LOCUS       sequence-234700-        5978 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses concatenated noncoding sequences surrounding the 
            CRISPR-Cas12c2 system in the native genomic loci; expresses the 
            tracrRNA required for activity..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5978)
  AUTHORS   Yan WX, Hunnewell P, Alfonse LE, Carte JM, Keston-Smith E, 
            Sothiselvam S, Garrity AJ, Chong S, Makarova KS, Koonin EV, Cheng 
            DR, Scott DA
  TITLE     Functionally diverse type V CRISPR-Cas systems.
  JOURNAL   Science. 2019 Jan 4;363(6422):88-91. doi: 10.1126/science.aav7271. 
            Epub 2018 Dec 6.
  PUBMED    30523077
REFERENCE   2  (bases 1 to 5978)
  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: 
            "10.1126/science.aav7271"; journalName: "Science"; date: "2019-01-4-
            4"; volume: "363"; issue: "6422"; pages: "88-91"
COMMENT     Sequence Label: pACYC-Cas12c2-noncoding
FEATURES             Location/Qualifiers
     source          1..5978
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(215..232)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(349..894)
                     /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(429..448)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        join(1006..1011,1012..1028,1029..1034)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     CDS             1082..2272
                     /codon_start=1
                     /gene="tet"
                     /product="tetracycline efflux protein"
                     /label=TcR
                     /note="confers resistance to tetracycline"
                     /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
                     GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
                     VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
                     LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
                     QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
                     AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
                     AALTSLTSIIGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
     primer_bind     complement(1434..1453)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     2711..2730
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2711..2729
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2730..2754
                     /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)."
     promoter        join(2763..2768,2769..2786,2787..2792)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(4357..4375)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4371..4418
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(5039..5698)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     5613..5632
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(5699..5801)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
ORIGIN
        1 ggcttctgtt tctatcagct gtccctcctg ttcagctact gacggggtgg tgcgtaacgg
       61 caaaagcacc gccggacatc agcgctagcg gagtgtatac tggcttacta tgttggcact
      121 gatgagggtg tcagtgaagt gcttcatgtg gcaggagaaa aaaggctgca ccggtgcgtc
      181 agcagaatat gtgatacagg atatattccg cttcctcgct cactgactcg ctacgctcgg
      241 tcgttcgact gcggcgagcg gaaatggctt acgaacgggg cggagatttc ctggaagatg
      301 ccaggaagat acttaacagg gaagtgagag ggccgcggca aagccgtttt tccataggct
      361 ccgcccccct gacaagcatc acgaaatctg acgctcaaat cagtggtggc gaaacccgac
