LOCUS       sequence-374002-        4360 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Constitutive expression of single-spacer CRISPR array with spacer #1
            targeting deGFP mRNA 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-1
FEATURES             Location/Qualifiers
     source          1..4360
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(288..947)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     862..881
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(948..1050)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(1442..1459)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1576..2121)
                     /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(1656..1675)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        join(2233..2238,2239..2255,2256..2261)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     promoter        2384..2418
                     /label=J23119 promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119)"
     primer_bind     2460..2481
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2776..2795)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 caggctctgg gaggcagaat aaatgatcat atcgtcaatt attacctcca cggggagagc
       61 ctgagcaaac tggcctcagg catttgagaa gcacacggtc acactgcttc cggtagtcaa
      121 taaaccggta aaccagcaat agacataagc ggctatttaa cgaccctgcc ctgaaccgac
      181 gaccgggtcg aatttgcttt cgaatttctg ccattcatcc gcttattatc acttattcag
      241 gcgtagcaac caggcgttta agggcaccaa taactgcctt aaaaaaatta cgccccgccc
      301 tgccactcat cgcagtactg ttgtaattca ttaagcattc tgccgacatg gaagccatca
      361 caaacggcat gatgaacctg aatcgccagc ggcatcagca ccttgtcgcc ttgcgtataa
      421 tatttgccca tggtgaaaac gggggcgaag aagttgtcca tattggccac gtttaaatca
      481 aaactggtga aactcaccca gggattggct gagacgaaaa acatattctc aataaaccct
      541 ttagggaaat aggccaggtt ttcaccgtaa cacgccacat cttgcgaata tatgtgtaga
      601 aactgccgga aatcgtcgtg gtattcactc cagagcgatg aaaacgtttc agtttgctca
      661 tggaaaacgg tgtaacaagg gtgaacacta tcccatatca ccagctcacc gtctttcatt
      721 gccatacgga attccggatg agcattcatc aggcgggcaa gaatgtgaat aaaggccgga
      781 taaaacttgt gcttattttt ctttacggtc tttaaaaagg ccgtaatatc cagctgaacg
      841 gtctggttat aggtacattg agcaactgac tgaaatgcct caaaatgttc tttacgatgc
      901 cattgggata tatcaacggt ggtatatcca gtgatttttt tctccatttt agcttcctta
      961 gctcctgaaa atctcgataa ctcaaaaaat acgcccggta gtgatcttat ttcattatgg
     1021 tgaaagttgg aacctcttac gtgccgatca acgtctcatt ttcgccaaaa gttggcccag
     1081 ggcttcccgg tatcaacagg gacaccagga tttatttatt ctgcgaagtg atcttccgtc
     1141 acaggtattt attcggcgca aagtgcgtcg ggtgatgctg ccaacttact gatttagtgt
     1201 atgatggtgt ttttgaggtg ctccagtggc ttctgtttct atcagctgtc cctcctgttc
     1261 agctactgac ggggtggtgc gtaacggcaa aagcaccgcc ggacatcagc gctagcggag
     1321 tgtatactgg cttactatgt tggcactgat gagggtgtca gtgaagtgct tcatgtggca
     1381 ggagaaaaaa ggctgcaccg gtgcgtcagc agaatatgtg atacaggata tattccgctt
     1441 cctcgctcac tgactcgcta cgctcggtcg ttcgactgcg gcgagcggaa atggcttacg
     1501 aacggggcgg agatttcctg gaagatgcca ggaagatact taacagggaa gtgagagggc
     1561 cgcggcaaag ccgtttttcc ataggctccg cccccctgac aagcatcacg aaatctgacg
     1621 ctcaaatcag tggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     1681 cggctccctc gtgcgctctc ctgttcctgc ctttcggttt accggtgtca ttccgctgtt
