LOCUS       sequence-373981-        7798 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of single-spacer CRISPR array with a shortened 
            non-targeting spacer for PbuCas13b and expression of PbuCas13b in 
            bacteria..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7798)
  AUTHORS   Wandera KG, Alkhnbashi OS, Bassett HVI, Mitrofanov A, Hauns S, Migur
            A, Backofen R, Beisel CL
  TITLE     Anti-CRISPR prediction using deep learning reveals an inhibitor of 
            Cas13b nucleases.
  JOURNAL   Mol Cell. 2022 May 24. pii: S1097-2765(22)00437-3. doi: 
            10.1016/j.molcel.2022.05.003.
  PUBMED    35649413
REFERENCE   2  (bases 1 to 7798)
  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: "Mol Cell. 
            2022 May 24. pii: S1097-2765(22)00437-3. doi: 
            10.1016/j.molcel.2022.05.003."
COMMENT     Sequence Label: pPlacPbuCas13b-gRNA-NT
FEATURES             Location/Qualifiers
     source          1..7798
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     139..158
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(225..327)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(719..736)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(853..1398)
                     /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(933..952)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        join(1510..1515,1516..1532,1533..1538)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     promoter        join(1688..1693,1694..1711,1712..1717)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     promoter        5117..5151
                     /label=J23119 promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119)"
     primer_bind     complement(5491..5510)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(join(7363..7798,1..224))
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
ORIGIN
        1 atacggaatt ccggatgagc attcatcagg cgggcaagaa tgtgaataaa ggccggataa
       61 aacttgtgct tatttttctt tacggtcttt aaaaaggccg taatatccag ctgaacggtc
      121 tggttatagg tacattgagc aactgactga aatgcctcaa aatgttcttt acgatgccat
      181 tgggatatat caacggtggt atatccagtg atttttttct ccattttagc ttccttagct
      241 cctgaaaatc tcgataactc aaaaaatacg cccggtagtg atcttatttc attatggtga
      301 aagttggaac ctcttacgtg ccgatcaacg tctcattttc gccaaaagtt ggcccagggc
      361 ttcccggtat caacagggac accaggattt atttattctg cgaagtgatc ttccgtcaca
      421 ggtatttatt cggcgcaaag tgcgtcgggt gatgctgcca acttactgat ttagtgtatg
      481 atggtgtttt tgaggtgctc cagtggcttc tgtttctatc agctgtccct cctgttcagc
      541 tactgacggg gtggtgcgta acggcaaaag caccgccgga catcagcgct agcggagtgt
      601 atactggctt actatgttgg cactgatgag ggtgtcagtg aagtgcttca tgtggcagga
      661 gaaaaaaggc tgcaccggtg cgtcagcaga atatgtgata caggatatat tccgcttcct
      721 cgctcactga ctcgctacgc tcggtcgttc gactgcggcg agcggaaatg gcttacgaac
      781 ggggcggaga tttcctggaa gatgccagga agatacttaa cagggaagtg agagggccgc
