LOCUS       sequence-373995-        7406 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible expression of PgiCas13b nuclease in bacteria..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7406)
  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 7406)
  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
FEATURES             Location/Qualifiers
     source          1..7406
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(117..134)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      287..373
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      465..492
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        511..602
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      1060..1515
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1147..1166)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1357..1378
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2071..2730)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2645..2664
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(2731..2833)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(3225..3242)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3359..3904)
                     /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(3439..3458)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3980..3999
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     RBS             4109..4117
                     /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)"
ORIGIN
        1 ggcttaaaca tcgcacatcg tttatccgaa gaagtcaagc aggcaaaaga gatggttgaa
       61 cgtattattc aagcttaaaa gcttggctgt tttggcggat gagagaagat tttcagcctg
      121 atacagatta aatcagaacg cagaagcggt ctgataaaac agaatttgcc tggcggcagt
      181 agcgcggtgg tcccacctga ccccatgccg aactcagaag tgaaacgccg tagcgccgat
      241 ggtagtgtgg ggtctcccca tgcgagagta gggaactgcc aggcatcaaa taaaacgaaa
      301 ggctcagtcg aaagactggg cctttcgttt tatctgttgt ttgtcggtga acgctctcct
      361 gagtaggaca aatccgccgg gagcggattt gaacgttgcg aagcaacggc ccggagggtg
      421 gcgggcagga cgcccgccat aaactgccag gcatcaaatt aagcagaagg ccatcctgac
      481 ggatggcctt tttgcgtttc tacaaactct tttgtttatt tttctaaata cattcaaata
      541 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
      601 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
      661 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
      721 cagcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
      781 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
      841 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
      901 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
      961 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     1021 gtcagaccaa gtttactcat atatacttta gattgattta cgcgccctgt agcggcgcat
     1081 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     1141 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     1201 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     1261 ccaaaaaact tgatttgggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     1321 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     1381 caacactcaa ccctatctcg ggctattctt ttgatttata agggattttg ccgatttcgg
     1441 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     1501 taacgtttac aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     1561 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     1621 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     1681 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     1741 ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac
