LOCUS       sequence-442959-        2483 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammlian expression of DpFnuc guide RNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2483)
  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 2483)
  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: pKJ248
FEATURES             Location/Qualifiers
     source          1..2483
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     290..309
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     ncRNA           complement(493..560)
                     /product="hepatitis delta virus self-cleaving ribozyme 
                     (Ferre-D'Amare and Scott, 2010)"
                     /label=HDV ribozyme
                     /ncRNA_class="ribozyme"
     promoter        complement(793..1041)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(851..870)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(1021..1041)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     promoter        1148..1252
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1253..1321,1322..2113)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1471..1490)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(2284..2483,1..389)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
       61 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
      121 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
      181 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
      241 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
      301 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
      361 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
      421 tggccttttg ctggcctttt gctcacatgt catatgtaga gaggtacctc gagcggccca
      481 agcttaaaaa aagtcccatt cgccatgccg aagcatgttg cccagccggc gccagcgagg
      541 aggctgggac catgccggcc taaccattat tgacatataa taacccattt cattcggcag
      601 aacattgccc tggactaagc atacgcaaag tctcgtagca atggactttc gaatttagga
      661 tggatcgacc taattgttac agagtaatta taactgcgaa ttgacaaatg gaaacatgcg
      721 caacgatttt agcaatatat ttctagcacc attaacgtct ctgtgtgtgt tatatttgca
      781 gttatcgcaa tcggtgtttc gtcctttcca caagatatat aaagccaaga aatcgaaata
      841 ctttcaagtt acggtaagca tatgatagtc cattttaaaa cataatttta aaactgcaaa
      901 ctacccaaga aattattact ttctacgtca cgtattttgt actaatatct ttgtgtttac
      961 agtcaaatta attctaatta tctctctaac agccttgtat cgtatatgca aatatgaagg
     1021 aatcatggga aataggccct cttcctgccc gaccttggta ccggatccag tcgactgaat
     1081 tggttccttt aaagcctgct tttttgtaca gctagcgacg tcaggtggca cttttcgggg
     1141 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     1201 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     1261 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     1321 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     1381 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     1441 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     1501 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     1561 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     1621 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     1681 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     1741 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     1801 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     1861 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     1921 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtggat ctcgcggtat
     1981 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     2041 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     2101 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     2161 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     2221 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     2281 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     2341 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     2401 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     2461 cttcaagaac tctgtagcac cgc
//