      841 ggcaaagccg tttttccata ggctccgccc ccctgacaag catcacgaaa tctgacgctc
      901 aaatcagtgg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggcgg
      961 ctccctcgtg cgctctcctg ttcctgcctt tcggtttacc ggtgtcattc cgctgttatg
     1021 gccgcgtttg tctcattcca cgcctgacac tcagttccgg gtaggcagtt cgctccaagc
     1081 tggactgtat gcacgaaccc cccgttcagt ccgaccgctg cgccttatcc ggtaactatc
     1141 gtcttgagtc caacccggaa agacatgcaa aagcaccact ggcagcagcc actggtaatt
     1201 gatttagagg agttagtctt gaagtcatgc gccggttaag gctaaactga aaggacaagt
     1261 tttggtgact gcgctcctcc aagccagtta cctcggttca aagagttggt agctcagaga
     1321 accttcgaaa aaccgccctg caaggcggtt ttttcgtttt cagagcaaga gattacgcgc
     1381 agaccaaaac gatctcaaga agatcatctt attaatcaga taaaatattt ctagatttca
     1441 gtgcaattta tctcttcaaa tgtagcacct gaagtcagcc ccatacgata taagttgtaa
     1501 ttctcatgtt tgacagctta tcatcgataa gctttaatgc ggtagtttat cacagttaaa
     1561 ttgctaacgc agtcaggcac cgtgtatgaa atctaacaat gcgctcatcg tcatcctcgg
     1621 caccgtcacc ctggatgctg taggcatagg cttggttatg ccggtactgc cgggcctctt
     1681 gcgggatttt acactttatg cttccggctc gtatgttagg aggtctttat catgcaaaaa
     1741 caagataaat tatttgtaga tagaaagaaa aatgctatat ttgcctttcc taaatatata
     1801 acaataatgg aaaacaaaga gaaacctgaa ccaatttatt atgaattgac agataagcat
     1861 ttctgggctg cctttcttaa cttggcgcgc cacaatgtct ataccaccat taatcacatc
     1921 aacagacgtt tggaaatagc agagctaaaa gacgatggat atatgatggg cattaaaggc
     1981 agctggaacg agcaagcgaa aaaactcgac aaaaaagttc ggctgagaga cttgatcatg
     2041 aaacactttc cttttctgga agctgcagct tatgaaatga caaactctaa aagtcccaac
     2101 aataaggagc aacgggaaaa ggaacaatcc gaagccctat cacttaacaa cttgaagaat
     2161 gttctattca ttttcttgga aaaacttcag gttctcagga actactactc ccattacaag
     2221 tattctgaag aatccccaaa accgatattt gaaacaagcc ttttgaaaaa catgtataag
     2281 gttttcgatg ccaatgtgcg gttagtaaag agagactata tgcatcatga gaatatagac
     2341 atgcaaagag attttacaca tctcaacagg aaaaaacaag tgggacgaac aaaaaacatc
     2401 atagacagtc ctaactttca ttaccatttt gcagataaag aaggcaatat gaccatagca
     2461 ggcttactgt tcttcgtctc tttatttctt gataaaaaag acgctatttg gatgcagaaa
     2521 aagctaaagg gatttaaaga tggcagaaac ttgcgtgagc aaatgacaaa cgaagtattt
     2581 tgcagaagcc gtatatcttt gcctaagcta aaattagaga atgtacagac aaaagactgg
     2641 atgcaacttg atatgcttaa cgaattggtt cgctgtccca aatcgctgta tgaacggtta
     2701 cgagaaaaag atagggaaag ttttaaagta cctttcgata ttttctctga cgactataat
     2761 gctgaagaag aaccatttaa gaacacgctt gtaaggcatc aagaccgctt tccatatttt
     2821 gtcttgagat atttcgattt aaatgagata ttcgaacaac tccgcttcca aatagattta
     2881 ggtacctatc atttctctat ctataacaaa cgtatcggtg acgaagatga agttcgccat
     2941 ctcacacatc atctatatgg ctttgcgcgc attcaggact ttgcaccaca aaaccaacca
     3001 gaagaatgga gaaaattagt gaaagactta gatcattttg agacatcgca agagccttac
     3061 atatctaaga ctgctccaca ttaccattta gagaacgaaa agataggcat taagttttgc
