LOCUS       sequence-374001-        4360 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Constitutive expression of single-spacer CRISPR array with 
            non-targeting spacer for PbuCas13b in bacteria..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4360)
  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 4360)
  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: pPbuCas13b-gRNA-NT
FEATURES             Location/Qualifiers
     source          1..4360
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     242..261
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(328..430)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(822..839)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(956..1501)
                     /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(1036..1055)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        join(1613..1618,1619..1635,1636..1641)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     promoter        1764..1798
                     /label=J23119 promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119)"
     primer_bind     complement(2156..2175)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(join(4028..4360,1..327))
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
ORIGIN
        1 gtattcactc cagagcgatg aaaacgtttc agtttgctca tggaaaacgg tgtaacaagg
       61 gtgaacacta tcccatatca ccagctcacc gtctttcatt gccatacgga attccggatg
      121 agcattcatc aggcgggcaa gaatgtgaat aaaggccgga taaaacttgt gcttattttt
      181 ctttacggtc tttaaaaagg ccgtaatatc cagctgaacg gtctggttat aggtacattg
      241 agcaactgac tgaaatgcct caaaatgttc tttacgatgc cattgggata tatcaacggt
      301 ggtatatcca gtgatttttt tctccatttt agcttcctta gctcctgaaa atctcgataa
      361 ctcaaaaaat acgcccggta gtgatcttat ttcattatgg tgaaagttgg aacctcttac
      421 gtgccgatca acgtctcatt ttcgccaaaa gttggcccag ggcttcccgg tatcaacagg
      481 gacaccagga tttatttatt ctgcgaagtg atcttccgtc acaggtattt attcggcgca
      541 aagtgcgtcg ggtgatgctg ccaacttact gatttagtgt atgatggtgt ttttgaggtg
      601 ctccagtggc ttctgtttct atcagctgtc cctcctgttc agctactgac ggggtggtgc
      661 gtaacggcaa aagcaccgcc ggacatcagc gctagcggag tgtatactgg cttactatgt
      721 tggcactgat gagggtgtca gtgaagtgct tcatgtggca ggagaaaaaa ggctgcaccg
      781 gtgcgtcagc agaatatgtg atacaggata tattccgctt cctcgctcac tgactcgcta
      841 cgctcggtcg ttcgactgcg gcgagcggaa atggcttacg aacggggcgg agatttcctg
      901 gaagatgcca ggaagatact taacagggaa gtgagagggc cgcggcaaag ccgtttttcc
      961 ataggctccg cccccctgac aagcatcacg aaatctgacg ctcaaatcag tggtggcgaa
     1021 acccgacagg actataaaga taccaggcgt ttccccctgg cggctccctc gtgcgctctc
     1081 ctgttcctgc ctttcggttt accggtgtca ttccgctgtt atggccgcgt ttgtctcatt
     1141 ccacgcctga cactcagttc cgggtaggca gttcgctcca agctggactg tatgcacgaa
     1201 ccccccgttc agtccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     1261 gaaagacatg caaaagcacc actggcagca gccactggta attgatttag aggagttagt
     1321 cttgaagtca tgcgccggtt aaggctaaac tgaaaggaca agttttggtg actgcgctcc
     1381 tccaagccag ttacctcggt tcaaagagtt ggtagctcag agaaccttcg aaaaaccgcc
     1441 ctgcaaggcg gttttttcgt tttcagagca agagattacg cgcagaccaa aacgatctca
     1501 agaagatcat cttattaatc agataaaata tttctagatt tcagtgcaat ttatctcttc
     1561 aaatgtagca cctgaagtca gccccatacg atataagttg taattctcat gtttgacagc
     1621 ttatcatcga taagctttaa tgcggtagtt tatcacagtt aaattgctaa cgcagtcagg
