LOCUS       sequence-325312-        4360 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Constitutive expression of single-spacer CRISPR array with 
            non-targeting spacer for BzCas13b in bacteria..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4360)
  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 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: "Methods Mol
            Biol."; date: "2022"; volume: "2404:135-153. doi"; pages: " 
            10.1007/978-1-0716-1851-6_7"
COMMENT     Sequence Label: pBzCas13b-gRNA-NT
FEATURES             Location/Qualifiers
     source          1..4360
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     131..150
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(217..319)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(711..728)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(845..1390)
                     /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(925..944)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        join(1502..1507,1508..1524,1525..1530)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     promoter        1653..1687
                     /label=J23119 promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119)"
     primer_bind     complement(2045..2064)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(join(3917..4360,1..216))
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
ORIGIN
        1 ttccggatga gcattcatca ggcgggcaag aatgtgaata aaggccggat aaaacttgtg
       61 cttatttttc tttacggtct ttaaaaaggc cgtaatatcc agctgaacgg tctggttata
      121 ggtacattga gcaactgact gaaatgcctc aaaatgttct ttacgatgcc attgggatat
      181 atcaacggtg gtatatccag tgattttttt ctccatttta gcttccttag ctcctgaaaa
      241 tctcgataac tcaaaaaata cgcccggtag tgatcttatt tcattatggt gaaagttgga
      301 acctcttacg tgccgatcaa cgtctcattt tcgccaaaag ttggcccagg gcttcccggt
      361 atcaacaggg acaccaggat ttatttattc tgcgaagtga tcttccgtca caggtattta
      421 ttcggcgcaa agtgcgtcgg gtgatgctgc caacttactg atttagtgta tgatggtgtt
      481 tttgaggtgc tccagtggct tctgtttcta tcagctgtcc ctcctgttca gctactgacg
      541 gggtggtgcg taacggcaaa agcaccgccg gacatcagcg ctagcggagt gtatactggc
      601 ttactatgtt ggcactgatg agggtgtcag tgaagtgctt catgtggcag gagaaaaaag
      661 gctgcaccgg tgcgtcagca gaatatgtga tacaggatat attccgcttc ctcgctcact
      721 gactcgctac gctcggtcgt tcgactgcgg cgagcggaaa tggcttacga acggggcgga
      781 gatttcctgg aagatgccag gaagatactt aacagggaag tgagagggcc gcggcaaagc
      841 cgtttttcca taggctccgc ccccctgaca agcatcacga aatctgacgc tcaaatcagt
      901 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctggc ggctccctcg
      961 tgcgctctcc tgttcctgcc tttcggttta ccggtgtcat tccgctgtta tggccgcgtt
     1021 tgtctcattc cacgcctgac actcagttcc gggtaggcag ttcgctccaa gctggactgt
     1081 atgcacgaac cccccgttca gtccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     1141 tccaacccgg aaagacatgc aaaagcacca ctggcagcag ccactggtaa ttgatttaga
     1201 ggagttagtc ttgaagtcat gcgccggtta aggctaaact gaaaggacaa gttttggtga
     1261 ctgcgctcct ccaagccagt tacctcggtt caaagagttg gtagctcaga gaaccttcga
     1321 aaaaccgccc tgcaaggcgg ttttttcgtt ttcagagcaa gagattacgc gcagaccaaa
     1381 acgatctcaa gaagatcatc ttattaatca gataaaatat ttctagattt cagtgcaatt
     1441 tatctcttca aatgtagcac ctgaagtcag ccccatacga tataagttgt aattctcatg
     1501 tttgacagct tatcatcgat aagctttaat gcggtagttt atcacagtta aattgctaac
     1561 gcagtcaggc accgtgtatg aaatctaaca atgcgctcat cgtcatcctc ggcaccgtca
