LOCUS       sequence-429512-        6313 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammlian expression of DpFnuc for genome editing.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6313)
  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 6313)
  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: pKJ234
FEATURES             Location/Qualifiers
     source          1..6313
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        6..385
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        387..660
                     /label=chicken beta-actin promoter
     intron          661..1677
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     CDS             3157..3204
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     primer_bind     complement(3232..3249)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3238..3462
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3534..3551)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3534..3550)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3543..3565)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(3691..4146)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3828..3849)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4040..4059
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4173..4277
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4278..4346,4347..5138)
                     /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(4496..4515)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5309..5897
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5798..5817
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6051..6068
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6185..6206
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6221..6226,6227..6244,6245..6251)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6259..6275
                     /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)."
     primer_bind     6264..6286
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6283..6299
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6283..6299
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 gattcgacat tgattattga ctagttatta atagtaatca attacggggt cattagttca
       61 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc
      121 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
      181 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
      241 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc
      301 cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta
      361 cgtattagtc atcgctatta ccatggtcga ggtgagcccc acgttctgct tcactctccc
      421 catctccccc ccctccccac ccccaatttt gtatttattt attttttaat tattttgtgc
      481 agcgatgggg gcgggggggg ggggcgcgcg ccaggcgggg cggggcgggg cgaggggcgg
      541 ggcggggcga ggcggagagg tgcggcggca gccaatcaga gcggcgcgct ccgaaagttt
      601 ccttttatgg cgaggcggcg gcggcggcgg ccctataaaa agcgaagcgc gcggcgggcg
      661 ggagtcgctg cgcgctgcct tcgccccgtg ccccgctccg ccgccgcctc gcgccgcccg
      721 ccccggctct gactgaccgc gttactccca caggtgagcg ggcgggacgg cccttctcct
      781 ccgggctgta attagcgctt ggtttaatga cggcttgttt cttttctgtg gctgcgtgaa
      841 agccttgagg ggctccggga gggccctttg tgcgggggga gcggctcggg gggtgcgtgc
      901 gtgtgtgtgt gcgtggggag cgccgcgtgc ggctccgcgc tgcccggcgg ctgtgagcgc
      961 tgcgggcgcg gcgcggggct ttgtgcgctc cgcagtgtgc gcgaggggag cgcggccggg
     1021 ggcggtgccc cgcggtgcgg ggggggctgc gaggggaaca aaggctgcgt gcggggtgtg
     1081 tgcgtggggg ggtgagcagg gggtgtgggc gcgtcggtcg ggctgcaacc ccccctgcac
     1141 ccccctcccc gagttgctga gcacggcccg gcttcgggtg cggggctccg tacggggcgt
     1201 ggcgcggggc tcgccgtgcc gggcgggggg tggcggcagg tgggggtgcc gggcggggcg
     1261 gggccgcctc gggccgggga gggctcgggg gaggggcgcg gcggcccccg gagcgccggc
     1321 ggctgtcgag gcgcggcgag ccgcagccat tgccttttat ggtaatcgtg cgagagggcg
     1381 cagggacttc ctttgtccca aatctgtgcg gagccgaaat ctgggaggcg ccgccgcacc
     1441 ccctctagcg ggcgcggggc gaagcggtgc ggcgccggca ggaaggaaat gggcggggag
     1501 ggccttcgtg cgtcgccgcg ccgccgtccc cttctccctc tccagcctcg gggctgtccg
     1561 cggggggacg gctgccttcg ggggggacgg ggcagggcgg ggttcggctt ctggcgtgtg
     1621 actggcggct ctagagcctc tgctaaccat gttcatgcct tcttcttttt cctacagatc
     1681 cttaattaag ccaccatgaa gagaaagaga gaggatctga ccctgtggga tgcagctaac
     1741 gtgcataagc ataagagcat gtggtattgg tgggaataca ttcgccgcaa ggatatggtc
     1801 aatcacgaga aaacagactg tgatgtgatt cagttgcttc aatcagcctc agtcaagaaa
     1861 cagaaaaccc agagtgacaa gttcctcaca tctttctccg tggggatacg gccaacaaag
     1921 caccaaaaaa gggtgctcaa cgagatgttg agagtatcta attatacgta caattggtgc
     1981 ttgtggctcg taaacgagaa gggtctcaaa ccacaccagt ttgagttgca gaaaatcgtc
     2041 tgcaaaacaa atgctaatga tgttgacccg caatatcgaa tggagaatga cgactggttt