     1681 caccgtgtat gaaatctaac aatgcgctca tcgtcatcct cggcaccgtc accctggatg
     1741 ctgtaggcat aggcttggtt taattgacag ctagctcagt cctaggtata atgctagcgc
     1801 tgagttgcat ctgccttctt tttgaaaggt aaaaacaacc tcgtatgttg tgtggaattg
     1861 tgagcggatg ttgcatctgc cttctttttg aaaggtaaaa acaacatcgt ccattccgac
     1921 agcatcgcca gtcactatgg cgtgctgcta gcgctatatg cgttgatgca atttctatgc
     1981 gcacccgttc tcggagcact gtccgaccgc tttggccgcc gcccagtcct gctcgcttcg
     2041 ctacttggag ccactatcga ctacgcgatc atggcgacca cacccgtcct gtggatcctc
     2101 tacgccggac gcatcgtggc cggcatcacc ggcgccacag gtgcggttgc tggcgcctat
     2161 atcgccgaca tcaccgatgg ggaagatcgg gctcgccact tcgggctcat gagcgcttgt
     2221 ttcggcgtgg gtatggtggc aggccccgtg gccgggggac tgttgggcgc catctccttg
     2281 catgcaccat tccttgcggc ggcggtgctc aacggcctca acctactact gggctgcttc
     2341 ctaatgcagg agtcgcataa gggagagcgt cgaccgatgc ccttgagagc cttcaaccca
     2401 gtcagctcct tccggtgggc gcggggcatg actatcgtcg ccgcacttat gactgtcttc
     2461 tttatcatgc aactcgtagg acaggtgccg gcagcgctct gggtcatttt cggcgaggac
     2521 cgctttcgct ggagcgcgac gatgatcggc ctgtcgcttg cggtattcgg aatcttgcac
     2581 gccctcgctc aagccttcgt cactggtccc gccaccaaac gtttcggcga gaagcaggcc
     2641 attatcgccg gcatggcggc cgacgcgctg ggctacgtct tgctggcgtt cgcgacgcga
     2701 ggctggatgg ccttccccat tatgattctt ctcgcttccg gcggcatcgg gatgcccgcg
     2761 ttgcaggcca tgctgtccag gcaggtagat gacgaccatc agggacagct tcaaggatcg
     2821 ctcgcggctc ttaccagcct aacttcgatc attggaccgc tgatcgtcac ggcgatttat
     2881 gccgcctcgg cgagcacatg gaacgggttg gcatggattg taggcgccgc cctatacctt
     2941 gtctgcctcc ccgcgttgcg tcgcggtgca tggagccggg ccacctcgac ctgaatggaa
     3001 gccggcggca cctcgctaac ggattcacca ctccaagaat tggagccaat caattcttgc
     3061 ggagaactgt gaatgcgcaa accaaccctt ggcagaacat atccatcgcg tccgccatct
     3121 ccagcagccg cacgcggcgc atctcgggca gcgttgggtc ctggccacgg gtgcgcatga
     3181 tcgtgctcct gtcgttgagg acccggctag gctggcgggg ttgccttact ggttagcaga
     3241 atgaatcacc gatacgcgag cgaacgtgaa gcgactgctg ctgcaaaacg tctgcgacct
     3301 gagcaacaac atgaatggtc ttcggtttcc gtgtttcgta aagtctggaa acgcggaagt
     3361 cccctacgtg ctgctgaagt tgcccgcaac agagagtgga accaaccggt gataccacga
     3421 tactatgact gagagtcaac gccatgagcg gcctcatttc ttattctgag ttacaacagt
     3481 ccgcaccgct gccggtagct ccttccggtg ggcgcggggc atgactatcg tcgccgcact
     3541 tatgactgtc ttctttatca tgcaactcgt aggacaggtg ccggcagcgc ccaacagtcc
     3601 cccggccacg gggcctgcca ccatacccac gccgaaacaa gcgccctgca ccattatgtt
     3661 ccggatctgc atcgcaggat gctgctggct accctgtgga acacctacat ctgtattaac
     3721 gaagcgctaa ccgtttttat caggctctgg gaggcagaat aaatgatcat atcgtcaatt
     3781 attacctcca cggggagagc ctgagcaaac tggcctcagg catttgagaa gcacacggtc
     3841 acactgcttc cggtagtcaa taaaccggta aaccagcaat agacataagc ggctatttaa
     3901 cgaccctgcc ctgaaccgac gaccgggtcg aatttgcttt cgaatttctg ccattcatcc
     3961 gcttattatc acttattcag gcgtagcaac caggcgttta agggcaccaa taactgcctt
     4021 aaaaaaatta cgccccgccc tgccactcat cgcagtactg ttgtaattca ttaagcattc
     4081 tgccgacatg gaagccatca caaacggcat gatgaacctg aatcgccagc ggcatcagca
     4141 ccttgtcgcc ttgcgtataa tatttgccca tggtgaaaac gggggcgaag aagttgtcca
     4201 tattggccac gtttaaatca aaactggtga aactcaccca gggattggct gagacgaaaa
     4261 acatattctc aataaaccct ttagggaaat aggccaggtt ttcaccgtaa cacgccacat
     4321 cttgcgaata tatgtgtaga aactgccgga aatcgtcgtg
//