     3121 agtgcccaca acaacctctt tccatctctt cagaccgaca aaacatgcaa tggaagaagt
     3181 aaatttaatc tcggaacaca atttacagca gaagcatttt taagtgtcca cgaattgttg
     3241 ccgatgatgt tttattatct gttattgacc aaggattact ccagaaaaga aagcgcagac
     3301 aaagtagagg gaattataag aaaagaaata agcaacatat acgctatcta cgatgctttt
     3361 gccaataatg agataaatag cattgcagac ttaacccgtc ggcttcaaaa tacaaatata
     3421 ctacaaggcc atttgccaaa acaaatgatt tccatattga aaggcagaca aaaagatatg
     3481 ggaaaagagg ctgagcgcaa aattggggaa atgatagacg acacccaaag aagacttgac
     3541 ttgctctgca agcagacaaa tcagaagata aggattggca aaaggaatgc cggtctgcta
     3601 aaatcgggaa aaattgcaga ttggttggta aatgacatga tgcgcttcca acctgttcaa
     3661 aaagaccaaa ataacattcc aataaacaat agcaaggcta acagcaccga atatcgaatg
     3721 cttcagcgtg cgcttgccct tttcggcagt gagaacttcc gccttaaggc atattttaat
     3781 caaatgaatc ttgtggggaa tgacaatccg catcctttcc ttgcagaaac acaatgggaa
     3841 catcaaacta acattctgtc tttctaccga aactatttgg aagcgaggaa aaagtatctt
     3901 aagggcttaa aaccacaaaa ctggaaacaa tatcagcatt ttctcatact taaagtacag
     3961 aagaccaacc gcaacacatt agttacagga tggaaaaaca gtttcaatct cccccgtgga
     4021 atatttaccc aaccaataag ggaatggttt gagaaacata ataattcaaa aagaatatac
     4081 gaccaaatac tatcttttga ccgagtaggg tttgttgcaa aagctattcc cttatacttt
     4141 gcagaagaat ataaagacaa tgtacagcct ttctacgatt atccgttcaa tatcggcaac
     4201 agacttaaac caaaaaaaag gcaattctta gataaaaaag agcgtgtcga attgtggcaa
     4261 aagaataaag agctattcaa aaattatccg tctgaaaaga aaaagacaga cttagcttat
     4321 cttgattttt tatcatggaa aaagttcgaa cgcgaactgc gacttatcaa gaatcaggat
     4381 atcgtcacat ggttgatgtt caaagaacta ttcaatatgg ctactgtgga aggattgaag
     4441 attggagaaa tacatctgag ggacatcgat acaaacacgg caaatgagga aagcaacaat
     4501 attctaaata gaatcatgcc catgaagttg cctgttaaaa catacgaaac agacaataaa
     4561 ggaaatatat taaaggaaag acctttagca accttttata tcgaagaaac ggaaaccaaa
     4621 gtcctgaaac agggcaattt caaagcattg gtaaaagacc gaagactcaa tgggctattc
     4681 tcatttgcag aaactacaga cttgaactta gaggaacatc ctatcagcaa attgagtgta
     4741 gaccttgagc ttataaaata tcagactaca aggattagta tctttgagat gacattagga
     4801 ctggaaaaga aactcataga caaatattcc acactaccta cagatagctt cagaaatatg
     4861 cttgaaaggt ggttgcagtg caaagctaat agaccggaat taaagaatta cgttaatagt
     4921 ttgattgcag tgagaaatgc tttttcccac aaccagtatc ccatgtatga tgccacactc
     4981 tttgctgaag ttaaaaaatt tactcttttc ccttctgtcg atacaaaaaa gattgagcta
     5041 aatatagcgc cacagttact ggaaatagta ggaaaagcca taaaagaaat tgaaaaatca
     5101 gaaaataaaa actaaattga cagctagctc agtcctaggt ataatgctag cgctgagttg
     5161 catctgcctt ctttttgaaa ggtaaaaaca acgtaccttc gtacgttgca tctgccttct
     5221 ttttgaaagg taaaaacaac atcgtccatt ccgacagcat cgccagtcac tatggcgtgc
     5281 tgctagcgct atatgcgttg atgcaatttc tatgcgcacc cgttctcgga gcactgtccg
     5341 accgctttgg ccgccgccca gtcctgctcg cttcgctact tggagccact atcgactacg
     5401 cgatcatggc gaccacaccc gtcctgtgga tcctctacgc cggacgcatc gtggccggca