     1621 ccctggatgc tgtaggcata ggcttggttt aattgacagc tagctcagtc ctaggtataa
     1681 tgctagcgct gagttggaac tgctctcatt ttggagggta atcacaacct cgtatgttgt
     1741 gtggaattgt gagcggatgt tggaactgct ctcattttgg agggtaatca caacatcgtc
     1801 cattccgaca gcatcgccag tcactatggc gtgctgctag cgctatatgc gttgatgcaa
     1861 tttctatgcg cacccgttct cggagcactg tccgaccgct ttggccgccg cccagtcctg
     1921 ctcgcttcgc tacttggagc cactatcgac tacgcgatca tggcgaccac acccgtcctg
     1981 tggatcctct acgccggacg catcgtggcc ggcatcaccg gcgccacagg tgcggttgct
     2041 ggcgcctata tcgccgacat caccgatggg gaagatcggg ctcgccactt cgggctcatg
     2101 agcgcttgtt tcggcgtggg tatggtggca ggccccgtgg ccgggggact gttgggcgcc
     2161 atctccttgc atgcaccatt ccttgcggcg gcggtgctca acggcctcaa cctactactg
     2221 ggctgcttcc taatgcagga gtcgcataag ggagagcgtc gaccgatgcc cttgagagcc
     2281 ttcaacccag tcagctcctt ccggtgggcg cggggcatga ctatcgtcgc cgcacttatg
     2341 actgtcttct ttatcatgca actcgtagga caggtgccgg cagcgctctg ggtcattttc
     2401 ggcgaggacc gctttcgctg gagcgcgacg atgatcggcc tgtcgcttgc ggtattcgga
     2461 atcttgcacg ccctcgctca agccttcgtc actggtcccg ccaccaaacg tttcggcgag
     2521 aagcaggcca ttatcgccgg catggcggcc gacgcgctgg gctacgtctt gctggcgttc
     2581 gcgacgcgag gctggatggc cttccccatt atgattcttc tcgcttccgg cggcatcggg
     2641 atgcccgcgt tgcaggccat gctgtccagg caggtagatg acgaccatca gggacagctt
     2701 caaggatcgc tcgcggctct taccagccta acttcgatca ttggaccgct gatcgtcacg
     2761 gcgatttatg ccgcctcggc gagcacatgg aacgggttgg catggattgt aggcgccgcc
     2821 ctataccttg tctgcctccc cgcgttgcgt cgcggtgcat ggagccgggc cacctcgacc
     2881 tgaatggaag ccggcggcac ctcgctaacg gattcaccac tccaagaatt ggagccaatc
     2941 aattcttgcg gagaactgtg aatgcgcaaa ccaacccttg gcagaacata tccatcgcgt
     3001 ccgccatctc cagcagccgc acgcggcgca tctcgggcag cgttgggtcc tggccacggg
     3061 tgcgcatgat cgtgctcctg tcgttgagga cccggctagg ctggcggggt tgccttactg
     3121 gttagcagaa tgaatcaccg atacgcgagc gaacgtgaag cgactgctgc tgcaaaacgt
     3181 ctgcgacctg agcaacaaca tgaatggtct tcggtttccg tgtttcgtaa agtctggaaa
     3241 cgcggaagtc ccctacgtgc tgctgaagtt gcccgcaaca gagagtggaa ccaaccggtg
     3301 ataccacgat actatgactg agagtcaacg ccatgagcgg cctcatttct tattctgagt
     3361 tacaacagtc cgcaccgctg ccggtagctc cttccggtgg gcgcggggca tgactatcgt
     3421 cgccgcactt atgactgtct tctttatcat gcaactcgta ggacaggtgc cggcagcgcc
     3481 caacagtccc ccggccacgg ggcctgccac catacccacg ccgaaacaag cgccctgcac
     3541 cattatgttc cggatctgca tcgcaggatg ctgctggcta ccctgtggaa cacctacatc
     3601 tgtattaacg aagcgctaac cgtttttatc aggctctggg aggcagaata aatgatcata
     3661 tcgtcaatta ttacctccac ggggagagcc tgagcaaact ggcctcaggc atttgagaag
     3721 cacacggtca cactgcttcc ggtagtcaat aaaccggtaa accagcaata gacataagcg
     3781 gctatttaac gaccctgccc tgaaccgacg accgggtcga atttgctttc gaatttctgc
     3841 cattcatccg cttattatca cttattcagg cgtagcaacc aggcgtttaa gggcaccaat
     3901 aactgcctta aaaaaattac gccccgccct gccactcatc gcagtactgt tgtaattcat
     3961 taagcattct gccgacatgg aagccatcac aaacggcatg atgaacctga atcgccagcg
     4021 gcatcagcac cttgtcgcct tgcgtataat atttgcccat ggtgaaaacg ggggcgaaga
     4081 agttgtccat attggccacg tttaaatcaa aactggtgaa actcacccag ggattggctg
     4141 agacgaaaaa catattctca ataaaccctt tagggaaata ggccaggttt tcaccgtaac
     4201 acgccacatc ttgcgaatat atgtgtagaa actgccggaa atcgtcgtgg tattcactcc
     4261 agagcgatga aaacgtttca gtttgctcat ggaaaacggt gtaacaaggg tgaacactat
     4321 cccatatcac cagctcaccg tctttcattg ccatacggaa
//
