LOCUS       sequence-136624-        2918 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  SapTrap STOP codon + let-858 UTR N-terminal connector donor plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2918)
  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 2918)
  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: pMLS297
FEATURES             Location/Qualifiers
     source          1..2918
                     /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"
     primer_bind     563..582
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        563..581
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     606..622
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     607..623
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1091..1107)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1091..1107)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(1144..1164)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1144..1162)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1183..1199)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1183..1199)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1301..1320)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1312..2121
                     /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(1379..1398)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2268..2856
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2757..2776
                     /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 agtgagcgcg cgtaatacga ctcactatag ggcgaattgg gtaccgggcc
      601 ccccctcgag gtcgacggta tcgataagct tgatatcgaa ttcctgcagc ccggctcttc
      661 gggttaaatt ttcaaatttt aaatactgaa tatttgtttt ttttcctatt atttatttat
      721 tctctttgtg ttttttttct tgctttctaa aaaattaatt caatccaaat ctaaacattt
      781 ttttttctct ttccgtctcc caattcgtat tccgctcctc tcatctgaac acaatgtgca
      841 agtttattta tcttctcgct ttcatttcat taggacgtgg ggggaattgg tggaaggggg
      901 aaacacacaa aaggatgatg gaaatgaaat aaggacacac aatatgcaac aacattcaat
      961 tcagaaatat ggaggaaggt ttaaaagaaa acataaaaat atatagagga ggaaggaaaa
     1021 ctagtaaaaa ataagcaaag aaattaggcg aacgatgaga attgtcctcg cttgacggga
     1081 agagcggggg gatccactag ttctagagcg gccgccaccg cggtggagct ccagcttttg
     1141 ttccctttag tgagggttaa ttgcgcgctt ggcgtaatca tggtcatagc tgtttcctgg
     1201 cagctctggc ccgtgtctca aaatctctga tgttacattg cacaagataa aaatatatca
     1261 tcatgaacaa taaaactgtc tgcttacata aacagtaata caaggggtgt tatgagccat
     1321 attcaacggg aaacgtcgag gccgcgatta aattccaaca tggatgctga tttatatggg
     1381 tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga caatctatcg cttgtatggg
     1441 aagcccgatg cgccagagtt gtttctgaaa catggcaaag gtagcgttgc caatgatgtt
     1501 acagatgaga tggtcagact aaactggctg acggaattta tgcctcttcc gaccatcaag
     1561 cattttatcc gtactcctga tgatgcatgg ttactcacca ctgcgatccc cggaaaaaca
     1621 gcattccagg tattagaaga atatcctgat tcaggtgaaa atattgttga tgcgctggca
     1681 gtgttcctgc gccggttgca ttcgattcct gtttgtaatt gtccttttaa cagcgatcgc
     1741 gtatttcgtc tcgctcaggc gcaatcacga atgaataacg gtttggttga tgcgagtgat
     1801 tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct ggaaagaaat gcataaactt
     1861 ttgccattct caccggattc agtcgtcact catggtgatt tctcacttga taaccttatt
     1921 tttgacgagg ggaaattaat aggttgtatt gatgttggac gagtcggaat cgcagaccga
     1981 taccaggatc ttgccatcct atggaactgc ctcggtgagt tttctccttc attacagaaa
     2041 cggctttttc aaaaatatgg tattgataat cctgatatga ataaattgca gtttcatttg
     2101 atgctcgatg agtttttcta atcagaattg gttaattggt tgtaacactg gcagagcatt
     2161 acgctgactt gacgggacgg cgcaagctca tgaccaaaat cccttaacgt gagttacgcg
     2221 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     2281 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     2341 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     2401 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     2461 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     2521 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     2581 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     2641 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     2701 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     2761 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     2821 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     2881 cggttcctgg ccttttgctg gccttttgct cacatgtt
//
