LOCUS       sequence-377030-        4360 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Constitutive expression of single-spacer CRISPR array with spacer #1
            targeting deGFP mRNA 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-1
FEATURES             Location/Qualifiers
     source          1..4360
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(293..952)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     867..886
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(953..1055)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(1447..1464)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1581..2126)
                     /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(1661..1680)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        join(2238..2243,2244..2260,2261..2266)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     promoter        2389..2423
                     /label=J23119 promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119)"
     CDS             complement(2435..2446)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     2465..2486
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             complement(2501..2512)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2781..2800)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 tttatcaggc tctgggaggc agaataaatg atcatatcgt caattattac ctccacgggg
       61 agagcctgag caaactggcc tcaggcattt gagaagcaca cggtcacact gcttccggta
      121 gtcaataaac cggtaaacca gcaatagaca taagcggcta tttaacgacc ctgccctgaa
      181 ccgacgaccg ggtcgaattt gctttcgaat ttctgccatt catccgctta ttatcactta
      241 ttcaggcgta gcaaccaggc gtttaagggc accaataact gccttaaaaa aattacgccc
      301 cgccctgcca ctcatcgcag tactgttgta attcattaag cattctgccg acatggaagc
      361 catcacaaac ggcatgatga acctgaatcg ccagcggcat cagcaccttg tcgccttgcg
      421 tataatattt gcccatggtg aaaacggggg cgaagaagtt gtccatattg gccacgttta
      481 aatcaaaact ggtgaaactc acccagggat tggctgagac gaaaaacata ttctcaataa
      541 accctttagg gaaataggcc aggttttcac cgtaacacgc cacatcttgc gaatatatgt
      601 gtagaaactg ccggaaatcg tcgtggtatt cactccagag cgatgaaaac gtttcagttt
      661 gctcatggaa aacggtgtaa caagggtgaa cactatccca tatcaccagc tcaccgtctt
      721 tcattgccat acggaattcc ggatgagcat tcatcaggcg ggcaagaatg tgaataaagg
      781 ccggataaaa cttgtgctta tttttcttta cggtctttaa aaaggccgta atatccagct
      841 gaacggtctg gttataggta cattgagcaa ctgactgaaa tgcctcaaaa tgttctttac
      901 gatgccattg ggatatatca acggtggtat atccagtgat ttttttctcc attttagctt
      961 ccttagctcc tgaaaatctc gataactcaa aaaatacgcc cggtagtgat cttatttcat
     1021 tatggtgaaa gttggaacct cttacgtgcc gatcaacgtc tcattttcgc caaaagttgg
     1081 cccagggctt cccggtatca acagggacac caggatttat ttattctgcg aagtgatctt
     1141 ccgtcacagg tatttattcg gcgcaaagtg cgtcgggtga tgctgccaac ttactgattt
     1201 agtgtatgat ggtgtttttg aggtgctcca gtggcttctg tttctatcag ctgtccctcc
     1261 tgttcagcta ctgacggggt ggtgcgtaac ggcaaaagca ccgccggaca tcagcgctag
     1321 cggagtgtat actggcttac tatgttggca ctgatgaggg tgtcagtgaa gtgcttcatg
     1381 tggcaggaga aaaaaggctg caccggtgcg tcagcagaat atgtgataca ggatatattc
     1441 cgcttcctcg ctcactgact cgctacgctc ggtcgttcga ctgcggcgag cggaaatggc
     1501 ttacgaacgg ggcggagatt tcctggaaga tgccaggaag atacttaaca gggaagtgag
     1561 agggccgcgg caaagccgtt tttccatagg ctccgccccc ctgacaagca tcacgaaatc