     1801 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtc
     1861 aggcatttga gaagcacacg gtcacactgc ttccggtagt caataaaccg gtaaaccagc
     1921 aatagacata agcggctatt taacgaccct gccctgaacc gacgaccggg tcgaatttgc
     1981 tttcgaattt ctgccattca tccgcttatt atcacttatt caggcgtagc aaccaggcgt
     2041 ttaagggcac caataactgc cttaaaaaaa ttacgccccg ccctgccact catcgcagta
     2101 ctgttgtaat tcattaagca ttctgccgac atggaagcca tcacaaacgg catgatgaac
     2161 ctgaatcgcc agcggcatca gcaccttgtc gccttgcgta taatatttgc ccatggtgaa
     2221 aacgggggcg aagaagttgt ccatattggc cacgtttaaa tcaaaactgg tgaaactcac
     2281 ccagggattg gctgagacga aaaacatatt ctcaataaac cctttaggga aataggccag
     2341 gttttcaccg taacacgcca catcttgcga atatatgtgt agaaactgcc ggaaatcgtc
     2401 gtggtattca ctccagagcg atgaaaacgt ttcagtttgc tcatggaaaa cggtgtaaca
     2461 agggtgaaca ctatcccata tcaccagctc accgtctttc attgccatac ggaattccgg
     2521 atgagcattc atcaggcggg caagaatgtg aataaaggcc ggataaaact tgtgcttatt
     2581 tttctttacg gtctttaaaa aggccgtaat atccagctga acggtctggt tataggtaca
     2641 ttgagcaact gactgaaatg cctcaaaatg ttctttacga tgccattggg atatatcaac
     2701 ggtggtatat ccagtgattt ttttctccat tttagcttcc ttagctcctg aaaatctcga
     2761 taactcaaaa aatacgcccg gtagtgatct tatttcatta tggtgaaagt tggaacctct
     2821 tacgtgccga tcaacgtctc attttcgcca aaagttggcc cagggcttcc cggtatcaac
     2881 agggacacca ggatttattt attctgcgaa gtgatcttcc gtcacaggta tttattcggc
     2941 gcaaagtgcg tcgggtgatg ctgccaactt actgatttag tgtatgatgg tgtttttgag
     3001 gtgctccagt ggcttctgtt tctatcagct gtccctcctg ttcagctact gacggggtgg
     3061 tgcgtaacgg caaaagcacc gccggacatc agcgctagcg gagtgtatac tggcttacta
     3121 tgttggcact gatgagggtg tcagtgaagt gcttcatgtg gcaggagaaa aaaggctgca
     3181 ccggtgcgtc agcagaatat gtgatacagg atatattccg cttcctcgct cactgactcg
     3241 ctacgctcgg tcgttcgact gcggcgagcg gaaatggctt acgaacgggg cggagatttc
     3301 ctggaagatg ccaggaagat acttaacagg gaagtgagag ggccgcggca aagccgtttt
     3361 tccataggct ccgcccccct gacaagcatc acgaaatctg acgctcaaat cagtggtggc
     3421 gaaacccgac aggactataa agataccagg cgtttccccc tggcggctcc ctcgtgcgct
     3481 ctcctgttcc tgcctttcgg tttaccggtg tcattccgct gttatggccg cgtttgtctc
     3541 attccacgcc tgacactcag ttccgggtag gcagttcgct ccaagctgga ctgtatgcac
     3601 gaaccccccg ttcagtccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     3661 ccggaaagac atgcaaaagc accactggca gcagccactg gtaattgatt tagaggagtt
     3721 agtcttgaag tcatgcgccg gttaaggcta aactgaaagg acaagttttg gtgactgcgc
     3781 tcctccaagc cagttacctc ggttcaaaga gttggtagct cagagaacct tcgaaaaacc
     3841 gccctgcaag gcggtttttt cgttttcaga gcaagagatt acgcgcagac caaaacgatc
     3901 tcaagaagat catcttatta atcagataaa atatttctag gctcatgagc ccgaagtggc
     3961 gagcccgatc ttccccatcg gtgatgtcgg cgatataggc gccagcaacc gcacctgtgg
     4021 cgccggtgat gccggccacg atgcgtccgg cgtagaggat ctgctcatgt ttgacagctt
     4081 atcatcgatg cattaatacg actcactata aggaggtctt tatcatgacg gagcagaacg
     4141 agaaacccta taacggcaca tattacacat tagaagacaa acatttctgg gcagcatttt
     4201 tgaaccttgc tcgtcataac gcctacatca ctttggccca cattgatcgc cagcttgcct
     4261 actccaaggc ggatatcacg aacgatgagg acatcctgtt cttcaaaggc caatggaaga
     4321 atttggacaa tgatcttgag cgtaaagcgc gcctgcgtag cttgatcttg aagcactttt
     4381 cctttcttga aggggctgcc tacgggaaga agttattcga atctcagtca agcgggaaca
     4441 agtcatctaa aaaaaaggaa ctgtccaaaa aggaaaaaga agagttgcaa gccaatgcgt
     4501 tgagcctgga caaccttaaa agtattttat ttgactttct gcagaagtta aaggattttc
     4561 gtaactacta tagccactat cgtcacccgg aaagctcgga gcttccctta tttgatggca