      421 aggactataa agataccagg cgtttccccc tggcggctcc ctcgtgcgct ctcctgttcc
      481 tgcctttcgg tttaccggtg tcattccgct gttatggccg cgtttgtctc attccacgcc
      541 tgacactcag ttccgggtag gcagttcgct ccaagctgga ctgtatgcac gaaccccccg
      601 ttcagtccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggaaagac
      661 atgcaaaagc accactggca gcagccactg gtaattgatt tagaggagtt agtcttgaag
      721 tcatgcgccg gttaaggcta aactgaaagg acaagttttg gtgactgcgc tcctccaagc
      781 cagttacctc ggttcaaaga gttggtagct cagagaacct tcgaaaaacc gccctgcaag
      841 gcggtttttt cgttttcaga gcaagagatt acgcgcagac caaaacgatc tcaagaagat
      901 catcttatta atcagataaa atatttctag atttcagtgc aatttatctc ttcaaatgta
      961 gcacctgaag tcagccccat acgatataag ttgtaattct catgtttgac agcttatcat
     1021 cgataagctt taatgcggta gtttatcaca gttaaattgc taacgcagtc aggcaccgtg
     1081 tatgaaatct aacaatgcgc tcatcgtcat cctcggcacc gtcaccctgg atgctgtagg
     1141 cataggcttg gttatgccgg tactgccggg cctcttgcgg gatatcgtcc attccgacag
     1201 catcgccagt cactatggcg tgctgctagc gctatatgcg ttgatgcaat ttctatgcgc
     1261 acccgttctc ggagcactgt ccgaccgctt tggccgccgc ccagtcctgc tcgcttcgct
     1321 acttggagcc actatcgact acgcgatcat ggcgaccaca cccgtcctgt ggatcctcta
     1381 cgccggacgc atcgtggccg gcatcaccgg cgccacaggt gcggttgctg gcgcctatat
     1441 cgccgacatc accgatgggg aagatcgggc tcgccacttc gggctcatga gcgcttgttt
     1501 cggcgtgggt atggtggcag gccccgtggc cgggggactg ttgggcgcca tctccttgca
     1561 tgcaccattc cttgcggcgg cggtgctcaa cggcctcaac ctactactgg gctgcttcct
     1621 aatgcaggag tcgcataagg gagagcgtcg accgatgccc ttgagagcct tcaacccagt
     1681 cagctccttc cggtgggcgc ggggcatgac tatcgtcgcc gcacttatga ctgtcttctt
     1741 tatcatgcaa ctcgtaggac aggtgccggc agcgctctgg gtcattttcg gcgaggaccg
     1801 ctttcgctgg agcgcgacga tgatcggcct gtcgcttgcg gtattcggaa tcttgcacgc
     1861 cctcgctcaa gccttcgtca ctggtcccgc caccaaacgt ttcggcgaga agcaggccat
     1921 tatcgccggc atggcggccg acgcgctggg ctacgtcttg ctggcgttcg cgacgcgagg
     1981 ctggatggcc ttccccatta tgattcttct cgcttccggc ggcatcggga tgcccgcgtt
     2041 gcaggccatg ctgtccaggc aggtagatga cgaccatcag ggacagcttc aaggatcgct
     2101 cgcggctctt accagcctaa cttcgatcat tggaccgctg atcgtcacgg cgatttatgc
     2161 cgcctcggcg agcacatgga acgggttggc atggattgta ggcgccgccc tataccttgt
     2221 ctgcctcccc gcgttgcgtc gcggtgcatg gagccgggcc acctcgacct gaatggaagc
     2281 cggcggcacc tcgctaacgg attcaccact ccaagaattg gagccaatca attcttgcgg
     2341 agaactgtga atgcgcaaac caacccttgg cagaacatat ccatcgcgtc cgccatctcc
     2401 agcagccgca cgcggcgcat ctcgggcagc gttgggtcct ggccacgggt gcgcatgatc
     2461 gtgctcctgt cgttgaggac ccggctaggc tggcggggtt gccttactgg ttagcagaat
     2521 gaatcaccga tacgcgagcg aacgtgaagc gactgctgct gcaaaacgtc tgcgacctga
     2581 gcaacaacat gaatggtctt cggtttccgt gtttcgtaaa gtctggaaac gcggaagtcc
     2641 cctacgtgct gctgaagttg cccgcaacag agagtggaac caaccggtga taccacgata
     2701 ctatgactga taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta
     2761 gatttacact ttatgcttcc ggctcgtatg tttctagact cgagatgctg gccaaggaat
     2821 ttggtttgtc ggatgtaatg attgggaagc gatgcaaaaa actgggtgtt gtaaaaccaa
     2881 agccgggatt ctgggcgaag gtggacgctg ggaaagtgcc acatccccaa ggaaagaaag
     2941 tagccgggca gtaaataacg ctttggaata tgagacgtat ctgtagaatt tttggtatcc
     3001 agtaaaaaga gcaaaacgaa atgacaaaac actcaatacc ccttcacgca ttccgcaact
     3061 ccggcgcaga cgccagaaaa tggaaaggcc gcattgcttt gctggccaag cgcggcaaag
     3121 aaacgatgcg aacactgcag tttccgctcg aaatgagcga gcctgaagca ctgcgccgtg
