LOCUS       sequence-418049-        4896 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express MmFnuc with pureexpress.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4896)
  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 4896)
  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: pKJ217
FEATURES             Location/Qualifiers
     source          1..4896
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        42..146
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(147..215,216..1007)
                     /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(365..384)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1178..1766
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1667..1686
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1920..1937
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2054..2075
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2090..2095,2096..2113,2114..2120)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2128..2144
                     /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     2133..2155
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2152..2168
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2152..2168
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        join(2189..2194,2195..2211,2212..2217)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     terminator      complement(2583..2630)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     2626..2644
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     RBS             4151..4173
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(4204..4223)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4205..4223)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4307..4326)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(4446..4463)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4446..4462)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4455..4477)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(4673..4692)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4792..4814
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4852..4870)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
       61 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
      121 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
      181 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
      241 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
      301 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
      361 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
      421 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
      481 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
      541 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
      601 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
      661 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
      721 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
      781 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
      841 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
      901 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
      961 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     1021 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     1081 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     1141 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     1201 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     1261 aagagctacc aactcttttt ccgaaggtaa ctggcttcag tagagcgcag ataccaaata
     1321 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     1381 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     1441 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     1501 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     1561 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     1621 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     1681 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     1741 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     1801 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     1861 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     1921 gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc
     1981 gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg
     2041 agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta
     2101 tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca
     2161 gctatgacat gattacgaat tctcatgttt gacagcttat catcgataag ctttaatgcg
     2221 gtagtttatc acagttaaat tgctaacgca gtcaggcacc gtgtatgaaa tctaacaatg
     2281 cgctcatcgt catcctcggc accgtcaccc tggatgctgt aggcataggc ttggttatgc
     2341 cggtactgcc gggcctcttg cgggatatcg tccattccga cagcatcgcc agtcactatg
     2401 gcgtgctgct agcgctatat gcgttgatgc aatttctatg cgcacccgtt ctcggagcac
     2461 tgtccgaccg ctttggccgc cgcccagtcc tgctcgcttc gctacttgga gccactatcg
     2521 actacgcgat catggcgacc acacccgtcc tgtggatatc cggatatagt tcctcctttc
     2581 agcaaaaaac ccctcaagac ccgtttagag gccccaaggg gttatgctag ttattgctca
     2641 gcggtggcag cagccaactc agcttccttt cgggctttgt tagcagccgg atctcctaag
     2701 gctcgagtta agcagcgaaa gggaaaaggc caagatactt acataaaata ttgcgggcac
     2761 catttacatc ccggtgggtt ttgtaattgc aggattcgca gataaaggtc tccttcccac
     2821 caagtttatt gtttttaacc ccacagtttc cacaggtctt ggatgtgtag cgttcgtctt
     2881 gagctattgc ctcacagtcg gttccgcgaa cccggtgaag cagtctttgc cggaaccgat
     2941 aatgctgcca acataacaag tctctattgg ctttcttggc aagatgattg ggtctgtcct
     3001 tgcgaacgat actgctgacg ctgatcttgc ccaggacgac cagcttataa tttttgacaa
     3061 ggtaactgca caacttaaga tggacgtcat caacgtaggt ttttaacttt aaatgaagtt
     3121 tcttaagctg tccaatacga tcttgcaccg cttgcgtttg ctttttcttt tgagcgtacg
     3181 ccaaaagacg gtgaacatag tctatcttct cgtgatattt atgaataact tctttcttgt
     3241 cggcttcagg cccaatctga aacgctttaa tgttgcttgg gtcatagcac gtagcaaacg
     3301 tccgtccccc tgggtcaatg ctgcagatac tatcagacgt atgcacctgg atttgtctgg
     3361 tataaatagg gtcgcaaggg atctgcagga cgaactttcc gttcggccgt ttgcatactt
     3421 tcatgtcgtg ggacagcgga ggtatctttg agacattttt acttaaccgc agaaaccgtt
     3481 ctttccagtc ttttttagat ttagaaaaat tatctgccat cagcgctatt ctactctgac
     3541 ggatccgctc gtcgggtatg tccttctctg tcaaaagtct aacatacttc ttttgcactt
     3601 caaacgatcc ctcacgtaac attgcgatat ctttatttct aaggtcgact ttggtcttct
     3661 tttggttggt ctgagcactt ttgtacatgg tacagaagtt cttgcatgcg gtcaacttta
     3721 ttgatgacat cttattgtca aaaaaccagt catctcctgg caatctatat tcagctggta
     3781 catcagtact attcgtttta gttacaacgc gctgaaggtc gaactgcttt ggtttgaaat
     3841 ctttctcttt aactaaataa ttacaccagt tataggcgtg gttagatacc tttaacatct
     3901 ggttaagggt tctcttttgc ttggatgttg gtctgatgcc aacactgaat gtacacatga
     3961 acttgtcctc cattttagca cattttgtct tcaggttaag atcagaataa tgagtactgg
     4021 atgcggagct aacgtcgcaa ttaagttcaa gcatacgagc cttatcaatc catgatttaa
     4081 atgtctcttg gccgtttaca aacgccgcct tccacaaagt catttgttct cttttccgct
     4141 tcatggtata tctccttctt aaagttaaac aaaattattt ctagagggaa accgttgtgg
     4201 tctccctata gtgagtcgta ttaatttcgc gggatcgaga tctcgatcct ctacgccgga
     4261 cgcatcgtgg ccggcatcac cggcgccaca ggtgcggttg ctggcgccta tatcgccgac
     4321 atcaccgatg gggaagatcg ggctcgccac ttcgggctca tgagcgcttg tttcggcgtg
     4381 ggtatggtgg caggccccgt ggccggggga ctgttgggcg ccatctcctt gcatgcaagc
     4441 ttggcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt
     4501 aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc
     4561 gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggcgcctgat gcggtatttt
     4621 ctccttacgc atctgtgcgg tatttcacac cgcatatatg gtgcactctc agtacaatct
     4681 gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct
     4741 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
     4801 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga
     4861 tacgcctatt tttataggtt aatgtcatga taataa
//
