LOCUS       sequence-178222-        8098 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression for bzCas13b, driven by the lac promoter, and 
            DR-spacer-DR sequence driven by js23119. New spacer sequences can be
            cloned in between the DRs by digesting the plasmid with BsaI..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8098)
  AUTHORS   Smargon AA, Cox DB, Pyzocha NK, Zheng K, Slaymaker IM, Gootenberg 
            JS, Abudayyeh OA, Essletzbichler P, Shmakov S, Makarova KS, Koonin 
            EV, Zhang F
  TITLE     Cas13b Is a Type VI-B CRISPR-Associated RNA-Guided RNase 
            Differentially Regulated by Accessory Proteins Csx27 and Csx28.
  JOURNAL   Mol Cell. 2017 Feb 16;65(4):618-630.e7. doi: 
            10.1016/j.molcel.2016.12.023. Epub 2017 Jan 5.
  PUBMED    28065598
REFERENCE   2  (bases 1 to 8098)
  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: 
            "10.1016/j.molcel.2016.12.023"; journalName: "Mol Cell"; date: 
            "2017-02-16- 16"; volume: "65"; issue: "4"; pages: "618-630.e7"
COMMENT     Sequence Label: pBZCas13b
FEATURES             Location/Qualifiers
     source          1..8098
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1303..1337
                     /label=J23119 promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119)"
     primer_bind     complement(1687..1706)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(3559..4218)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4133..4152
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(4219..4321)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(4713..4730)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4847..5392)
                     /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(4927..4946)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        join(5504..5509,5510..5526,5527..5532)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     promoter        join(5682..5687,5688..5705,5706..5711)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
ORIGIN
        1 tgtaaagagg agttgaaaaa tcttttgcct gaaaaagtat ttcagcattt accttttaag
       61 ttgggaggat attttcaaca aaaatatttg taccaatttt acacttgtta tttagataaa
      121 cggttggaat acatcagtgg gttggtgcaa caagccgaaa actttaaatc cgaaaataaa
      181 gtcttcaaaa aggtagaaaa tgagtgtttt aaatttttga aaaaacaaaa ttatactcac
      241 aaagaattag atgctcgggt tcagtctata ttggggtatc ctatcttttt agagcggggc
      301 tttatggatg aaaaacctac gataatcaaa ggaaaaactt ttaaaggaaa tgaagctctt
      361 tttgccgatt ggtttaggta ttacaaggaa tatcagaatt ttcagacatt ttatgatact
      421 gaaaattatc cgcttgtaga attggaaaaa aaacaagcag accgaaaacg aaaaactaaa
      481 atctaccaac aaaagaaaaa cgatgtgttt acacttttga tggcaaaaca tattttcaaa
      541 tcggtattca agcaggatag catcgatcag ttttcgttag aagatttgta tcaatctcgg
      601 gaggaacgct taggaaatca ggagagagcc agacaaacgg gcgagcgaaa tactaactat
      661 atttggaaca aaaccgttga tttaaagttg tgtgacggaa aaattacggt agaaaatgta
      721 aagctaaaaa atgtaggaga tttcataaaa tatgaatacg accaaagggt tcaagctttc