     5461 tcaccggcgc cacaggtgcg gttgctggcg cctatatcgc cgacatcacc gatggggaag
     5521 atcgggctcg ccacttcggg ctcatgagcg cttgtttcgg cgtgggtatg gtggcaggcc
     5581 ccgtggccgg gggactgttg ggcgccatct ccttgcatgc accattcctt gcggcggcgg
     5641 tgctcaacgg cctcaaccta ctactgggct gcttcctaat gcaggagtcg cataagggag
     5701 agcgtcgacc gatgcccttg agagccttca acccagtcag ctccttccgg tgggcgcggg
     5761 gcatgactat cgtcgccgca cttatgactg tcttctttat catgcaactc gtaggacagg
     5821 tgccggcagc gctctgggtc attttcggcg aggaccgctt tcgctggagc gcgacgatga
     5881 tcggcctgtc gcttgcggta ttcggaatct tgcacgccct cgctcaagcc ttcgtcactg
     5941 gtcccgccac caaacgtttc ggcgagaagc aggccattat cgccggcatg gcggccgacg
     6001 cgctgggcta cgtcttgctg gcgttcgcga cgcgaggctg gatggccttc cccattatga
     6061 ttcttctcgc ttccggcggc atcgggatgc ccgcgttgca ggccatgctg tccaggcagg
     6121 tagatgacga ccatcaggga cagcttcaag gatcgctcgc ggctcttacc agcctaactt
     6181 cgatcattgg accgctgatc gtcacggcga tttatgccgc ctcggcgagc acatggaacg
     6241 ggttggcatg gattgtaggc gccgccctat accttgtctg cctccccgcg ttgcgtcgcg
     6301 gtgcatggag ccgggccacc tcgacctgaa tggaagccgg cggcacctcg ctaacggatt
     6361 caccactcca agaattggag ccaatcaatt cttgcggaga actgtgaatg cgcaaaccaa
     6421 cccttggcag aacatatcca tcgcgtccgc catctccagc agccgcacgc ggcgcatctc
     6481 gggcagcgtt gggtcctggc cacgggtgcg catgatcgtg ctcctgtcgt tgaggacccg
     6541 gctaggctgg cggggttgcc ttactggtta gcagaatgaa tcaccgatac gcgagcgaac
     6601 gtgaagcgac tgctgctgca aaacgtctgc gacctgagca acaacatgaa tggtcttcgg
     6661 tttccgtgtt tcgtaaagtc tggaaacgcg gaagtcccct acgtgctgct gaagttgccc
     6721 gcaacagaga gtggaaccaa ccggtgatac cacgatacta tgactgagag tcaacgccat
     6781 gagcggcctc atttcttatt ctgagttaca acagtccgca ccgctgccgg tagctccttc
     6841 cggtgggcgc ggggcatgac tatcgtcgcc gcacttatga ctgtcttctt tatcatgcaa
     6901 ctcgtaggac aggtgccggc agcgcccaac agtcccccgg ccacggggcc tgccaccata
     6961 cccacgccga aacaagcgcc ctgcaccatt atgttccgga tctgcatcgc aggatgctgc
     7021 tggctaccct gtggaacacc tacatctgta ttaacgaagc gctaaccgtt tttatcaggc
     7081 tctgggaggc agaataaatg atcatatcgt caattattac ctccacgggg agagcctgag
     7141 caaactggcc tcaggcattt gagaagcaca cggtcacact gcttccggta gtcaataaac
     7201 cggtaaacca gcaatagaca taagcggcta tttaacgacc ctgccctgaa ccgacgaccg
     7261 ggtcgaattt gctttcgaat ttctgccatt catccgctta ttatcactta ttcaggcgta
     7321 gcaaccaggc gtttaagggc accaataact gccttaaaaa aattacgccc cgccctgcca
     7381 ctcatcgcag tactgttgta attcattaag cattctgccg acatggaagc catcacaaac
     7441 ggcatgatga acctgaatcg ccagcggcat cagcaccttg tcgccttgcg tataatattt
     7501 gcccatggtg aaaacggggg cgaagaagtt gtccatattg gccacgttta aatcaaaact
     7561 ggtgaaactc acccagggat tggctgagac gaaaaacata ttctcaataa accctttagg
     7621 gaaataggcc aggttttcac cgtaacacgc cacatcttgc gaatatatgt gtagaaactg
     7681 ccggaaatcg tcgtggtatt cactccagag cgatgaaaac gtttcagttt gctcatggaa
     7741 aacggtgtaa caagggtgaa cactatccca tatcaccagc tcaccgtctt tcattgcc
//
