LOCUS       sequence-424143-        3389 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express KnFnuc guide with pureexpress.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3389)
  AUTHORS   Jiang K, Lim J, Sgrizzi S, Trinh M, Kayabolen A, Yutin N, Bao W, 
            Kato K, Koonin EV, Gootenberg JS, Abudayyeh OO
  TITLE     Programmable RNA-guided DNA endonucleases are widespread in 
            eukaryotes and their viruses.
  JOURNAL   Sci Adv. 2023 Sep 29;9(39):eadk0171. doi: 10.1126/sciadv.adk0171. 
            Epub 2023 Sep 27.
  PUBMED    37756409
REFERENCE   2  (bases 1 to 3389)
  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: "Sci Adv."; 
            date: "2023-09-29"; volume: "9(39):eadk0171. doi"; pages: " 
            10.1126/sciadv.adk0171. Epub 2023 Sep 27"
COMMENT     Sequence Label: pKJ238
FEATURES             Location/Qualifiers
     source          1..3389
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     ncRNA           201..268
                     /product="hepatitis delta virus self-cleaving ribozyme 
                     (Ferre-D'Amare and Scott, 2010)"
                     /label=HDV ribozyme
                     /ncRNA_class="ribozyme"
     terminator      277..324
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(649..668)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             660..1475
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             660..1475
                     /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="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(733..752)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1592..1609
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(1633..1690)
                     /label=his operon terminator
                     /note="This putative transcriptin terminator from the E. 
                     coli his operon has a 2-bp deletion introduced during 
                     synthesis. Its efficiency has not been determined."
     rep_origin      2181..2769
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2670..2689
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3218..3237
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3218..3236
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    3236..3260
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
ORIGIN
        1 gcatagtcag aggcgagggc aggccagcgg agttcacgag agcaggaatg tgaggatgac
       61 tgagaattag tcgaaagaca tagctgccta gaaacgagtt catctaggca cttcggtgag
      121 aatccgagat acggctgggt actgtggcga gtgtgccatt ttactctgaa cagactgtag
      181 cagccacctc cttgttattg ggccggcatg gtcccagcct cctcgctggc gccggctggg
      241 caacatgctt cggcatggcg aatgggacgc ggccgcctag cataacccct tggggcctct
      301 aaacgggtct tgaggggttt tttggctagc gctgaggtct gcctcgtgaa gaaggtgttg
      361 ctgactcata ccaggcctga atcgccccat catccagcca gaaagtgagg gagccacggt
      421 tgatgagagc tttgttgtag gtggaccagt tggtgatttt gaacttttgc tttgccacgg
      481 aacggtctgc gttgtcggga agatgcgtga tctgatcctt caactcagca aaagttcgat
      541 ttattcaaca aagccacgtt gtgtctcaaa atctctgatg ttacattgca caagataaaa
      601 atatatcatc atgaacaata aaactgtctg cttacataaa cagtaataca aggggtgtta
      661 tgagccatat tcaacgggaa acgtcttgct ccaggccgcg attaaattcc aacatggatg
      721 ctgatttata tgggtataaa tgggctcgcg ataatgtcgg gcaatcaggt gcgacaatct
      781 atcgattgta tgggaagccc gatgcgccag agttgtttct gaaacatggc aaaggtagcg
      841 ttgccaatga tgttacagat gagatggtca gactaaactg gctgacggaa tttatgcctc
      901 ttccgaccat caagcatttt atccgtactc ctgatgatgc atggttactc accactgcga
      961 tccccgggaa aacagcattc caggtattag aagaatatcc tgattcaggt gaaaatattg
     1021 ttgatgcgct ggcagtgttc ctgcgccggt tgcattcgat tcctgtttgt aattgtcctt
     1081 ttaacagcga tcgcgtattt cgtctcgctc aggcgcaatc acgaatgaat aacggtttgg
     1141 ttgatgcgag tgattttgat gacgagcgta atggctggcc tgttgaacaa gtctggaaag
     1201 aaatgcataa gcttttgcca ttctcaccgg attcagtcgt cactcatggt gatttctcac
     1261 ttgataacct tatttttgac gaggggaaat taataggttg tattgatgtt ggacgagtcg
     1321 gaatcgcaga ccgataccag gatcttgcca tcctatggaa ctgcctcggt gagttttctc
     1381 cttcattaca gaaacggctt tttcaaaaat atggtattga taatcctgat atgaataaat
     1441 tgcagtttca tttgatgctc gatgagtttt tctaatcaga attggttaat tggttgtaac
     1501 actggcagag cattacgctg acttgacggg actcgagatt accgcctttg agtgagctga
     1561 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcctgcat
     1621 aacgcgaagt aatcttttcg gttttaaaga aaaagggcag ggtggtgaca ccttgccctt
     1681 ttttgccgga ctgcagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     1741 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     1801 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     1861 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     1921 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     1981 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     2041 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     2101 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     2161 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     2221 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     2281 tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag
     2341 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     2401 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     2461 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     2521 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     2581 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     2641 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     2701 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     2761 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     2821 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     2881 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgaggt cggcgatata
     2941 ggcgcgagca accgcccctg tggcgccggt gatgccggcc acgatgcgtc cggcgtagag
     3001 gatctgctca tgtttgacag cttatcatcg atgcataatg tgcctgtcaa atggacgaag
     3061 cagggattct gcaaacccta tgctactccc tcgagccgtc aattgtctga ttcgttacca
     3121 attatactag ctccggcaaa aaaacgggca aggtgtcacc accctgccct ttttctttaa
     3181 aaccgaaaag attacttcgc gtttgccacc tgacgtctaa tacgactcac tatagggaat
     3241 tgtgagcgga taacaattcc ccggatccgg aggagggagg acaagctaca agacgtgcca
     3301 caacgtgcga gcgtgtacga acccgctctg tcgcatggtg tggaatagag acgtcaacgc
     3361 agctcgtaac atagcttgga tctgtatga
//
