LOCUS       sequence-428763-        3389 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express MmFnuc 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: pKJ221
FEATURES             Location/Qualifiers
     source          1..3389
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     410..427
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(451..508)
                     /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      999..1587
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1488..1507
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2036..2055
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2036..2054
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2054..2078
                     /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)."
     ncRNA           2408..2475
                     /product="hepatitis delta virus self-cleaving ribozyme 
                     (Ferre-D'Amare and Scott, 2010)"
                     /label=HDV ribozyme
                     /ncRNA_class="ribozyme"
     terminator      2484..2531
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(2856..2875)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             join(2867..3389,1..293)
                     /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             join(2867..3389,1..293)
                     /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(2940..2959)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
ORIGIN
        1 ttgaacaagt ctggaaagaa atgcataagc ttttgccatt ctcaccggat tcagtcgtca
       61 ctcatggtga tttctcactt gataacctta tttttgacga ggggaaatta ataggttgta
      121 ttgatgttgg acgagtcgga atcgcagacc gataccagga tcttgccatc ctatggaact
      181 gcctcggtga gttttctcct tcattacaga aacggctttt tcaaaaatat ggtattgata
      241 atcctgatat gaataaattg cagtttcatt tgatgctcga tgagtttttc taatcagaat
      301 tggttaattg gttgtaacac tggcagagca ttacgctgac ttgacgggac tcgagattac
      361 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
      421 gagcgaggaa gcctgcataa cgcgaagtaa tcttttcggt tttaaagaaa aagggcaggg
      481 tggtgacacc ttgccctttt ttgccggact gcagcaatgg caacaacgtt gcgcaaacta
      541 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
      601 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
      661 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
      721 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
      781 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
      841 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
      901 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
      961 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     1021 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     1081 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     1141 actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     1201 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     1261 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     1321 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     1381 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     1441 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     1501 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     1561 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     1621 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     1681 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     1741 agcgaggtcg gcgatatagg cgcgagcaac cgcccctgtg gcgccggtga tgccggccac
     1801 gatgcgtccg gcgtagagga tctgctcatg tttgacagct tatcatcgat gcataatgtg
     1861 cctgtcaaat ggacgaagca gggattctgc aaaccctatg ctactccctc gagccgtcaa
     1921 ttgtctgatt cgttaccaat tatactagct ccggcaaaaa aacgggcaag gtgtcaccac
     1981 cctgcccttt ttctttaaaa ccgaaaagat tacttcgcgt ttgccacctg acgtctaata
     2041 cgactcacta tagggaattg tgagcggata acaattcccc ggatccactt ccaagacctg
     2101 tggtaattgc ggtgtgaaga acaacaaact tggtggaaag gaaacgttta cttgtgagtg
     2161 ttgcaattac aaaactcatc gagacgtcaa cggagcgaga aacattctgt gcaaatactt
     2221 gaaacttttt ccattcgcag cataacgaaa gaaactgaca atcgattttt tcgggttcga
     2281 ttctatccca cttgactcaa agagtcagag ggctcgaata cattttctgc acaggttttg
     2341 ctaatgcaag atctggggca agaatgtgtt cgggtcaaat gagttagcag ccacctcctt
     2401 gttattgggc cggcatggtc ccagcctcct cgctggcgcc ggctgggcaa catgcttcgg
     2461 catggcgaat gggacgcggc cgcctagcat aaccccttgg ggcctctaaa cgggtcttga
     2521 ggggtttttt ggctagcgct gaggtctgcc tcgtgaagaa ggtgttgctg actcatacca
     2581 ggcctgaatc gccccatcat ccagccagaa agtgagggag ccacggttga tgagagcttt
     2641 gttgtaggtg gaccagttgg tgattttgaa cttttgcttt gccacggaac ggtctgcgtt
     2701 gtcgggaaga tgcgtgatct gatccttcaa ctcagcaaaa gttcgattta ttcaacaaag
     2761 ccacgttgtg tctcaaaatc tctgatgtta cattgcacaa gataaaaata tatcatcatg
     2821 aacaataaaa ctgtctgctt acataaacag taatacaagg ggtgttatga gccatattca
     2881 acgggaaacg tcttgctcca ggccgcgatt aaattccaac atggatgctg atttatatgg
     2941 gtataaatgg gctcgcgata atgtcgggca atcaggtgcg acaatctatc gattgtatgg
     3001 gaagcccgat gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg ccaatgatgt
     3061 tacagatgag atggtcagac taaactggct gacggaattt atgcctcttc cgaccatcaa
     3121 gcattttatc cgtactcctg atgatgcatg gttactcacc actgcgatcc ccgggaaaac
     3181 agcattccag gtattagaag aatatcctga ttcaggtgaa aatattgttg atgcgctggc
     3241 agtgttcctg cgccggttgc attcgattcc tgtttgtaat tgtcctttta acagcgatcg
     3301 cgtatttcgt ctcgctcagg cgcaatcacg aatgaataac ggtttggttg atgcgagtga
     3361 ttttgatgac gagcgtaatg gctggcctg
//
