LOCUS       sequence-383604-        5858 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes MmCas12m under a constitutive promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5858)
  AUTHORS   Wu WY, Mohanraju P, Liao C, Adiego-Perez B, Creutzburg SCA, Makarova
            KS, Keessen K, Lindeboom TA, Khan TS, Prinsen S, Joosten R, Yan WX, 
            Migur A, Laffeber C, Scott DA, Lebbink JHG, Koonin EV, Beisel CL, 
            van der Oost J
  TITLE     The miniature CRISPR-Cas12m effector binds DNA to block 
            transcription.
  JOURNAL   Mol Cell. 2022 Dec 1;82(23):4487-4502.e7. doi: 
            10.1016/j.molcel.2022.11.003. Epub 2022 Nov 24.
  PUBMED    36427491
REFERENCE   2  (bases 1 to 5858)
  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"; doi: 
            "10.1016/j.molcel.2022.11.003"; journalName: "Mol Cell"; date: 
            "2022-12-1- 1"; volume: "82"; issue: "23"; pages: "4487-4502.e7"
COMMENT     Sequence Label: pCas-Cas12m
FEATURES             Location/Qualifiers
     source          1..5858
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(391..1050)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     965..984
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(1051..1153)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(1545..1562)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1679..2224)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(1759..1778)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2300..2319
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        2414..2448
                     /label=BBa_J23108
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.51"
     primer_bind     complement(4295..4312)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      4465..4551
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4643..4670
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        4689..4780
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5238..5693
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5325..5344)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5535..5556
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
       61 ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac
      121 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtc
      181 aggcatttga gaagcacacg gtcacactgc ttccggtagt caataaaccg gtaaaccagc
      241 aatagacata agcggctatt taacgaccct gccctgaacc gacgaccggg tcgaatttgc
      301 tttcgaattt ctgccattca tccgcttatt atcacttatt caggcgtagc aaccaggcgt
      361 ttaagggcac caataactgc cttaaaaaaa ttacgccccg ccctgccact catcgcagta
      421 ctgttgtaat tcattaagca ttctgccgac atggaagcca tcacaaacgg catgatgaac
      481 ctgaatcgcc agcggcatca gcaccttgtc gccttgcgta taatatttgc ccatggtgaa
      541 aacgggggcg aagaagttgt ccatattggc cacgtttaaa tcaaaactgg tgaaactcac
      601 ccagggattg gctgagacga aaaacatatt ctcaataaac cctttaggga aataggccag
      661 gttttcaccg taacacgcca catcttgcga atatatgtgt agaaactgcc ggaaatcgtc
      721 gtggtattca ctccagagcg atgaaaacgt ttcagtttgc tcatggaaaa cggtgtaaca
      781 agggtgaaca ctatcccata tcaccagctc accgtctttc attgccatac ggaattccgg
      841 atgagcattc atcaggcggg caagaatgtg aataaaggcc ggataaaact tgtgcttatt
      901 tttctttacg gtctttaaaa aggccgtaat atccagctga acggtctggt tataggtaca
      961 ttgagcaact gactgaaatg cctcaaaatg ttctttacga tgccattggg atatatcaac
     1021 ggtggtatat ccagtgattt ttttctccat tttagcttcc ttagctcctg aaaatctcga
     1081 taactcaaaa aatacgcccg gtagtgatct tatttcatta tggtgaaagt tggaacctct
     1141 tacgtgccga tcaacgtctc attttcgcca aaagttggcc cagggcttcc cggtatcaac
     1201 agggacacca ggatttattt attctgcgaa gtgatcttcc gtcacaggta tttattcggc
     1261 gcaaagtgcg tcgggtgatg ctgccaactt actgatttag tgtatgatgg tgtttttgag
     1321 gtgctccagt ggcttctgtt tctatcagct gtccctcctg ttcagctact gacggggtgg
     1381 tgcgtaacgg caaaagcacc gccggacatc agcgctagcg gagtgtatac tggcttacta
     1441 tgttggcact gatgagggtg tcagtgaagt gcttcatgtg gcaggagaaa aaaggctgca
     1501 ccggtgcgtc agcagaatat gtgatacagg atatattccg cttcctcgct cactgactcg
     1561 ctacgctcgg tcgttcgact gcggcgagcg gaaatggctt acgaacgggg cggagatttc
     1621 ctggaagatg ccaggaagat acttaacagg gaagtgagag ggccgcggca aagccgtttt
     1681 tccataggct ccgcccccct gacaagcatc acgaaatctg acgctcaaat cagtggtggc
     1741 gaaacccgac aggactataa agataccagg cgtttccccc tggcggctcc ctcgtgcgct
     1801 ctcctgttcc tgcctttcgg tttaccggtg tcattccgct gttatggccg cgtttgtctc
     1861 attccacgcc tgacactcag ttccgggtag gcagttcgct ccaagctgga ctgtatgcac
     1921 gaaccccccg ttcagtccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     1981 ccggaaagac atgcaaaagc accactggca gcagccactg gtaattgatt tagaggagtt
