggaaggaa caaactagac aggaattatt ggaatttaat cacatttgga gttttttttc
     2461 tattcgacag aataattatc cagaacattt ttgtattaaa tatttatgca tcatatgagt
     2521 agtcggcttt gttgtgcatg acgagtttgt tatcgacgaa atagaagctg tcagaacgag
     2581 tctcgtttgg attgttgatc atgtcgtcca ctgaaaaaga gattagtctt ttgaattgta
     2641 ctttttagaa taatgactca ctgagttgtt gagagagttg agggaactca tagatatgtt
     2701 cacagttgtt tcgtgaattc ggaatacaga atccgaattc aaagtcaaaa cacttcagaa
     2761 ggcgatcttt gaagaagtga cgttcgatca ttcggaaatg atggattggg atgtctccta
     2821 ctttgaattc cacagttgct ccgaccgttt tgagtttcaa aaagtttgga gcaaatctat
     2881 aacgcacgta tcttgccgac tcctgtggcg actcatcatt ctcctgatcg ttctccggtt
     2941 tggcgatctc gaaaagcact tgctcagtgt ccagatcacg gatttggaac ttggtgaact
     3001 cgatgttata gatgttcgca gatggggagc ataagaatcc taaatttatg ttttaaactg
     3061 aaatccaaag ggagcaagat accttgagtg attcccggaa gtgctaaaac gtcgttcgga
     3121 gtgatttgag ctttcttcgc aagctccgat tccgttgtga ttccttgttc ggtgcttggt
     3181 ggtggccgtg gcatctggaa atatggaaaa gttcaacaaa aagaaaagag aaaagaatga
     3241 aatcggatat caagagttag ttgagcggtt tctctagttt tctgagtctc acctgcgacg
     3301 ggaaggtcgc cgagccgggt ggaatcgatt gttgttgctc ggctttcata tcggtttggt
     3361 tggaagcggc tgaaaacgga aagaagtgga agaaggaaaa gagtgtggtg tgacaggaaa
     3421 atggtaatta gagggtgcca aataaccagc tatattttgt ttttttttga aaacattttt
     3481 aaaaagaaaa atacgataat gatatcagat ggatttccgg aaaactggta tgaaaaattt
     3541 caaccttttt gagtacatgt aatcaaaata cactttgtaa attatcattt ttattgaaac
     3601 tccaccattt ttctatttat aacgctaata atttgaaaaa gaaacctgtt gcgaaccgcg
     3661 gggtgaatcc caaaaacgaa tgcgttttgg tggagtgatt gattcgaatc gaagaagaaa
     3721 aagaagaaga cgtggaatag agagctcact cttaaccgag cagcacacac cgacagaaaa
     3781 aaaaatgaaa tgaatgaggg tcttcttctt cttcttcttc gaatgattga cagaaatggg
     3841 aaaaagagga agattgagaa gggaaaaagg aaggagaaaa gaagcagaag aagacgtcag
     3901 agaggagagg aacgagcgga aaagcagcgg gcgcaagtca tagaagtagc agagctgggg
     3961 agaagaagac actatccaag aaaggaatga cgagagagta tgcaaagggg tatagggtgc
     4021 agacagaata ggaacagaat aacagatgat gagccaagaa gagttgaaaa gggcgatgaa
     4081 tttgtcatgt aacttaattt gggtcaattt gagcatgatg aattgaaatc atcccttgtt
     4141 gggagttaat aaccggtttg ttatcagaaa ccctgtaata gaagggcgcc ctaactttga
     4201 gccaattcat cccggtttct gtcaaatata tcaaaaagtg gtcaactgac aaattgtttt
     4261 tgatattata ataaacattt tatccgttaa caattttcga atacttttta caaggacttg
     4321 gataaattgg ctcaaagata acttcgtata atgtatgcta tacgaagtta ttaactaatc
     4381 tgatttaaat tttcagaact tcaaaattag acacaacatt gaagatggaa gcgttcaact
     4441 agcagaccat tatcaacaaa atactccaat tggcgatggc cctgtccttt taccagacaa
     4501 ccattacctg tccacacaat ctgccctttc gaaagatccc aacgaaaaga gagaccacat
     4561 ggtccttctt gagtttgtaa cagctgctgg gattacacat ggcatggatg aactatacaa
     4621 aggtggcaca ggaggaacgg gcggtagtgg aggcactggt cgaagagcgg atccatgggc
     4681 tcttcgccag acccagcttt cttgtacaaa gttggcatta taagaaagca ttgcttatca
     4741 atttgttgca acgaacaggt cactatcagt caaaataaaa tcattatttg ccatccagct
     4801 gatatcccct atagtgagtc gtattacatg gtcatagctg tttcctggca gctctggccc
     4861 gtgtctcaaa atctctgatg ttacattgca caagataaaa taatatcatc atgaacaata
     4921 aaactgtctg cttacataaa cagtaataca aggggtgtta tgagccatat tcaacgggaa
     4981 acgtcgaggc cgcgattaaa ttccaacatg gatgctgatt tatatgggta taaatgggct
     5041 cgcgataatg tcgggcaatc aggtgcgaca atctatcgct tgtatgggaa gcccgatgcg
     5101 ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca atgatgttac agatgagatg
     5161 gtcagactaa actggctgac ggaatttatg cctcttccga ccatcaagca ttttatccgt
     5221 actcctgatg atgcatggtt actcaccact gcgatccccg gaaaaacagc attccaggta
     5281 ttagaagaat atcctgattc aggtgaaaat attgttgatg cgctggcagt gttcctgcgc
     5341 cggttgcatt cgattcctgt ttgtaattgt ccttttaaca gcgatcgcgt atttcgtctc
     5401 gctcaggcgc aatcacgaat gaataacggt ttggttgatg cgagtgattt tgatgacgag
     5461 cgtaatggct ggcctgttga acaagtctgg aaagaaatgc ataaactttt gccattctca
     5521 ccggattcag tcgtcactca tggtgatttc tcacttgata accttatttt tgacgagggg
     5581 aaattaatag gttgtattga tgttggacga gtcggaatcg cagaccgata ccaggatctt
     5641 gccatcctat ggaactgcct cggtgagttt tctccttcat tacagaaacg gctttttcaa
     5701 aaatatggta ttgataatcc tgatatgaat aaattgcagt ttcatttgat gctcgatgag
     5761 tttttctaat cagaattggt taattggttg taacactggc agagcattac gctgacttga
     5821 cgggacggcg caagctcatg accaaaatcc cttaacgtga gttacgcgtc gttccactga
     5881 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     5941 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     6001 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     6061 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     6121 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     6181 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     6241 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     6301 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     6361 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     6421 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     6481 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     6541 t
//
