LOCUS       sequence-325316-        7721 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible expression of BzCas13b nuclease in bacteria..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7721)
  AUTHORS   Wandera KG, Beisel CL
  TITLE     Rapidly Characterizing CRISPR-Cas13 Nucleases Using Cell-Free 
            Transcription-Translation Systems.
  JOURNAL   Methods Mol Biol. 2022;2404:135-153. doi: 
            10.1007/978-1-0716-1851-6_7.
  PUBMED    34694607
REFERENCE   2  (bases 1 to 7721)
  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: "Methods Mol
            Biol."; date: "2022"; volume: "2404:135-153. doi"; pages: " 
            10.1007/978-1-0716-1851-6_7"
COMMENT     Sequence Label: pBzCas13b
FEATURES             Location/Qualifiers
     source          1..7721
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             23..31
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     primer_bind     complement(3752..3769)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      3922..4008
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4100..4127
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        4146..4237
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4695..5150
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4782..4801)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4992..5013
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(5706..6365)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     6280..6299
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(6366..6468)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(6860..6877)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6994..7539)
                     /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(7074..7093)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7615..7634
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 gatgcattaa tacgactcac tataaggagg tctttatcat ggaaaacaaa acatctttgg
       61 ggaataatat ttattacaac ccttttaaac cacaagataa atcgtatttt gcgggttatt
      121 tcaatgcagc aatggaaaat acagatagtg tatttagaga attgggaaaa agattaaaag
      181 gtaaagagta cacttcggag aatttttttg atgctatctt taaggaaaat atttcattag
      241 tggaatacga aaggtatgtg aaattgcttt ctgattattt ccctatggct cgtcttttgg
      301 ataaaaaaga agttcctata aaagaacgaa aagaaaattt taaaaagaat tttaaaggca
      361 ttataaaagc cgtaagagat ttaagaaact tttatacgca taaggaacac ggagaggtag
      421 aaattacaga tgaaatattt ggcgtattgg atgagatgct gaaaagcacg gttttaacgg
      481 taaaaaagaa aaaagtaaaa accgataaaa caaaagaaat cctgaaaaaa agcatagaaa
      541 aacaattgga tattttatgc caaaagaaat tagaatatct aagagacaca gcaagaaaaa
      601 tagaggagaa gcgtagaaat caaagagaaa gaggagagaa agaattggta gctccattca
      661 aatacagtga taaaagggat gatttaatag cagcgattta taatgatgct tttgatgttt
      721 atatcgataa gaaaaaagat agcctaaaag aatccagcaa agcaaagtac aataccaaaa
      781 gtgatcctca gcaagaagaa ggcgatttga aaattccaat ttcgaaaaac ggagtagtct
      841 tcctgctttc tttgtttctc accaaacaag aaatacacgc attcaaatct aaaatagcag
      901 ggtttaaggc tacggttatc gatgaagcaa cggtatctga agcaacggtg agccatggaa
      961 aaaacagcat ttgttttatg gctacgcacg aaattttctc gcatttggct tacaaaaaat
     1021 tgaaacgaaa agtgagaacg gctgaaatca actatggtga ggcagaaaat gctgaacagc
     1081 tttccgttta tgccaaagaa actctaatga tgcagatgct tgatgaattg agcaaagtac
     1141 ccgatgtggt ctatcagaat ttgagcgaag atgtacaaaa aactttcatc gaagattgga
     1201 atgaatacct aaaagagaac aatggcgatg tagggacaat ggaagaagag caggtaattc
     1261 atccagtgat tcgcaagcgt tacgaagata aattcaatta ttttgccatt cgatttttag
     1321 atgagtttgc tcagttccct acgcttcgtt ttcaagtgca tttagggaat tatcttcacg
     1381 attctcgccc taaagaaaat ctgatttctg accgaagaat aaaagaaaaa atcaccgttt
     1441 tcggaaggct ttcggaattg gaacataaaa aagcattgtt tataaaaaat accgaaacca
     1501 atgaagatag agaacactat tgggaaattt ttcctaaccc caattatgat tttcctaagg
