LOCUS       sequence-136604-        2860 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  SapTrap SL2::SV40-NLS C-terminal connector donor plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2860)
  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 2860)
  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: pMLS269
FEATURES             Location/Qualifiers
     source          1..2860
                     /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"
     CDS             940..960
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     protein_bind    complement(985..1084)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1101..1120)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1102..1120)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1125..1141)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1125..1141)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1243..1262)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1254..2063
                     /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(1321..1340)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2210..2798
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2699..2718
                     /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 agcaggctat gctcttcggc gtaagctgtc tcatcctact ttcacctagt taactgcttg
      721 tcttaaaatc tatgcttctc tttagtatct aaaattttcc tagaagctta caagtatata
      781 aatggtctct tctcaataaa ggttgtatat ttattcatct tattgaatct gccatttcct
      841 cgtttttgcg agtttatata ccttccaatt ttctttctat tgtattttca acttctaatt
      901 ttaattcagg gaaactgctt caacgcatca atgactgctc caaagaagaa gcgtaaggta
      961 ccggtagaaa aaatgcgaag agctacccag ctttcttgta caaagttggc attataagaa
     1021 agcattgctt atcaatttgt tgcaacgaac aggtcactat cagtcaaaat aaaatcatta
     1081 tttgccatcc agctgatatc ccctatagtg agtcgtatta catggtcata gctgtttcct
     1141 ggcagctctg gcccgtgtct caaaatctct gatgttacat tgcacaagat aaaataatat
     1201 catcatgaac aataaaactg tctgcttaca taaacagtaa tacaaggggt gttatgagcc
     1261 atattcaacg ggaaacgtcg aggccgcgat taaattccaa catggatgct gatttatatg
     1321 ggtataaatg ggctcgcgat aatgtcgggc aatcaggtgc gacaatctat cgcttgtatg
     1381 ggaagcccga tgcgccagag ttgtttctga aacatggcaa aggtagcgtt gccaatgatg
     1441 ttacagatga gatggtcaga ctaaactggc tgacggaatt tatgcctctt ccgaccatca
     1501 agcattttat ccgtactcct gatgatgcat ggttactcac cactgcgatc cccggaaaaa
     1561 cagcattcca ggtattagaa gaatatcctg attcaggtga aaatattgtt gatgcgctgg
     1621 cagtgttcct gcgccggttg cattcgattc ctgtttgtaa ttgtcctttt aacagcgatc
     1681 gcgtatttcg tctcgctcag gcgcaatcac gaatgaataa cggtttggtt gatgcgagtg
     1741 attttgatga cgagcgtaat ggctggcctg ttgaacaagt ctggaaagaa atgcataaac
     1801 ttttgccatt ctcaccggat tcagtcgtca ctcatggtga tttctcactt gataacctta
     1861 tttttgacga ggggaaatta ataggttgta ttgatgttgg acgagtcgga atcgcagacc
     1921 gataccagga tcttgccatc ctatggaact gcctcggtga gttttctcct tcattacaga
     1981 aacggctttt tcaaaaatat ggtattgata atcctgatat gaataaattg cagtttcatt
     2041 tgatgctcga tgagtttttc taatcagaat tggttaattg gttgtaacac tggcagagca
     2101 ttacgctgac ttgacgggac ggcgcaagct catgaccaaa atcccttaac gtgagttacg
     2161 cgtcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     2221 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     2281 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     2341 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     2401 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     2461 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     2521 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     2581 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     2641 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     2701 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     2761 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     2821 tacggttcct ggccttttgc tggccttttg ctcacatgtt
//
