LOCUS       sequence-178224-        7808 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression for pbCas13b, driven by the lac promoter, and 
            DR-spacer-DR sequence driven by js23119. New spacer sequences can be
            cloned in between the DRs by digesting the plasmid with BsaI..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7808)
  AUTHORS   Smargon AA, Cox DB, Pyzocha NK, Zheng K, Slaymaker IM, Gootenberg 
            JS, Abudayyeh OA, Essletzbichler P, Shmakov S, Makarova KS, Koonin 
            EV, Zhang F
  TITLE     Cas13b Is a Type VI-B CRISPR-Associated RNA-Guided RNase 
            Differentially Regulated by Accessory Proteins Csx27 and Csx28.
  JOURNAL   Mol Cell. 2017 Feb 16;65(4):618-630.e7. doi: 
            10.1016/j.molcel.2016.12.023. Epub 2017 Jan 5.
  PUBMED    28065598
REFERENCE   2  (bases 1 to 7808)
  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"; doi: 
            "10.1016/j.molcel.2016.12.023"; journalName: "Mol Cell"; date: 
            "2017-02-16- 16"; volume: "65"; issue: "4"; pages: "618-630.e7"
COMMENT     Sequence Label: pPbcas13b
FEATURES             Location/Qualifiers
     source          1..7808
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(join(3303..3308,3309..3326,3327..3332))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     promoter        complement(join(3482..3487,3488..3504,3505..3510))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     rep_origin      3622..4167
                     /direction=RIGHT
                     /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     4068..4087
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4284..4301
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        4693..4795
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             4796..5455
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(4862..4881)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     7308..7327
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        complement(7677..7711)
                     /label=J23119 promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119)"
ORIGIN
        1 gacagaaggg aaaagagtaa attttttaac ttcagcaaag agtgtggcat catacatggg
       61 atactggttg tgggaaaaag catttctcac tgcaatcaaa ctattaacgt aattctttaa
      121 ttccggtcta ttagctttgc actgcaacca cctttcaagc atatttctga agctatctgt
      181 aggtagtgtg gaatatttgt ctatgagttt cttttccagt cctaatgtca tctcaaagat
      241 actaatcctt gtagtctgat attttataag ctcaaggtct acactcaatt tgctgatagg
      301 atgttcctct aagttcaagt ctgtagtttc tgcaaatgag aatagcccat tgagtcttcg
      361 gtcttttacc aatgctttga aattgccctg tttcaggact ttggtttccg tttcttcgat
      421 ataaaaggtt gctaaaggtc tttcctttaa tatatttcct ttattgtctg tttcgtatgt
      481 tttaacaggc aacttcatgg gcatgattct atttagaata ttgttgcttt cctcatttgc
      541 cgtgtttgta tcgatgtccc tcagatgtat ttctccaatc ttcaatcctt ccacagtagc
      601 catattgaat agttctttga acatcaacca tgtgacgata tcctgattct tgataagtcg
      661 cagttcgcgt tcgaactttt tccatgataa aaaatcaaga taagctaagt ctgtcttttt
      721 cttttcagac ggataatttt tgaatagctc tttattcttt tgccacaatt cgacacgctc
      781 ttttttatct aagaattgcc ttttttttgg tttaagtctg ttgccgatat tgaacggata
      841 atcgtagaaa ggctgtacat tgtctttata ttcttctgca aagtataagg gaatagcttt
      901 tgcaacaaac cctactcggt caaaagatag tatttggtcg tatattcttt ttgaattatt
      961 atgtttctca aaccattccc ttattggttg ggtaaatatt ccacggggga gattgaaact