     1561 aaaatatttc cgttaatgat aaagattttc ctatcgcggg tagtattttg gacagagaga
     1621 aacagcctgt cgcagggaaa attggtataa aagtgaaatt gcttaaccaa caatatgttt
     1681 cagaagtaga taaagccgta aaagcccatc agctcaagca aagaaaagcc agcaagccgt
     1741 ctatccagaa cattatagaa gaaatagtgc caatcaatga aagcaatcca aaagaagcta
     1801 ttgttttcgg tgggcaaccc acggcttatc tgagtatgaa tgacattcat tccattttgt
     1861 atgagttttt cgataaatgg gagaaaaaga aagaaaagtt agaaaagaaa ggtgaaaaag
     1921 aattaagaaa ggaaattgga aaagagttag aaaagaaaat cgtaggaaaa attcaagctc
     1981 aaatccagca aatcatagat aaagatacca atgccaagat attaaaacct tatcaagacg
     2041 gaaattctac tgctattgat aaagagaaat tgataaaaga tttgaagcaa gagcaaaaca
     2101 ttctacaaaa attaaaagat gaacaaacgg ttcgtgaaaa agaatataat gattttattg
     2161 cttatcaaga taaaaatagg gagataaata aagtcagaga tagaaatcat aaacaatatt
     2221 taaaagataa ccttaaacga aagtaccctg aagctcctgc acgaaaagaa gttctttatt
     2281 atcgagaaaa aggaaaagta gcggtttggt tggcgaatga catcaaacgc tttatgccta
     2341 ccgatttcaa aaacgaatgg aaaggggagc aacacagctt gttacaaaaa tcgttggcgt
     2401 attatgagca atgtaaagag gagttgaaaa atcttttgcc tgaaaaagta tttcagcatt
     2461 taccttttaa gttgggagga tattttcaac aaaaatattt gtaccaattt tacacttgtt
     2521 atttagataa acggttggaa tacatcagtg ggttggtgca acaagccgaa aactttaaat
     2581 ccgaaaataa agtcttcaaa aaggtagaaa atgagtgttt taaatttttg aaaaaacaaa
     2641 attatactca caaagaatta gatgctcggg ttcagtctat attggggtat cctatctttt
     2701 tagagcgggg ctttatggat gaaaaaccta cgataatcaa aggaaaaact tttaaaggaa
     2761 atgaagctct ttttgccgat tggtttaggt attacaagga atatcagaat tttcagacat
     2821 tttatgatac tgaaaattat ccgcttgtag aattggaaaa aaaacaagca gaccgaaaac
     2881 gaaaaactaa aatctaccaa caaaagaaaa acgatgtgtt tacacttttg atggcaaaac
     2941 atattttcaa atcggtattc aagcaggata gcatcgatca gttttcgtta gaagatttgt
     3001 atcaatctcg ggaggaacgc ttaggaaatc aggagagagc cagacaaacg ggcgagcgaa
     3061 atactaacta tatttggaac aaaaccgttg atttaaagtt gtgtgacgga aaaattacgg
     3121 tagaaaatgt aaagctaaaa aatgtaggag atttcataaa atatgaatac gaccaaaggg
     3181 ttcaagcttt cttaaaatat gaggaaaata tagaatggca ggcgtttcta atcaaagaga
     3241 gtaaggaaga ggaaaattat ccgtatgtgg tggaaagaga aatagagcaa tatgaaaaag
     3301 tgagaaggga agaactgttg aaggaagtcc accttataga ggaatatatt ttggaaaagg
     3361 taaaagataa ggaaatatta aaaaaagggg ataatcaaaa tttcaaatac tacattctca
     3421 acggactttt aaagcaactt aaaaatgaag atgtggagag ctataaggta ttcaacttaa
     3481 atactgaacc tgaagatgtg aacattaacc aactaaaaca agaagcaacc gatttagaac
     3541 aaaaagcttt tgttttaacc tatattcgta ataaatttgc acacaaccaa ttacctaaaa
     3601 aagaattttg ggactattgt caagaaaaat acggcaaaat agaaaaagaa aaaacctatg
     3661 ccgagtattt tgctgaggta tttaaaaagg aaaaagaagc actgataaag taaaagcttg
     3721 gctgttttgg cggatgagag aagattttca gcctgataca gattaaatca gaacgcagaa
     3781 gcggtctgat aaaacagaat ttgcctggcg gcagtagcgc ggtggtccca cctgacccca
     3841 tgccgaactc agaagtgaaa cgccgtagcg ccgatggtag tgtggggtct ccccatgcga
     3901 gagtagggaa ctgccaggca tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt
     3961 cgttttatct gttgtttgtc ggtgaacgct ctcctgagta ggacaaatcc gccgggagcg
     4021 gatttgaacg ttgcgaagca acggcccgga gggtggcggg caggacgccc gccataaact
     4081 gccaggcatc aaattaagca gaaggccatc ctgacggatg gcctttttgc gtttctacaa
     4141 actcttttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     4201 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     4261 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     4321 ggtgaaagta aaagatgctg aagatcagtt gggtgcagca aactattaac tggcgaacta