     4621 atatgttgca acgcctgtat aacgtatttg atgtgtcggt ccagcgcgtt aagcgtgacc
     4681 acgagcacaa cgacaaggtt gatcctcatc gccattttaa tcacttagtt cgcaaaggca
     4741 aaaaggacaa atacggtaat aatgataatc cattcttcaa acatcacttc gttgaccgcg
     4801 aggggacagt tacagaggca ggtcttctgt tctttgtctc attattcctt gagaaacgtg
     4861 acgcaatttg gatgcaaaag aaaatccgtg ggtttaaggg aggtacggaa gcctaccagc
     4921 aaatgactaa cgaggttttc tgccgttcgc gcattagcct tcctaaactt aaattagaat
     4981 ccttgcgtac ggatgactgg atgctgctgg acatgcttaa cgaattagta cgttgtccta
     5041 agagtctgta tgaccgtctt cgtgaagaag accgcgctcg ctttcgtgta cctgtagaca
     5101 ttctgagcga cgaagatgat acagacggca cggaggaaga tccctttaaa aatacattag
     5161 tacgtcacca ggatcgtttc ccctactttg cattacgcta ctttgatttg aaaaaggtgt
     5221 tcacctcttt acgtttccac atcgatcttg gtacctacca ttttgcgatc tacaaaaaga
     5281 atattgggga gcaacccgag gatcgtcatt tgacccgtaa tttgtacggg tttggtcgta
     5341 ttcaggattt tgcagaagag caccgtccgg aggagtggaa acgcttggtg cgcgacttgg
     5401 actactttga aaccggtgat aagccctata ttacgcagac tactccacac taccacattg
     5461 agaaaggtaa aattgggtta cgttttgtgc cagagggtca acatttatgg cctagccctg
     5521 aggttggggc cacccgtacg ggccgctcta agtatgcgca agacaaacgc cttactgcgg
     5581 aggcattttt gtctgttcac gagttaatgc ccatgatgtt ctattacttc ctgttacgtg
     5641 agaaatattc tgaagaagtc tccgcggaga aagtgcaagg gcgtattaaa cgcgtcatcg
     5701 aggacgtcta tgctgtgtac gacgccttcg cacgcgacga gattaacacc cgcgatgagc
     5761 ttgacgcctg tctggctgac aaggggatcc gtcgcggaca cctgccccgt cagatgattg
     5821 caatcttgtc gcaggaacat aaggacatgg aagagaaggt acgtaagaaa cttcaggaaa
     5881 tgatcgctga cacggatcac cgtttggaca tgctggaccg ccaaacagat cgcaaaattc
     5941 gcattggccg taaaaatgca ggacttccta agtccggggt cgtagcagat tggcttgtgc
     6001 gtgacatgat gcgttttcag cccgtggcga aagatactag cgggaaacca ctgaacaaca
     6061 gtaaggcgaa ctccactgaa tatcgcatgt tgcagcgcgc cttggctctg tttgggggcg
     6121 agaaggagcg tcttacgccg tacttccgtc aaatgaattt aacgggaggt aataacccgc
     6181 atcccttcct tcatgaaaca cgctgggaaa gccacaccaa tatcttgtca ttttatcgct
     6241 catatttaga agcccgtaaa gcgttccttc agtcgattgg gcgctccgat cgtgttgaga
     6301 atcaccgctt ccttttactt aaggagccca aaacagaccg tcaaacattg gttgcaggat
     6361 ggaagggtga gttccacttg cctcgtggga tcttcaccga ggctgtccgc gactgtctta
     6421 tcgaaatggg ctacgatgag gttggttcgt ataaggaagt cgggttcatg gcaaaggctg
     6481 taccactgta ctttgagcgt gcttccaaag accgtgtaca acccttttat gattatccgt
     6541 ttaacgtagg aaactctttg aaaccaaaga aaggacgttt cctgtcgaaa gaaaagcgtg
     6601 cagaggagtg ggaaagtggg aaagagcgtt ttcgcttagc aaagttgaag aaagaaatcc
     6661 tggaggccaa ggagcatcca tatcacgatt tcaaatcttg gcaaaaattc gagcgcgagc
     6721 ttcgcttagt aaagaatcag gatatcatta cttggatgat gtgtcgtgat ttgatggaag
     6781 agaataaggt tgagggctta gatacaggta ccctgtattt gaaggacatt cgcacggacg
     6841 tgcaggaaca ggggagcctg aatgttctta accgtgtgaa gccgatgcgc cttcctgttg
     6901 tcgtatatcg tgccgacagt cgcggtcacg tccataaaga acaggcccct ttggcaactg
     6961 tgtacatcga agaacgcgat actaaacttt taaaacaagg caactttaag agtttcgtta
     7021 aggaccgccg cctgaacgga ttgttctctt tcgtagatac aggtgctctt gccatggaac
     7081 agtatccgat tagtaaactg cgcgtggagt atgagcttgc caaatatcag actgctcgcg
     7141 tttgcgcttt cgaacaaaca ctggagttag aagagtcgtt acttactcgt tacccacacc
     7201 tgcctgacaa gaacttccgc aaaatgctgg agtcgtggtc ggatccgctg ttagataaat
     7261 ggcccgacct tcacgggaat gtccgcttat taattgcggt tcgcaatgct ttctcacaca
     7321 atcaatatcc aatgtatgat gagaccttgt tttcatctat ccgtaaatac gaccctagta
     7381 gcccagacgc catcgaagaa cgcatg
//
