LOCUS       sequence-377028-        4360 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Constitutive expression of single-spacer CRISPR array with 
            non-targeting spacer for PgiCas13b 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: pPgiCas13b-gRNA-NT
FEATURES             Location/Qualifiers
     source          1..4360
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(289..948)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     863..882
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(949..1051)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(1443..1460)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1577..2122)
                     /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(1657..1676)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        join(2234..2239,2240..2256,2257..2262)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     promoter        2385..2419
                     /label=J23119 promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119)"
     CDS             complement(2431..2442)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             complement(2497..2508)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2777..2796)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 tcaggctctg ggaggcagaa taaatgatca tatcgtcaat tattacctcc acggggagag
       61 cctgagcaaa ctggcctcag gcatttgaga agcacacggt cacactgctt ccggtagtca
      121 ataaaccggt aaaccagcaa tagacataag cggctattta acgaccctgc cctgaaccga
      181 cgaccgggtc gaatttgctt tcgaatttct gccattcatc cgcttattat cacttattca
      241 ggcgtagcaa ccaggcgttt aagggcacca ataactgcct taaaaaaatt acgccccgcc
      301 ctgccactca tcgcagtact gttgtaattc attaagcatt ctgccgacat ggaagccatc
      361 acaaacggca tgatgaacct gaatcgccag cggcatcagc accttgtcgc cttgcgtata
      421 atatttgccc atggtgaaaa cgggggcgaa gaagttgtcc atattggcca cgtttaaatc
      481 aaaactggtg aaactcaccc agggattggc tgagacgaaa aacatattct caataaaccc
      541 tttagggaaa taggccaggt tttcaccgta acacgccaca tcttgcgaat atatgtgtag
      601 aaactgccgg aaatcgtcgt ggtattcact ccagagcgat gaaaacgttt cagtttgctc
      661 atggaaaacg gtgtaacaag ggtgaacact atcccatatc accagctcac cgtctttcat
      721 tgccatacgg aattccggat gagcattcat caggcgggca agaatgtgaa taaaggccgg
      781 ataaaacttg tgcttatttt tctttacggt ctttaaaaag gccgtaatat ccagctgaac
      841 ggtctggtta taggtacatt gagcaactga ctgaaatgcc tcaaaatgtt ctttacgatg
      901 ccattgggat atatcaacgg tggtatatcc agtgattttt ttctccattt tagcttcctt
      961 agctcctgaa aatctcgata actcaaaaaa tacgcccggt agtgatctta tttcattatg
     1021 gtgaaagttg gaacctctta cgtgccgatc aacgtctcat tttcgccaaa agttggccca
     1081 gggcttcccg gtatcaacag ggacaccagg atttatttat tctgcgaagt gatcttccgt
     1141 cacaggtatt tattcggcgc aaagtgcgtc gggtgatgct gccaacttac tgatttagtg
     1201 tatgatggtg tttttgaggt gctccagtgg cttctgtttc tatcagctgt ccctcctgtt
     1261 cagctactga cggggtggtg cgtaacggca aaagcaccgc cggacatcag cgctagcgga
     1321 gtgtatactg gcttactatg ttggcactga tgagggtgtc agtgaagtgc ttcatgtggc
     1381 aggagaaaaa aggctgcacc ggtgcgtcag cagaatatgt gatacaggat atattccgct
     1441 tcctcgctca ctgactcgct acgctcggtc gttcgactgc ggcgagcgga aatggcttac
     1501 gaacggggcg gagatttcct ggaagatgcc aggaagatac ttaacaggga agtgagaggg
     1561 ccgcggcaaa gccgtttttc cataggctcc gcccccctga caagcatcac gaaatctgac