     3181 cgcccaagaa tcgccgcacc aaggacacca ccgtaagcga gtatcactgt gtcttttcag
     3241 actgcggaaa gacaatgcac gcggatgaga atgcggcagt gaatatcggc ggaaagttca
     3301 ttgcagatat cgaaaaatag tttgcacaaa gtgcgagaaa agatataata cacacatacc
     3361 acccgtgcat ttctggatca atgatccgta cctcaatgtc cgggcgcgca gctagagcga
     3421 cctgaaatct gcacgaaaac cggcgaaagc cggttttttg ttgcctgtag tttgagttta
     3481 tttctttgcg cctgggatgg caaagccagc tttcaccggg cgaccgccga tacgctcacc
     3541 ttcgactggg atcacaagcc ttgccgtttc cttggcgaga tagctgcgca gtgccaacac
     3601 ttccgcatgc aagacatttt tacgcctacc acctttgtac acgaacttat agcgagcttt
     3661 ggatttttgg ataggttcaa tcgaatttag gtagggtatc caaagtgccg ctcgaccacc
     3721 acggatgttt ttccatgtcc aaatagagtc tggatgttga aattgtgaga tttccatatg
     3781 caagctgggt ggctgttaaa aatgaatttc atgataatac ataatgttat attatgaacg
     3841 acttcccccg cgtcactagc aggattcagg ttgggtttga ggggtcagcg tactcaggaa
     3901 gcaggattca ggttgggttt gaggagctgg atgccgttcc acagcaggat tcaggttggg
     3961 tttgaggggc tggccagaac aagaaagcag gattcaggtt gggtttgagg ctgtctcctt
     4021 ttggaaaagc aggattcagg ttgggtttga gggctcaatc ctattcagaa agcaggattc
     4081 aggttgggtt tgaggtggcc aggagaacag aaagcaggat tcaggttggg tttgaggtgt
     4141 gcatgaggat gctaaagcag gattcaggtt gggtttgagg ctagcgggat tacaacaaag
     4201 caggatttag gttggatttg agggtgccat caaaatttgg tagcaggatt caggttgggg
     4261 ttgaggccga aaagattcag atgagcagga ttcaggttgg gtttgaggct cgagctgcta
     4321 acaaagcccg aaaggaagct gagttggctg ctgccaccgc tgagcaataa ctagcataac
     4381 cccttggggc ctctaaacgg gtcttgaggg gttttttgct gaaaggagga actatatccg
     4441 gatgagtcaa cgccatgagc ggcctcattt cttattctga gttacaacag tccgcaccgc
     4501 tgccggtagc tccttccggt gggcgcgggg catgactatc gtcgccgcac ttatgactgt
     4561 cttctttatc atgcaactcg taggacaggt gccggcagcg cccaacagtc ccccggccac
     4621 ggggcctgcc accataccca cgccgaaaca agcgccctgc accattatgt tccggatctg
     4681 catcgcagga tgctgctggc taccctgtgg aacacctaca tctgtattaa cgaagcgcta
     4741 accgttttta tcaggctctg ggaggcagaa taaatgatca tatcgtcaat tattacctcc
     4801 acggggagag cctgagcaaa ctggcctcag gcatttgaga agcacacggt cacactgctt
     4861 ccggtagtca ataaaccggt aaaccagcaa tagacataag cggctattta acgaccctgc
     4921 cctgaaccga cgaccgggtc gaatttgctt tcgaatttct gccattcatc cgcttattat
     4981 cacttattca ggcgtagcaa ccaggcgttt aagggcacca ataactgcct taaaaaaatt
     5041 acgccccgcc ctgccactca tcgcagtact gttgtaattc attaagcatt ctgccgacat
     5101 ggaagccatc acaaacggca tgatgaacct gaatcgccag cggcatcagc accttgtcgc
     5161 cttgcgtata atatttgccc atggtgaaaa cgggggcgaa gaagttgtcc atattggcca
     5221 cgtttaaatc aaaactggtg aaactcaccc agggattggc tgagacgaaa aacatattct
     5281 caataaaccc tttagggaaa taggccaggt tttcaccgta acacgccaca tcttgcgaat
     5341 atatgtgtag aaactgccgg aaatcgtcgt ggtattcact ccagagcgat gaaaacgttt
     5401 cagtttgctc atggaaaacg gtgtaacaag ggtgaacact atcccatatc accagctcac
     5461 cgtctttcat tgccatacgg aattccggat gagcattcat caggcgggca agaatgtgaa
     5521 taaaggccgg ataaaacttg tgcttatttt tctttacggt ctttaaaaag gccgtaatat
     5581 ccagctgaac ggtctggtta taggtacatt gagcaactga ctgaaatgcc tcaaaatgtt
     5641 ctttacgatg ccattgggat atatcaacgg tggtatatcc agtgattttt ttctccattt
     5701 tagcttcctt agctcctgaa aatctcgata actcaaaaaa tacgcccggt agtgatctta
     5761 tttcattatg gtgaaagttg gaacctctta cgtgccgatc aacgtctcat tttcgccaaa
     5821 agttggccca gggcttcccg gtatcaacag ggacaccagg atttatttat tctgcgaagt
     5881 gatcttccgt cacaggtatt tattcggcgc aaagtgcgtc gggtgatgct gccaacttac
     5941 tgatttagtg tatgatggtg tttttgaggt gctccagt
//
