LOCUS       sequence-136628-        4016 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway compatible [1-2] Entry Vector encoding 2xNLS-FLP-D5.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4016)
  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 4016)
  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: pWD509
FEATURES             Location/Qualifiers
     source          1..4016
                     /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             677..697
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2066..2137
                     /codon_start=1
                     /product="nuclear localization signal from the C. elegans 
                     EGL-13 transcription factor (Lyssenko et al., 2007)"
                     /label=EGL-13 NLS
                     /translation="SRRRKANPTKLSENAKKLAKEVEN"
     protein_bind    complement(2141..2240)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2257..2276)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2258..2276)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2281..2297)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2281..2297)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2399..2418)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2410..3219
                     /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(2477..2496)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3366..3954
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3855..3874
                     /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 agcaggctaa aaaatgccaa agaagaagcg taaggtccca caatttgaca tcctttgtaa
      721 aaccccacca aaggtcctcg tccgtcaatt cgtcgagcgt ttcgagcgtc catccggaga
      781 gaagatcgcc ctctgcgccg ccgagctcac ctacctctgc tggatgatca cccacaacgg
      841 aaccgccatc aagcgtgcca ccttcatgtc ctacaacacc atcatctcca actccctctc
      901 cttcgacatc gtcaacaagt ccctccaatt caagtacaag acccaaaagg ccaccatcct
      961 cgaggcctcc ctcaagaagc tcatcccagc ctgggagttc accatcatcc catactacgg
     1021 acaaaagcac caatccgaca tcaccgacat cgtctcctcc ctccaactcc aattcgagtc
     1081 ctccgaggag gccgacaagg gaaactccca ctccaagaag gtaagtttaa acatatatat
     1141 actaactaac cctgattatt taaattttca gatgctcaag gccctcctct ccgagggaga
     1201 gtccatctgg gagatcaccg agaagatcct caactccttc gagtacacct cccgtttcac
     1261 caagaccaag accctctacc aattcctctt cctcgccacc ttcatcaact gcggacgttt
     1321 ctccgacatc aagaacgtcg acccaaagtc cttcaagctc gtccaaaaca agtacctcgg
     1381 agtcatcatc caatgcctcg tcaccgagac caagacctcc gtctcccgtc acatctactt
     1441 cttctccgcc cgtggacgta tcgacccact cgtctacctc gacgagttcc tccgtaactc
     1501 cgagccagtc ctcaagcgtg tcaaccgtac cggaaactcc tcctccaaca agcaagagta
     1561 ccaactcctc aaggacaacc tcgtccgttc ctacaacaag gtaagtttaa acagttcggt
     1621 actaactaac catacatatt taaattttca ggccctcaag aagaacgccc catactccat
     1681 cttcgccatc aagaacggac caaagtccca catcggacgt cacctcatga cctccttcct
     1741 ctccatgaag ggactcaccg agctcaccaa cgtcgtcgga aactggtccg acaagcgtgc
     1801 ctccgccgtc gcccgtacca cctacaccca ccaaatcacc gccatcccag accactactt
     1861 cgccctcgtc tcccgttact acgcctacga cccaatctcc aaggagatga tcgccctcaa
     1921 ggacgagacc aacccaatcg aggagtggca acacatcgag caactcaagg gttccgccga
     1981 gggatccatc cgttacccag cctggaacgg aatcatctcc caagaggtcc tcgactacct
     2041 ctcctcctac atcaaccgtc gtatctcccg tcgtcgtaag gccaacccaa ccaagctctc
     2101 cgagaacgcc aagaagctcg ccaaggaggt cgagaactaa acccagcttt cttgtacaaa
     2161 gttggcatta taagaaagca ttgcttatca atttgttgca acgaacaggt cactatcagt
     2221 caaaataaaa tcattatttg ccatccagct gatatcccct atagtgagtc gtattacatg
     2281 gtcatagctg tttcctggca gctctggccc gtgtctcaaa atctctgatg ttacattgca
     2341 caagataaaa taatatcatc atgaacaata aaactgtctg cttacataaa cagtaataca
     2401 aggggtgtta tgagccatat tcaacgggaa acgtcgaggc cgcgattaaa ttccaacatg
     2461 gatgctgatt tatatgggta taaatgggct cgcgataatg tcgggcaatc aggtgcgaca
     2521 atctatcgct tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca tggcaaaggt
     2581 agcgttgcca atgatgttac agatgagatg gtcagactaa actggctgac ggaatttatg
     2641 cctcttccga ccatcaagca ttttatccgt actcctgatg atgcatggtt actcaccact
     2701 gcgatccccg gaaaaacagc attccaggta ttagaagaat atcctgattc aggtgaaaat
     2761 attgttgatg cgctggcagt gttcctgcgc cggttgcatt cgattcctgt ttgtaattgt
     2821 ccttttaaca gcgatcgcgt atttcgtctc gctcaggcgc aatcacgaat gaataacggt
     2881 ttggttgatg cgagtgattt tgatgacgag cgtaatggct ggcctgttga acaagtctgg
     2941 aaagaaatgc ataaactttt gccattctca ccggattcag tcgtcactca tggtgatttc
     3001 tcacttgata accttatttt tgacgagggg aaattaatag gttgtattga tgttggacga
     3061 gtcggaatcg cagaccgata ccaggatctt gccatcctat ggaactgcct cggtgagttt
     3121 tctccttcat tacagaaacg gctttttcaa aaatatggta ttgataatcc tgatatgaat
     3181 aaattgcagt ttcatttgat gctcgatgag tttttctaat cagaattggt taattggttg
     3241 taacactggc agagcattac gctgacttga cgggacggcg caagctcatg accaaaatcc
     3301 cttaacgtga gttacgcgtc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     3361 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     3421 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     3481 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     3541 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     3601 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     3661 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     3721 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     3781 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     3841 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     3901 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     3961 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgtt
//