     1621 gctcaaatca gtggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     1681 gcggctccct cgtgcgctct cctgttcctg cctttcggtt taccggtgtc attccgctgt
     1741 tatggccgcg tttgtctcat tccacgcctg acactcagtt ccgggtaggc agttcgctcc
     1801 aagctggact gtatgcacga accccccgtt cagtccgacc gctgcgcctt atccggtaac
     1861 tatcgtcttg agtccaaccc ggaaagacat gcaaaagcac cactggcagc agccactggt
     1921 aattgattta gaggagttag tcttgaagtc atgcgccggt taaggctaaa ctgaaaggac
     1981 aagttttggt gactgcgctc ctccaagcca gttacctcgg ttcaaagagt tggtagctca
     2041 gagaaccttc gaaaaaccgc cctgcaaggc ggttttttcg ttttcagagc aagagattac
     2101 gcgcagacca aaacgatctc aagaagatca tcttattaat cagataaaat atttctagat
     2161 ttcagtgcaa tttatctctt caaatgtagc acctgaagtc agccccatac gatataagtt
     2221 gtaattctca tgtttgacag cttatcatcg ataagcttta atgcggtagt ttatcacagt
     2281 taaattgcta acgcagtcag gcaccgtgta tgaaatctaa caatgcgctc atcgtcatcc
     2341 tcggcaccgt caccctggat gctgtaggca taggcttggt ttaattgaca gctagctcag
     2401 tcctaggtat aatgctagcg ctgagttgga tctaccctct attcgaaggg tacacacaac
     2461 ctcgtatgtt gtgtggaatt gtgagcggat gttggatcta ccctctattc gaagggtaca
     2521 cacaacatcg tccattccga cagcatcgcc agtcactatg gcgtgctgct agcgctatat
     2581 gcgttgatgc aatttctatg cgcacccgtt ctcggagcac tgtccgaccg ctttggccgc
     2641 cgcccagtcc tgctcgcttc gctacttgga gccactatcg actacgcgat catggcgacc
     2701 acacccgtcc tgtggatcct ctacgccgga cgcatcgtgg ccggcatcac cggcgccaca
     2761 ggtgcggttg ctggcgccta tatcgccgac atcaccgatg gggaagatcg ggctcgccac
     2821 ttcgggctca tgagcgcttg tttcggcgtg ggtatggtgg caggccccgt ggccggggga
     2881 ctgttgggcg ccatctcctt gcatgcacca ttccttgcgg cggcggtgct caacggcctc
     2941 aacctactac tgggctgctt cctaatgcag gagtcgcata agggagagcg tcgaccgatg
     3001 cccttgagag ccttcaaccc agtcagctcc ttccggtggg cgcggggcat gactatcgtc
     3061 gccgcactta tgactgtctt ctttatcatg caactcgtag gacaggtgcc ggcagcgctc
     3121 tgggtcattt tcggcgagga ccgctttcgc tggagcgcga cgatgatcgg cctgtcgctt
     3181 gcggtattcg gaatcttgca cgccctcgct caagccttcg tcactggtcc cgccaccaaa
     3241 cgtttcggcg agaagcaggc cattatcgcc ggcatggcgg ccgacgcgct gggctacgtc
     3301 ttgctggcgt tcgcgacgcg aggctggatg gccttcccca ttatgattct tctcgcttcc
     3361 ggcggcatcg ggatgcccgc gttgcaggcc atgctgtcca ggcaggtaga tgacgaccat
     3421 cagggacagc ttcaaggatc gctcgcggct cttaccagcc taacttcgat cattggaccg
     3481 ctgatcgtca cggcgattta tgccgcctcg gcgagcacat ggaacgggtt ggcatggatt
     3541 gtaggcgccg ccctatacct tgtctgcctc cccgcgttgc gtcgcggtgc atggagccgg
     3601 gccacctcga cctgaatgga agccggcggc acctcgctaa cggattcacc actccaagaa
     3661 ttggagccaa tcaattcttg cggagaactg tgaatgcgca aaccaaccct tggcagaaca
     3721 tatccatcgc gtccgccatc tccagcagcc gcacgcggcg catctcgggc agcgttgggt
     3781 cctggccacg ggtgcgcatg atcgtgctcc tgtcgttgag gacccggcta ggctggcggg
     3841 gttgccttac tggttagcag aatgaatcac cgatacgcga gcgaacgtga agcgactgct
     3901 gctgcaaaac gtctgcgacc tgagcaacaa catgaatggt cttcggtttc cgtgtttcgt
     3961 aaagtctgga aacgcggaag tcccctacgt gctgctgaag ttgcccgcaa cagagagtgg
     4021 aaccaaccgg tgataccacg atactatgac tgagagtcaa cgccatgagc ggcctcattt
     4081 cttattctga gttacaacag tccgcaccgc tgccggtagc tccttccggt gggcgcgggg
     4141 catgactatc gtcgccgcac ttatgactgt cttctttatc atgcaactcg taggacaggt
     4201 gccggcagcg cccaacagtc ccccggccac ggggcctgcc accataccca cgccgaaaca
     4261 agcgccctgc accattatgt tccggatctg catcgcagga tgctgctggc taccctgtgg
     4321 aacacctaca tctgtattaa cgaagcgcta accgttttta
//