     2041 agtcttgaag tcatgcgccg gttaaggcta aactgaaagg acaagttttg gtgactgcgc
     2101 tcctccaagc cagttacctc ggttcaaaga gttggtagct cagagaacct tcgaaaaacc
     2161 gccctgcaag gcggtttttt cgttttcaga gcaagagatt acgcgcagac caaaacgatc
     2221 tcaagaagat catcttatta atcagataaa atatttctag gctcatgagc ccgaagtggc
     2281 gagcccgatc ttccccatcg gtgatgtcgg cgatataggc gccagcaacc gcacctgtgg
     2341 cgctggtgat gccggccacg atgcgtccgg cgtagaggat ctgctcatgt ttgacagctt
     2401 atcatcgatg catctgacag ctagctcagt cctaggtata atgctagcga gctcaggagg
     2461 acatcatgac aacaatgaca gtacatacaa tgggagtaca ttataagtgg cagatccccg
     2521 aagtccttcg gcagcaactg tggttagcac ataacctgcg ggaagatctg gtatcgcttc
     2581 agttagcata tgatgatgat ttaaaggcaa tttggtcatc gtatcccgac gtagcacagg
     2641 cagaagatac aatggcagca gccgaagcag atgcagttgc actgtcggaa cgggttaaac
     2701 aggcacggat tgaagcacgg tcgaaaaaga tttcgacaga actgacacaa cagttacggg
     2761 atgcaaaaaa acggttaaaa gatgcccggc aagcacggcg ggatgcaatt gcagttgtaa
     2821 aggatgacgc agccgaacgg cggaaggcac ggtcggatca attagcagca gatcagaagg
     2881 cactgtatgg gcaatattgc cgagatggag atctgtattg ggcatcgttt aatacagtac
     2941 tggatcacca taaaacagcc gttaagcgaa tcgcagcaca acgggcctca gggaaaccgg
     3001 caactcttcg acaccaccgg tttgacggat cgggaacaat tgcagtacag ctgcagcggc
     3061 aggcaggggc accgccgcgg acacccatgg tcttagcaga tgaagcagga aaatatcgga
     3121 atgtacttca tatccccggg tggactgatc ccgacgtatg ggaacagatg acacggtcac
     3181 aatgccggca atcggggcgg gttacagtac ggatgcggtg cggatcgaca gatggacagc
     3241 cccagtggat tgatctaccg gtacaagtac atcgctggtt accggcagat gcagatatta
     3301 caggagcaga attagttgtc acacgggttg caggaattta tcgggcaaaa ctgtgtgtta
     3361 cagcacggat tggggatact gaacccgtta catcggggcc gacagtagca ttacatctgg
     3421 gatggcgctc gacagaagaa ggaacagcag tagcaacttg gcggtcagat gcccccctgg
     3481 atattccctt tggactacgg acagtaatgc gggttgacgc cgcagggaca tcggggatta
     3541 ttgtagtacc cgcaacaatt gaacggcggc tgactcggac tgaaaatatt gcatcgtcgc
     3601 ggtcgctggc actagacgca ctacgggata aggttgtcgg gtggctatca gataacgatg
     3661 cccccacata ccgggatgcc ccgctggaag cagccactgt taagcagtgg aagtcacccc
     3721 agcgctttgc ctcgctagca catgcatgga aggataatgg aacagaaatt tcggatattt
     3781 tatgggcatg gttttcgtta gatcgcaaac aatgggcaca acaagaaaat gggcggcgga
     3841 aagccttagg gcatcgggat gatttatatc ggcagattgc agcagtaatt tcggatcagg
     3901 caggacatgt tttagttgat gatacatcgg tagcagaatt atcggcacgg gcaatggaac
     3961 ggactgaatt acccacagaa gtacaacaga aaattgatcg gcggcgggac catgcagcac
     4021 ccggggggct acgggcatcg gttgtagcag caatgacacg cgatggagtc cccgtcacaa
     4081 ttgttgccgc cgcagacttt actcggactc attcgcgctg cggacatgtt aatcccgccg
     4141 atgatcggta tctgtcgaat cccgtacggt gcgacggatg cggggcaatg tatgatcaag
     4201 atcggtcgtt cgttactctg atgctgcgcg cagcaactgc cccctcgaac ccctagaagc
     4261 ttggctgttt tggcggatga gagaagattt tcagcctgat acagattaaa tcagaacgca
     4321 gaagcggtct gataaaacag aatttgcctg gcggcagtag cgcggtggtc ccacctgacc
     4381 ccatgccgaa ctcagaagtg aaacgccgta gcgccgatgg tagtgtgggg tctccccatg
     4441 cgagagtagg gaactgccag gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc
     4501 tttcgtttta tctgttgttt gtcggtgaac gctctcctga gtaggacaaa tccgccggga
     4561 gcggatttga acgttgcgaa gcaacggccc ggagggtggc gggcaggacg cccgccataa
     4621 actgccaggc atcaaattaa gcagaaggcc atcctgacgg atggcctttt tgcgtttcta
     4681 caaactcttt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     4741 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     4801 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     4861 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca gcaaactatt aactggcgaa
     4921 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     4981 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     5041 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     5101 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     5161 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     5221 atactttaga ttgatttacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     5281 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     5341 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     5401 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg atttgggtga
     5461 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     5521 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggg
     5581 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
     5641 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgtttacaa tttaaaagga
     5701 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     5761 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     5821 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccg
//