     2101 ttcaacaaca agatgacctc agttaagctg acgtcctgta agaacttttg cacttcttac
     2161 aaatcagcaa agagcctgaa gagcaagctt aagcgaccta tgtccgtatc aaacattatc
     2221 cagggctctt tttgtgtccc gaagcttttt attcgacatc tgtcctctaa ggatgtaagc
     2281 acggataaca caaatatgca gaacagatac atctgtatga tgcccgataa ctttgaaaaa
     2341 agatcaaacc cgaaggagag attcctgaaa ctcgcgaaac caatcactaa aatcccgccg
     2401 atcgatcacg atgtcaaaat cgtcaagcgc gccgatggca tgttcatcat gaacatacca
     2461 tgtgatccca agtatacaag aaggaatgca tcaaacgata ctattgagaa gcgggtttgt
     2521 ggaatagacc ctggggggcg aacttttgcc accgtttacg accccattga ctgctgtgtg
     2581 tttcaagtcg gaataaagga agataagcaa tatgtaataa gcaagctgca taataaaatc
     2641 gaccacgcgc atatgcacct tacaaaagcc cagaacaaaa aacagcaaca ggcggcacgg
     2701 gaaaggatcg tatctctcaa aaagacccat ttgaaattga aaactttcgt agatgatata
     2761 cacttgaaac tgtcttcaca tctggtaaaa gaatatcagt acgtcgccct cggtaagatc
     2821 aacgtagccc aattggtcaa aaccgatcga ccgaaaccgt tgtccaagag ggccaagcga
     2881 gacttgctgt attggcaaca ctataggttc cgacagaggc ttacgcacag gacaactaac
     2941 accgaatgca tacttgatgt gcaaaatgag gcgtacacat ctaaaacttg tggggtctgt
     3001 ggcaccatca ataagaattt ggaaaaaagt gaaacgttct actgtgatca gtgcaaatac
     3061 aacactcata gagatgttaa tggggccagg aatattcttt tgaaaagctt gcgcatgttc
     3121 cctttcgaaa aacagcaaca aggaggtggg ggctctaaaa ggccagctgc tacgaagaag
     3181 gccgggcagg ctaagaagaa aaaataacct aggtcagagc tcgctgatca gcctcgactg
     3241 tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg
     3301 aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga
     3361 gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg
     3421 aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa
     3481 ccagctgggg ctcgagatcc actagttcta gcctcgaggc tagagcggcc gccactggcc
     3541 gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca
     3601 gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc
     3661 caacagttgc gcagcctgaa tggcgaatgg aaattgtaag cgttaatatt ttgttaaaat
     3721 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa
     3781 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca
     3841 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
     3901 gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta
     3961 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg
     4021 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa
     4081 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
     4141 gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     4201 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     4261 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     4321 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     4381 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     4441 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     4501 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     4561 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     4621 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     4681 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     4741 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     4801 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     4861 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     4921 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     4981 ccggtgagcg tggctctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     5041 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     5101 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     5161 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     5221 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     5281 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     5341 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     5401 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     5461 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     5521 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     5581 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     5641 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     5701 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     5761 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     5821 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     5881 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     5941 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     6001 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     6061 tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga
     6121 ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg
     6181 caattaatgt gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg
     6241 ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc
     6301 atgaggcgcg ccg
//