      781 ttaaaatatg aggaaaatat agaatggcag gcgtttctaa tcaaagagag taaggaagag
      841 gaaaattatc cgtatgtggt ggaaagagaa atagagcaat atgaaaaagt gagaagggaa
      901 gaactgttga aggaagtcca ccttatagag gaatatattt tggaaaaggt aaaagataag
      961 gaaatattaa aaaaagggga taatcaaaat ttcaaatact acattctcaa cggactttta
     1021 aagcaactta aaaatgaaga tgtggagagc tataaggtat tcaacttaaa tactgaacct
     1081 gaagatgtga acattaacca actaaaacaa gaagcaaccg atttagaaca aaaagctttt
     1141 gttttaacct atattcgtaa taaatttgca cacaaccaat tacctaaaaa agaattttgg
     1201 gactattgtc aagaaaaata cggcaaaata gaaaaagaaa aaacctatgc cgagtatttt
     1261 gctgaggtat ttaaaaagga aaaagaagca ctgataaagt aattgacagc tagctcagtc
     1321 ctaggtataa tgctagcgct gagttggaac tgctctcatt ttggagggta atcacaacga
     1381 gaccatggcg gaaggtctca gttggaactg ctctcatttt ggagggtaat cacaacatcg
     1441 tccattccga cagcatcgcc agtcactatg gcgtgctgct agcgctatat gcgttgatgc
     1501 aatttctatg cgcacccgtt ctcggagcac tgtccgaccg ctttggccgc cgcccagtcc
     1561 tgctcgcttc gctacttgga gccactatcg actacgcgat catggcgacc acacccgtcc
     1621 tgtggatcct ctacgccgga cgcatcgtgg ccggcatcac cggcgccaca ggtgcggttg
     1681 ctggcgccta tatcgccgac atcaccgatg gggaagatcg ggctcgccac ttcgggctca
     1741 tgagcgcttg tttcggcgtg ggtatggtgg caggccccgt ggccggggga ctgttgggcg
     1801 ccatctcctt gcatgcacca ttccttgcgg cggcggtgct caacggcctc aacctactac
     1861 tgggctgctt cctaatgcag gagtcgcata agggagagcg tcgaccgatg cccttgagag
     1921 ccttcaaccc agtcagctcc ttccggtggg cgcggggcat gactatcgtc gccgcactta
     1981 tgactgtctt ctttatcatg caactcgtag gacaggtgcc ggcagcgctc tgggtcattt
     2041 tcggcgagga ccgctttcgc tggagcgcga cgatgatcgg cctgtcgctt gcggtattcg
     2101 gaatcttgca cgccctcgct caagccttcg tcactggtcc cgccaccaaa cgtttcggcg
     2161 agaagcaggc cattatcgcc ggcatggcgg ccgacgcgct gggctacgtc ttgctggcgt
     2221 tcgcgacgcg aggctggatg gccttcccca ttatgattct tctcgcttcc ggcggcatcg
     2281 ggatgcccgc gttgcaggcc atgctgtcca ggcaggtaga tgacgaccat cagggacagc
     2341 ttcaaggatc gctcgcggct cttaccagcc taacttcgat cattggaccg ctgatcgtca
     2401 cggcgattta tgccgcctcg gcgagcacat ggaacgggtt ggcatggatt gtaggcgccg
     2461 ccctatacct tgtctgcctc cccgcgttgc gtcgcggtgc atggagccgg gccacctcga
     2521 cctgaatgga agccggcggc acctcgctaa cggattcacc actccaagaa ttggagccaa
     2581 tcaattcttg cggagaactg tgaatgcgca aaccaaccct tggcagaaca tatccatcgc
     2641 gtccgccatc tccagcagcc gcacgcggcg catctcgggc agcgttgggt cctggccacg
     2701 ggtgcgcatg atcgtgctcc tgtcgttgag gacccggcta ggctggcggg gttgccttac
     2761 tggttagcag aatgaatcac cgatacgcga gcgaacgtga agcgactgct gctgcaaaac
     2821 gtctgcgacc tgagcaacaa catgaatggt cttcggtttc cgtgtttcgt aaagtctgga
     2881 aacgcggaag tcccctacgt gctgctgaag ttgcccgcaa cagagagtgg aaccaaccgg
     2941 tgataccacg atactatgac tgagagtcaa cgccatgagc ggcctcattt cttattctga
     3001 gttacaacag tccgcaccgc tgccggtagc tccttccggt gggcgcgggg catgactatc
     3061 gtcgccgcac ttatgactgt cttctttatc atgcaactcg taggacaggt gccggcagcg
     3121 cccaacagtc ccccggccac ggggcctgcc accataccca cgccgaaaca agcgccctgc
     3181 accattatgt tccggatctg catcgcagga tgctgctggc taccctgtgg aacacctaca