     1621 tgacgctcaa atcagtggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     1681 cctggcggct ccctcgtgcg ctctcctgtt cctgcctttc ggtttaccgg tgtcattccg
     1741 ctgttatggc cgcgtttgtc tcattccacg cctgacactc agttccgggt aggcagttcg
     1801 ctccaagctg gactgtatgc acgaaccccc cgttcagtcc gaccgctgcg ccttatccgg
     1861 taactatcgt cttgagtcca acccggaaag acatgcaaaa gcaccactgg cagcagccac
     1921 tggtaattga tttagaggag ttagtcttga agtcatgcgc cggttaaggc taaactgaaa
     1981 ggacaagttt tggtgactgc gctcctccaa gccagttacc tcggttcaaa gagttggtag
     2041 ctcagagaac cttcgaaaaa ccgccctgca aggcggtttt ttcgttttca gagcaagaga
     2101 ttacgcgcag accaaaacga tctcaagaag atcatcttat taatcagata aaatatttct
     2161 agatttcagt gcaatttatc tcttcaaatg tagcacctga agtcagcccc atacgatata
     2221 agttgtaatt ctcatgtttg acagcttatc atcgataagc tttaatgcgg tagtttatca
     2281 cagttaaatt gctaacgcag tcaggcaccg tgtatgaaat ctaacaatgc gctcatcgtc
     2341 atcctcggca ccgtcaccct ggatgctgta ggcataggct tggtttaatt gacagctagc
     2401 tcagtcctag gtataatgct agcgctgagt tggatctacc ctctattcga agggtacaca
     2461 caaccgtcgc cgtccagctc gaccaggatg ggaagttgga tctaccctct attcgaaggg
     2521 tacacacaac atcgtccatt ccgacagcat cgccagtcac tatggcgtgc tgctagcgct
     2581 atatgcgttg atgcaatttc tatgcgcacc cgttctcgga gcactgtccg accgctttgg
     2641 ccgccgccca gtcctgctcg cttcgctact tggagccact atcgactacg cgatcatggc
     2701 gaccacaccc gtcctgtgga tcctctacgc cggacgcatc gtggccggca tcaccggcgc
     2761 cacaggtgcg gttgctggcg cctatatcgc cgacatcacc gatggggaag atcgggctcg
     2821 ccacttcggg ctcatgagcg cttgtttcgg cgtgggtatg gtggcaggcc ccgtggccgg
     2881 gggactgttg ggcgccatct ccttgcatgc accattcctt gcggcggcgg tgctcaacgg
     2941 cctcaaccta ctactgggct gcttcctaat gcaggagtcg cataagggag agcgtcgacc
     3001 gatgcccttg agagccttca acccagtcag ctccttccgg tgggcgcggg gcatgactat
     3061 cgtcgccgca cttatgactg tcttctttat catgcaactc gtaggacagg tgccggcagc
     3121 gctctgggtc attttcggcg aggaccgctt tcgctggagc gcgacgatga tcggcctgtc
     3181 gcttgcggta ttcggaatct tgcacgccct cgctcaagcc ttcgtcactg gtcccgccac
     3241 caaacgtttc ggcgagaagc aggccattat cgccggcatg gcggccgacg cgctgggcta
     3301 cgtcttgctg gcgttcgcga cgcgaggctg gatggccttc cccattatga ttcttctcgc
     3361 ttccggcggc atcgggatgc ccgcgttgca ggccatgctg tccaggcagg tagatgacga
     3421 ccatcaggga cagcttcaag gatcgctcgc ggctcttacc agcctaactt cgatcattgg
     3481 accgctgatc gtcacggcga tttatgccgc ctcggcgagc acatggaacg ggttggcatg
     3541 gattgtaggc gccgccctat accttgtctg cctccccgcg ttgcgtcgcg gtgcatggag
     3601 ccgggccacc tcgacctgaa tggaagccgg cggcacctcg ctaacggatt caccactcca
     3661 agaattggag ccaatcaatt cttgcggaga actgtgaatg cgcaaaccaa cccttggcag
     3721 aacatatcca tcgcgtccgc catctccagc agccgcacgc ggcgcatctc gggcagcgtt
     3781 gggtcctggc cacgggtgcg catgatcgtg ctcctgtcgt tgaggacccg gctaggctgg
     3841 cggggttgcc ttactggtta gcagaatgaa tcaccgatac gcgagcgaac gtgaagcgac
     3901 tgctgctgca aaacgtctgc gacctgagca acaacatgaa tggtcttcgg tttccgtgtt
     3961 tcgtaaagtc tggaaacgcg gaagtcccct acgtgctgct gaagttgccc gcaacagaga
     4021 gtggaaccaa ccggtgatac cacgatacta tgactgagag tcaacgccat gagcggcctc
     4081 atttcttatt ctgagttaca acagtccgca ccgctgccgg tagctccttc cggtgggcgc
     4141 ggggcatgac tatcgtcgcc gcacttatga ctgtcttctt tatcatgcaa ctcgtaggac
     4201 aggtgccggc agcgcccaac agtcccccgg ccacggggcc tgccaccata cccacgccga
     4261 aacaagcgcc ctgcaccatt atgttccgga tctgcatcgc aggatgctgc tggctaccct
     4321 gtggaacacc tacatctgta ttaacgaagc gctaaccgtt
//
