LOCUS       sequence-345252-        2494 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  htz-1 targeting gRNA expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2494)
  AUTHORS   Negishi T, Kitagawa S, Horii N, Tanaka Y, Haruta N, Sugimoto A, Sawa
            H, Hayashi KI, Harata M, Kanemaki MT
  TITLE     The auxin-inducible degron 2 (AID2) system enables controlled 
            protein knockdown during embryogenesis and development in 
            Caenorhabditis elegans.
  JOURNAL   Genetics. 2021 Dec 2. pii: 6448232. doi: 10.1093/genetics/iyab218.
  PUBMED    34865044
REFERENCE   2  (bases 1 to 2494)
  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. 
            2021 Dec 2. pii: 6448232. doi: 10.1093/genetics/iyab218."
COMMENT     Sequence Label: pTK73_htz-1
FEATURES             Location/Qualifiers
     source          1..2494
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(234..250)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(234..250)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     terminator      298..329
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        330..432
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             433..1248
                     /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     complement(506..525)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      1272..1299
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(1311..1898)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1391..1410)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1938..1955
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1939..1955
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2130..2479
                     /label=CeU6 promoter
                     /note="promoter for the C. elegans R07E5.16 U6 snRNA gene 
                     (Dickinson et al., 2013)"
ORIGIN
        1 cgcaagttta agagctatgc tggaaacagc atagcaagtt taaataaggc tagtccgtta
       61 tcaacttgaa aaagtggcac cgagtcggtg ctttttgtga aatttgctag cggtgcgagc
      121 ggatcgagca gtgtcgatca ctactggacc gcgagctgtg ctgcgacccg tgatcttacg
      181 gcattatacg tatgatcggt ccacgatcag ctagattatc tagtcagctt gatgtcatag
      241 ctgtttcctg aggctcaata ctgaccattt aaatcatacc tgacctccat agcagaaagt
      301 caaaagcctc cgaccggagg cttttgactt gatcggcacg taagaggttc caactttcac
      361 cataatgaaa taagatcact accgggcgta ttttttgagt tatcgagatt ttcaggagct
      421 aaggaagcta aaatgagcca tattcaacgg gaaacgtctt gcttgaagcc gcgattaaat
      481 tccaacatgg atgctgattt atatgggtat aaatgggctc gcgataatgt cgggcaatca
      541 ggtgcgacaa tctatcgatt gtatgggaag cccgatgcgc cagagttgtt tctgaaacat
      601 ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg tcaggctaaa ctggctgacg
      661 gaatttatgc ctcttccgac catcaagcat tttatccgta ctcctgatga tgcatggtta
      721 ctcaccactg cgatcccagg gaaaacagca ttccaggtat tagaagaata tcctgattca
      781 ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc ggttgcattc gattcctgtt
      841 tgtaattgtc cttttaacgg cgatcgcgta tttcgtctcg ctcaggcgca atcacgaatg
      901 aataacggtt tggttggtgc gagtgatttt gatgacgagc gtaatggctg gcctgttgaa
      961 caagtctgga aagaaatgca taaactcttg ccattctcac cggattcagt cgtcactcat
     1021 ggtgatttct cacttgataa ccttattttt gacgagggga aattaatagg ttgtattgat
     1081 gttggacgag tcggaatcgc agaccgatac caggatcttg ccatcctatg gaactgcctc
     1141 ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa aatatggtat tgataatcct
     1201 gatatgaata aattgcagtt tcacttgatg ctcgatgagt ttttctaatg aggacctaaa
     1261 tgtaatcacc tggctcacct tcgggtgggc ctttctgcgt tgctggcgtt tttccatagg
     1321 ctccgccccc ctgacgagca tcacaaaaat cgatgctcaa gtcagaggtg gcgaaacccg
     1381 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     1441 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     1501 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     1561 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     1621 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     1681 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     1741 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac ctcggaaaaa
     1801 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     1861 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg attttctacc
     1921 gaagaaaggc ccacccgtgt aaaacgacgg ccagtttatc tagtcagctt gattctagct
     1981 gatcgtggac cggaaggtga gccagtgagt tgattgcagt ccagttacgc tggagtctga
     2041 ggctcgtcct gaatgatatg cgaccgccgg agggttgcgt ttgagacggg cgacagatcc
     2101 agtcgcgctg ctctcgtcga tccaagcttc aaaaaaaact agcaataaag gaataaaaaa
     2161 ctgtacacct taaaggcgca cactctgttt tgcaaatttt atttttagtt gtgaattttc
     2221 tgctgagact tgaaaatagc aactttagta ctactataat ttgtcaacct tttcaaaaaa
     2281 agcatgcaat ttttgagaaa ctcttataaa agctattatt aaaaaaacac cttttttcca
     2341 aaattattcc acaaaaaata tgttatgaaa tgcctacacc ctctcacaca cactctttat
     2401 actactctgt caaactcacg agatgtctgc cgcctcttgt gttgccccta tataaacacc
     2461 tcctattgcg agatgtctta gcttcaatct agtt
//