     1741 atggccgcgt ttgtctcatt ccacgcctga cactcagttc cgggtaggca gttcgctcca
     1801 agctggactg tatgcacgaa ccccccgttc agtccgaccg ctgcgcctta tccggtaact
     1861 atcgtcttga gtccaacccg gaaagacatg caaaagcacc actggcagca gccactggta
     1921 attgatttag aggagttagt cttgaagtca tgcgccggtt aaggctaaac tgaaaggaca
     1981 agttttggtg actgcgctcc tccaagccag ttacctcggt tcaaagagtt ggtagctcag
     2041 agaaccttcg aaaaaccgcc ctgcaaggcg gttttttcgt tttcagagca agagattacg
     2101 cgcagaccaa aacgatctca agaagatcat cttattaatc agataaaata tttctagatt
     2161 tcagtgcaat ttatctcttc aaatgtagca cctgaagtca gccccatacg atataagttg
     2221 taattctcat gtttgacagc ttatcatcga taagctttaa tgcggtagtt tatcacagtt
     2281 aaattgctaa cgcagtcagg caccgtgtat gaaatctaac aatgcgctca tcgtcatcct
     2341 cggcaccgtc accctggatg ctgtaggcat aggcttggtt taattgacag ctagctcagt
     2401 cctaggtata atgctagcgc tgagttgcat ctgccttctt tttgaaaggt aaaaacaacc
     2461 gtcgccgtcc agctcgacca ggatgggaag ttgcatctgc cttctttttg aaaggtaaaa
     2521 acaacatcgt ccattccgac agcatcgcca gtcactatgg cgtgctgcta gcgctatatg
     2581 cgttgatgca atttctatgc gcacccgttc tcggagcact gtccgaccgc tttggccgcc
     2641 gcccagtcct gctcgcttcg ctacttggag ccactatcga ctacgcgatc atggcgacca
     2701 cacccgtcct gtggatcctc tacgccggac gcatcgtggc cggcatcacc ggcgccacag
     2761 gtgcggttgc tggcgcctat atcgccgaca tcaccgatgg ggaagatcgg gctcgccact
     2821 tcgggctcat gagcgcttgt ttcggcgtgg gtatggtggc aggccccgtg gccgggggac
     2881 tgttgggcgc catctccttg catgcaccat tccttgcggc ggcggtgctc aacggcctca
     2941 acctactact gggctgcttc ctaatgcagg agtcgcataa gggagagcgt cgaccgatgc
     3001 ccttgagagc cttcaaccca gtcagctcct tccggtgggc gcggggcatg actatcgtcg
     3061 ccgcacttat gactgtcttc tttatcatgc aactcgtagg acaggtgccg gcagcgctct
     3121 gggtcatttt cggcgaggac cgctttcgct ggagcgcgac gatgatcggc ctgtcgcttg
     3181 cggtattcgg aatcttgcac gccctcgctc aagccttcgt cactggtccc gccaccaaac
     3241 gtttcggcga gaagcaggcc attatcgccg gcatggcggc cgacgcgctg ggctacgtct
     3301 tgctggcgtt cgcgacgcga ggctggatgg ccttccccat tatgattctt ctcgcttccg
     3361 gcggcatcgg gatgcccgcg ttgcaggcca tgctgtccag gcaggtagat gacgaccatc
     3421 agggacagct tcaaggatcg ctcgcggctc ttaccagcct aacttcgatc attggaccgc
     3481 tgatcgtcac ggcgatttat gccgcctcgg cgagcacatg gaacgggttg gcatggattg
     3541 taggcgccgc cctatacctt gtctgcctcc ccgcgttgcg tcgcggtgca tggagccggg
     3601 ccacctcgac ctgaatggaa gccggcggca cctcgctaac ggattcacca ctccaagaat
     3661 tggagccaat caattcttgc ggagaactgt gaatgcgcaa accaaccctt ggcagaacat
     3721 atccatcgcg tccgccatct ccagcagccg cacgcggcgc atctcgggca gcgttgggtc
     3781 ctggccacgg gtgcgcatga tcgtgctcct gtcgttgagg acccggctag gctggcgggg
     3841 ttgccttact ggttagcaga atgaatcacc gatacgcgag cgaacgtgaa gcgactgctg
     3901 ctgcaaaacg tctgcgacct gagcaacaac atgaatggtc ttcggtttcc gtgtttcgta
     3961 aagtctggaa acgcggaagt cccctacgtg ctgctgaagt tgcccgcaac agagagtgga
     4021 accaaccggt gataccacga tactatgact gagagtcaac gccatgagcg gcctcatttc
     4081 ttattctgag ttacaacagt ccgcaccgct gccggtagct ccttccggtg ggcgcggggc
     4141 atgactatcg tcgccgcact tatgactgtc ttctttatca tgcaactcgt aggacaggtg
     4201 ccggcagcgc ccaacagtcc cccggccacg gggcctgcca ccatacccac gccgaaacaa
     4261 gcgccctgca ccattatgtt ccggatctgc atcgcaggat gctgctggct accctgtgga
     4321 acacctacat ctgtattaac gaagcgctaa ccgtttttat
//
