LOCUS       sequence-460827-        1928 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Entry vector with reverse FRT3 (rFRT3) in slot 7 (for SapTrap 
            assembly).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 1928)
  AUTHORS   Khan H, Huang X, Raj V, Wang H
  TITLE     A versatile site-directed gene trap strategy to manipulate gene 
            activity and control gene expression in Caenorhabditis elegans.
  JOURNAL   PLoS Genet. 2025 Jan 22;21(1):e1011541. doi: 
            10.1371/journal.pgen.1011541. eCollection 2025 Jan.
  PUBMED    39841730
REFERENCE   2  (bases 1 to 1928)
  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: "PLoS 
            Genet."; date: "2025-01-22"; volume: "21(1):e1011541. doi"; pages: "
            10.1371/journal.pgen.1011541. eCollection 2025 Jan"
COMMENT     Sequence Label: pHW622
FEATURES             Location/Qualifiers
     source          1..1928
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(1..816)
                     /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="MSHIQRETSCLKPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNGDRVFRLAQAQSRMNNGLVGA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     724..743
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(817..919)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(920..951)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     protein_bind    complement(1003..1036)
                     /label=FRT3
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision with other FRT3 sites
                     (Bosch et al., 2015)"
     primer_bind     complement(1222..1239)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1222..1238)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      1279..1866
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1767..1786
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1878..1905)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
ORIGIN
        1 ttagaaaaac tcatcgagca tcaagtgaaa ctgcaattta ttcatatcag gattatcaat
       61 accatatttt tgaaaaagcc gtttctgtaa tgaaggagaa aactcaccga ggcagttcca
      121 taggatggca agatcctggt atcggtctgc gattccgact cgtccaacat caatacaacc
      181 tattaatttc ccctcgtcaa aaataaggtt atcaagtgag aaatcaccat gagtgacgac
      241 tgaatccggt gagaatggca agagtttatg catttctttc cagacttgtt caacaggcca
      301 gccattacgc tcgtcatcaa aatcactcgc accaaccaaa ccgttattca ttcgtgattg
      361 cgcctgagcg agacgaaata cgcgatcgcc gttaaaagga caattacaaa caggaatcga
      421 atgcaaccgg cgcaggaaca ctgccagcgc atcaacaata ttttcacctg aatcaggata
      481 ttcttctaat acctggaatg ctgttttccc tgggatcgca gtggtgagta accatgcatc
      541 atcaggagta cggataaaat gcttgatggt cggaagaggc ataaattccg tcagccagtt
      601 tagcctgacc atctcatctg taacatcatt ggcaacgcta cctttgccat gtttcagaaa
      661 caactctggc gcatcgggct tcccatacaa tcgatagatt gtcgcacctg attgcccgac
      721 attatcgcga gcccatttat acccatataa atcagcatcc atgttggaat ttaatcgcgg
      781 cttcaagcaa gacgtttccc gttgaatatg gctcatttta gcttccttag ctcctgaaaa
      841 tctcgataac tcaaaaaata cgcccggtag tgatcttatt tcattatggt gaaagttgga
      901 acctcttacg tgccgatcaa gtcaaaagcc tccggtcgga ggcttttgac tttctgctat
      961 ggaggtcagg tatgattttt gaattcgctc ttcgggtccc gggaagttcc tatactattt
     1021 gaagaatagg aacttcacgc gaagagcaag cttggatcga cgagagcagc gcgactggat
     1081 ctgtcgcccg tctcaaacgc aaccctccgg cggtcgcata tcattcagga cgagcctcag
     1141 actccagcgt aactggactg caatcaactc actggctcac cttccggtcc acgatcagct
     1201 agaatcaagc tgactagata aactggccgt cgttttacac gggtgggcct ttcttcggta
     1261 gaaaatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     1321 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     1381 tccgaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     1441 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     1501 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     1561 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     1621 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     1681 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     1741 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     1801 tttcgccacc tctgacttga gcatcgattt ttgtgatgct cgtcaggggg gcggagccta
     1861 tggaaaaacg ccagcaacgc agaaaggccc acccgaaggt gagccaggtg attacattta
     1921 ggtcctca
//