     3241 tctgtattaa cgaagcgcta accgttttta tcaggctctg ggaggcagaa taaatgatca
     3301 tatcgtcaat tattacctcc acggggagag cctgagcaaa ctggcctcag gcatttgaga
     3361 agcacacggt cacactgctt ccggtagtca ataaaccggt aaaccagcaa tagacataag
     3421 cggctattta acgaccctgc cctgaaccga cgaccgggtc gaatttgctt tcgaatttct
     3481 gccattcatc cgcttattat cacttattca ggcgtagcaa ccaggcgttt aagggcacca
     3541 ataactgcct taaaaaaatt acgccccgcc ctgccactca tcgcagtact gttgtaattc
     3601 attaagcatt ctgccgacat ggaagccatc acaaacggca tgatgaacct gaatcgccag
     3661 cggcatcagc accttgtcgc cttgcgtata atatttgccc atggtgaaaa cgggggcgaa
     3721 gaagttgtcc atattggcca cgtttaaatc aaaactggtg aaactcaccc agggattggc
     3781 tgagacgaaa aacatattct caataaaccc tttagggaaa taggccaggt tttcaccgta
     3841 acacgccaca tcttgcgaat atatgtgtag aaactgccgg aaatcgtcgt ggtattcact
     3901 ccagagcgat gaaaacgttt cagtttgctc atggaaaacg gtgtaacaag ggtgaacact
     3961 atcccatatc accagctcac cgtctttcat tgccatacgg aattccggat gagcattcat
     4021 caggcgggca agaatgtgaa taaaggccgg ataaaacttg tgcttatttt tctttacggt
     4081 ctttaaaaag gccgtaatat ccagctgaac ggtctggtta taggtacatt gagcaactga
     4141 ctgaaatgcc tcaaaatgtt ctttacgatg ccattgggat atatcaacgg tggtatatcc
     4201 agtgattttt ttctccattt tagcttcctt agctcctgaa aatctcgata actcaaaaaa
     4261 tacgcccggt agtgatctta tttcattatg gtgaaagttg gaacctctta cgtgccgatc
     4321 aacgtctcat tttcgccaaa agttggccca gggcttcccg gtatcaacag ggacaccagg
     4381 atttatttat tctgcgaagt gatcttccgt cacaggtatt tattcggcgc aaagtgcgtc
     4441 gggtgatgct gccaacttac tgatttagtg tatgatggtg tttttgaggt gctccagtgg
     4501 cttctgtttc tatcagctgt ccctcctgtt cagctactga cggggtggtg cgtaacggca
     4561 aaagcaccgc cggacatcag cgctagcgga gtgtatactg gcttactatg ttggcactga
     4621 tgagggtgtc agtgaagtgc ttcatgtggc aggagaaaaa aggctgcacc ggtgcgtcag
     4681 cagaatatgt gatacaggat atattccgct tcctcgctca ctgactcgct acgctcggtc
     4741 gttcgactgc ggcgagcgga aatggcttac gaacggggcg gagatttcct ggaagatgcc
     4801 aggaagatac ttaacaggga agtgagaggg ccgcggcaaa gccgtttttc cataggctcc
     4861 gcccccctga caagcatcac gaaatctgac gctcaaatca gtggtggcga aacccgacag
     4921 gactataaag ataccaggcg tttccccctg gcggctccct cgtgcgctct cctgttcctg
     4981 cctttcggtt taccggtgtc attccgctgt tatggccgcg tttgtctcat tccacgcctg
     5041 acactcagtt ccgggtaggc agttcgctcc aagctggact gtatgcacga accccccgtt
     5101 cagtccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggaaagacat
     5161 gcaaaagcac cactggcagc agccactggt aattgattta gaggagttag tcttgaagtc
     5221 atgcgccggt taaggctaaa ctgaaaggac aagttttggt gactgcgctc ctccaagcca
     5281 gttacctcgg ttcaaagagt tggtagctca gagaaccttc gaaaaaccgc cctgcaaggc
     5341 ggttttttcg ttttcagagc aagagattac gcgcagacca aaacgatctc aagaagatca
     5401 tcttattaat cagataaaat atttctagat ttcagtgcaa tttatctctt caaatgtagc
     5461 acctgaagtc agccccatac gatataagtt gtaattctca tgtttgacag cttatcatcg
     5521 ataagcttta atgcggtagt ttatcacagt taaattgcta acgcagtcag gcaccgtgta
     5581 tgaaatctaa caatgcgctc atcgtcatcc tcggcaccgt caccctggat gctgtaggca
