LOCUS       sequence-234750-        5494 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses concatenated noncoding sequences surrounding the 
            CRISPR-OspCas12c 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 5494)
  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 5494)
  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-OspCas12c-noncoding
FEATURES             Location/Qualifiers
     source          1..5494
                     /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(3873..3891)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3887..3934
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(4555..5214)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     5129..5148
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(5215..5317)
                     /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 cgagatgcgc atacaaaacc
     2821 cattgcaatg gtagttatat agtgatacaa gtattggata tgaatgacga aaataaaaca
     2881 aaaagccacc caaattcaat ttggtattgg tgttctggtg agaaactatg gtatttgcct
     2941 tactctggga cacctaaaag aacatctaaa tctgtttgga cggtttctgg ttcttttggc
     3001 caactagata cattaatcca tgggcaggta attgctctgc gacattattt aaatggaaat
     3061 atgcagcgtt atttaccccc cgttgaagaa ctagcaaaag agcgggggcg taagcgagca
     3121 gtcagctata cactacctgg ccaaatttgg aaaaaataat catgacaaag ttaagacacc
     3181 gacagaaaaa attaacacac gactgggctg gctccaaaaa gagggaagta ttaggctcaa
     3241 atggcaagtt tcagaatccg ttgttaatgc cggttaaaaa aggtcaggtt actgagttcc
     3301 ggaaagcgtt tcattcacct gagtttatcg ctaaaaaacg cggcataggg tctgagtatt
     3361 tttgcgctta ttccgattgc aactcatcct taaattctga agctaacgca gctgctaacg
     3421 tagcgcaaaa atttcaaaaa cagttatttt ttgagttata aaatgcttaa taaaaatcgc
     3481 ttaaaattac ttgttttctg gttctaatta ggtagtataa aggttcatcg ttggtatggc
     3541 tgtgttcttc tgaacgcaca tcattctctt agagacgtct acataccaac ccacgatggg
     3601 gagcgtggac aacaaaatct ccccactact ttctttttta gacttaacgt ttgttgtttg
     3661 ttatttttct actcttaatt tatatcttct atttttgttg cctgtgcttt cattcccccc
     3721 gcttccatag ttaaacctct gtttattatc gtacttattg ctacagcagc ctgtaatgcg
     3781 ttttccatgt gttgttttgc attcactggt tctgcatttc cgcactcgag ctgctaacaa
     3841 agcccgaaag gaagctgagt tggctgctgc caccgctgag caataactag cataacccct
     3901 tggggcctct aaacgggtct tgaggggttt tttgctgaaa ggaggaacta tatccggatg
     3961 agtcaacgcc atgagcggcc tcatttctta ttctgagtta caacagtccg caccgctgcc
     4021 ggtagctcct tccggtgggc gcggggcatg actatcgtcg ccgcacttat gactgtcttc
     4081 tttatcatgc aactcgtagg acaggtgccg gcagcgccca acagtccccc ggccacgggg
     4141 cctgccacca tacccacgcc gaaacaagcg ccctgcacca ttatgttccg gatctgcatc
     4201 gcaggatgct gctggctacc ctgtggaaca cctacatctg tattaacgaa gcgctaaccg
     4261 tttttatcag gctctgggag gcagaataaa tgatcatatc gtcaattatt acctccacgg
     4321 ggagagcctg agcaaactgg cctcaggcat ttgagaagca cacggtcaca ctgcttccgg
     4381 tagtcaataa accggtaaac cagcaataga cataagcggc tatttaacga ccctgccctg
     4441 aaccgacgac cgggtcgaat ttgctttcga atttctgcca ttcatccgct tattatcact
     4501 tattcaggcg tagcaaccag gcgtttaagg gcaccaataa ctgccttaaa aaaattacgc
     4561 cccgccctgc cactcatcgc agtactgttg taattcatta agcattctgc cgacatggaa
     4621 gccatcacaa acggcatgat gaacctgaat cgccagcggc atcagcacct tgtcgccttg
     4681 cgtataatat ttgcccatgg tgaaaacggg ggcgaagaag ttgtccatat tggccacgtt
     4741 taaatcaaaa ctggtgaaac tcacccaggg attggctgag acgaaaaaca tattctcaat
     4801 aaacccttta gggaaatagg ccaggttttc accgtaacac gccacatctt gcgaatatat
     4861 gtgtagaaac tgccggaaat cgtcgtggta ttcactccag agcgatgaaa acgtttcagt
     4921 ttgctcatgg aaaacggtgt aacaagggtg aacactatcc catatcacca gctcaccgtc
     4981 tttcattgcc atacggaatt ccggatgagc attcatcagg cgggcaagaa tgtgaataaa
     5041 ggccggataa aacttgtgct tatttttctt tacggtcttt aaaaaggccg taatatccag
     5101 ctgaacggtc tggttatagg tacattgagc aactgactga aatgcctcaa aatgttcttt
     5161 acgatgccat tgggatatat caacggtggt atatccagtg atttttttct ccattttagc
     5221 ttccttagct cctgaaaatc tcgataactc aaaaaatacg cccggtagtg atcttatttc
     5281 attatggtga aagttggaac ctcttacgtg ccgatcaacg tctcattttc gccaaaagtt
     5341 ggcccagggc ttcccggtat caacagggac accaggattt atttattctg cgaagtgatc
     5401 ttccgtcaca ggtatttatt cggcgcaaag tgcgtcgggt gatgctgcca acttactgat
     5461 ttagtgtatg atggtgtttt tgaggtgctc cagt
//