     4381 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     4441 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     4501 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     4561 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     4621 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     4681 ctttagattg atttacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     4741 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     4801 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     4861 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt tgggtgatgg
     4921 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     4981 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcgggcta
     5041 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
     5101 ttaacaaaaa tttaacgcga attttaacaa aatattaacg tttacaattt aaaaggatct
     5161 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     5221 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     5281 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     5341 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     5401 atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     5461 ctacatacct cgctctgcta atcctgttac cagtcaggca tttgagaagc acacggtcac
     5521 actgcttccg gtagtcaata aaccggtaaa ccagcaatag acataagcgg ctatttaacg
     5581 accctgccct gaaccgacga ccgggtcgaa tttgctttcg aatttctgcc attcatccgc
     5641 ttattatcac ttattcaggc gtagcaacca ggcgtttaag ggcaccaata actgccttaa
     5701 aaaaattacg ccccgccctg ccactcatcg cagtactgtt gtaattcatt aagcattctg
     5761 ccgacatgga agccatcaca aacggcatga tgaacctgaa tcgccagcgg catcagcacc
     5821 ttgtcgcctt gcgtataata tttgcccatg gtgaaaacgg gggcgaagaa gttgtccata
     5881 ttggccacgt ttaaatcaaa actggtgaaa ctcacccagg gattggctga gacgaaaaac
     5941 atattctcaa taaacccttt agggaaatag gccaggtttt caccgtaaca cgccacatct
     6001 tgcgaatata tgtgtagaaa ctgccggaaa tcgtcgtggt attcactcca gagcgatgaa
     6061 aacgtttcag tttgctcatg gaaaacggtg taacaagggt gaacactatc ccatatcacc
     6121 agctcaccgt ctttcattgc catacggaat tccggatgag cattcatcag gcgggcaaga
     6181 atgtgaataa aggccggata aaacttgtgc ttatttttct ttacggtctt taaaaaggcc
     6241 gtaatatcca gctgaacggt ctggttatag gtacattgag caactgactg aaatgcctca
     6301 aaatgttctt tacgatgcca ttgggatata tcaacggtgg tatatccagt gatttttttc
     6361 tccattttag cttccttagc tcctgaaaat ctcgataact caaaaaatac gcccggtagt
     6421 gatcttattt cattatggtg aaagttggaa cctcttacgt gccgatcaac gtctcatttt
     6481 cgccaaaagt tggcccaggg cttcccggta tcaacaggga caccaggatt tatttattct
     6541 gcgaagtgat cttccgtcac aggtatttat tcggcgcaaa gtgcgtcggg tgatgctgcc
     6601 aacttactga tttagtgtat gatggtgttt ttgaggtgct ccagtggctt ctgtttctat
     6661 cagctgtccc tcctgttcag ctactgacgg ggtggtgcgt aacggcaaaa gcaccgccgg
     6721 acatcagcgc tagcggagtg tatactggct tactatgttg gcactgatga gggtgtcagt
     6781 gaagtgcttc atgtggcagg agaaaaaagg ctgcaccggt gcgtcagcag aatatgtgat
     6841 acaggatata ttccgcttcc tcgctcactg actcgctacg ctcggtcgtt cgactgcggc
     6901 gagcggaaat ggcttacgaa cggggcggag atttcctgga agatgccagg aagatactta
     6961 acagggaagt gagagggccg cggcaaagcc gtttttccat aggctccgcc cccctgacaa
     7021 gcatcacgaa atctgacgct caaatcagtg gtggcgaaac ccgacaggac tataaagata
     7081 ccaggcgttt ccccctggcg gctccctcgt gcgctctcct gttcctgcct ttcggtttac
     7141 cggtgtcatt ccgctgttat ggccgcgttt gtctcattcc acgcctgaca ctcagttccg
     7201 ggtaggcagt tcgctccaag ctggactgta tgcacgaacc ccccgttcag tccgaccgct
     7261 gcgccttatc cggtaactat cgtcttgagt ccaacccgga aagacatgca aaagcaccac
     7321 tggcagcagc cactggtaat tgatttagag gagttagtct tgaagtcatg cgccggttaa
     7381 ggctaaactg aaaggacaag ttttggtgac tgcgctcctc caagccagtt acctcggttc
     7441 aaagagttgg tagctcagag aaccttcgaa aaaccgccct gcaaggcggt tttttcgttt
     7501 tcagagcaag agattacgcg cagaccaaaa cgatctcaag aagatcatct tattaatcag
     7561 ataaaatatt tctaggctca tgagcccgaa gtggcgagcc cgatcttccc catcggtgat
     7621 gtcggcgata taggcgccag caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg
     7681 tccggcgtag aggatctgct catgtttgac agcttatcat c
//