     5641 taggcttggt tatgccggta ctgccgggcc tcttgcggga ttttacactt tatgcttccg
     5701 gctcgtatgt taggaggtct ttatcatgga aaacaaaaca tctttgggga ataatattta
     5761 ttacaaccct tttaaaccac aagataaatc gtattttgcg ggttatttca atgcagcaat
     5821 ggaaaataca gatagtgtat ttagagaatt gggaaaaaga ttaaaaggta aagagtacac
     5881 ttcggagaat ttttttgatg ctatctttaa ggaaaatatt tcattagtgg aatacgaaag
     5941 gtatgtgaaa ttgctttctg attatttccc tatggctcgt cttttggata aaaaagaagt
     6001 tcctataaaa gaacgaaaag aaaattttaa aaagaatttt aaaggcatta taaaagccgt
     6061 aagagattta agaaactttt atacgcataa ggaacacgga gaggtagaaa ttacagatga
     6121 aatatttggc gtattggatg agatgctgaa aagcacggtt ttaacggtaa aaaagaaaaa
     6181 agtaaaaacc gataaaacaa aagaaatcct gaaaaaaagc atagaaaaac aattggatat
     6241 tttatgccaa aagaaattag aatatctaag agacacagca agaaaaatag aggagaagcg
     6301 tagaaatcaa agagaaagag gagagaaaga attggtagct ccattcaaat acagtgataa
     6361 aagggatgat ttaatagcag cgatttataa tgatgctttt gatgtttata tcgataagaa
     6421 aaaagatagc ctaaaagaat ccagcaaagc aaagtacaat accaaaagtg atcctcagca
     6481 agaagaaggc gatttgaaaa ttccaatttc gaaaaacgga gtagtcttcc tgctttcttt
     6541 gtttctcacc aaacaagaaa tacacgcatt caaatctaaa atagcagggt ttaaggctac
     6601 ggttatcgat gaagcaacgg tatctgaagc aacggtgagc catggaaaaa acagcatttg
     6661 ttttatggct acgcacgaaa ttttctcgca tttggcttac aaaaaattga aacgaaaagt
     6721 gagaacggct gaaatcaact atggtgaggc agaaaatgct gaacagcttt ccgtttatgc
     6781 caaagaaact ctaatgatgc agatgcttga tgaattgagc aaagtacccg atgtggtcta
     6841 tcagaatttg agcgaagatg tacaaaaaac tttcatcgaa gattggaatg aatacctaaa
     6901 agagaacaat ggcgatgtag ggacaatgga agaagagcag gtaattcatc cagtgattcg
     6961 caagcgttac gaagataaat tcaattattt tgccattcga tttttagatg agtttgctca
     7021 gttccctacg cttcgttttc aagtgcattt agggaattat cttcacgatt ctcgccctaa
     7081 agaaaatctg atttctgacc gaagaataaa agaaaaaatc accgttttcg gaaggctttc
     7141 ggaattggaa cataaaaaag cattgtttat aaaaaatacc gaaaccaatg aagatagaga
     7201 acactattgg gaaatttttc ctaaccccaa ttatgatttt cctaaggaaa atatttccgt
     7261 taatgataaa gattttccta tcgcgggtag tattttggac agagagaaac agcctgtcgc
     7321 agggaaaatt ggtataaaag tgaaattgct taaccaacaa tatgtttcag aagtagataa
     7381 agccgtaaaa gcccatcagc tcaagcaaag aaaagccagc aagccgtcta tccagaacat
     7441 tatagaagaa atagtgccaa tcaatgaaag caatccaaaa gaagctattg ttttcggtgg
     7501 gcaacccacg gcttatctga gtatgaatga cattcattcc attttgtatg agtttttcga
     7561 taaatgggag aaaaagaaag aaaagttaga aaagaaaggt gaaaaagaat taagaaagga
     7621 aattggaaaa gagttagaaa agaaaatcgt aggaaaaatt caagctcaaa tccagcaaat
     7681 catagataaa gataccaatg ccaagatatt aaaaccttat caagacggaa attctactgc
     7741 tattgataaa gagaaattga taaaagattt gaagcaagag caaaacattc tacaaaaatt
     7801 aaaagatgaa caaacggttc gtgaaaaaga atataatgat tttattgctt atcaagataa
     7861 aaatagggag ataaataaag tcagagatag aaatcataaa caatatttaa aagataacct
     7921 taaacgaaag taccctgaag ctcctgcacg aaaagaagtt ctttattatc gagaaaaagg
     7981 aaaagtagcg gtttggttgg cgaatgacat caaacgcttt atgcctaccg atttcaaaaa
     8041 cgaatggaaa ggggagcaac acagcttgtt acaaaaatcg ttggcgtatt atgagcaa
//