     1021 gtttttccat cctgtaacta atgtgttgcg gttggtcttc tgtactttaa gtatgagaaa
     1081 atgctgatat tgtttccagt tttgtggttt taagccctta agatactttt tcctcgcttc
     1141 caaatagttt cggtagaaag acagaatgtt agtttgatgt tcccattgtg tttctgcaag
     1201 gaaaggatgc ggattgtcat tccccacaag attcatttga ttaaaatatg ccttaaggcg
     1261 gaagttctca ctgccgaaaa gggcaagcgc acgctgaagc attcgatatt cggtgctgtt
     1321 agccttgcta ttgtttattg gaatgttatt ttggtctttt tgaacaggtt ggaagcgcat
     1381 catgtcattt accaaccaat ctgcaatttt tcccgatttt agcagaccgg cattcctttt
     1441 gccaatcctt atcttctgat ttgtctgctt gcagagcaag tcaagtcttc tttgggtgtc
     1501 gtctatcatt tccccaattt tgcgctcagc ctcttttccc atatcttttt gtctgccttt
     1561 caatatggaa atcatttgtt ttggcaaatg gccttgtagt atatttgtat tttgaagccg
     1621 acgggttaag tctgcaatgc tatttatctc attattggca aaagcatcgt agatagcgta
     1681 tatgttgctt atttcttttc ttataattcc ctctactttg tctgcgcttt cttttctgga
     1741 gtaatccttg gtcaataaca gataataaaa catcatcggc aacaattcgt ggacacttaa
     1801 aaatgcttct gctgtaaatt gtgttccgag attaaattta cttcttccat tgcatgtttt
     1861 gtcggtctga agagatggaa agaggttgtt gtgggcactg caaaacttaa tgcctatctt
     1921 ttcgttctct aaatggtaat gtggagcagt cttagatatg taaggctctt gcgatgtctc
     1981 aaaatgatct aagtctttca ctaattttct ccattcttct ggttggtttt gtggtgcaaa
     2041 gtcctgaatg cgcgcaaagc catatagatg atgtgtgaga tggcgaactt catcttcgtc
     2101 accgatacgt ttgttataga tagagaaatg ataggtacct aaatctattt ggaagcggag
     2161 ttgttcgaat atctcattta aatcgaaata tctcaagaca aaatatggaa agcggtcttg
     2221 atgccttaca agcgtgttct taaatggttc ttcttcagca ttatagtcgt cagagaaaat
     2281 atcgaaaggt actttaaaac tttccctatc tttttctcgt aaccgttcat acagcgattt
     2341 gggacagcga accaattcgt taagcatatc aagttgcatc cagtcttttg tctgtacatt
     2401 ctctaatttt agcttaggca aagatatacg gcttctgcaa aatacttcgt ttgtcatttg
     2461 ctcacgcaag tttctgccat ctttaaatcc ctttagcttt ttctgcatcc aaatagcgtc
     2521 ttttttatca agaaataaag agacgaagaa cagtaagcct gctatggtca tattgccttc
     2581 tttatctgca aaatggtaat gaaagttagg actgtctatg atgttttttg ttcgtcccac
     2641 ttgttttttc ctgttgagat gtgtaaaatc tctttgcatg tctatattct catgatgcat
     2701 atagtctctc tttactaacc gcacattggc atcgaaaacc ttatacatgt ttttcaaaag
     2761 gcttgtttca aatatcggtt ttggggattc ttcagaatac ttgtaatggg agtagtagtt
     2821 cctgagaacc tgaagttttt ccaagaaaat gaatagaaca ttcttcaagt tgttaagtga
     2881 tagggcttcg gattgttcct tttcccgttg ctccttattg ttgggacttt tagagtttgt
     2941 catttcataa gctgcagctt ccagaaaagg aaagtgtttc atgatcaagt ctctcagccg
     3001 aacttttttg tcgagttttt tcgcttgctc gttccagctg cctttaatgc ccatcatata
     3061 tccatcgtct tttagctctg ctatttccaa acgtctgttg atgtgattaa tggtggtata
     3121 gacattgtgg cgcgccaagt taagaaaggc agcccagaaa tgcttatctg tcaattcata
     3181 ataaattggt tcaggtttct ctttgttttc cattattgtt atatatttag gaaaggcaaa
     3241 tatagcattt ttctttctat ctacaaataa tttatcttgt ttttgcatga taaagacctc
     3301 ctaacatacg agccggaagc ataaagtgta aaatcccgca agaggcccgg cagtaccggc
     3361 ataaccaagc ctatgcctac agcatccagg gtgacggtgc cgaggatgac gatgagcgca
     3421 ttgttagatt tcatacacgg tgcctgactg cgttagcaat ttaactgtga taaactaccg
     3481 cattaaagct tatcgatgat aagctgtcaa acatgagaat tacaacttat atcgtatggg
     3541 gctgacttca ggtgctacat ttgaagagat aaattgcact gaaatctaga aatattttat
     3601 ctgattaata agatgatctt cttgagatcg ttttggtctg cgcgtaatct cttgctctga
     3661 aaacgaaaaa accgccttgc agggcggttt ttcgaaggtt ctctgagcta ccaactcttt
     3721 gaaccgaggt aactggcttg gaggagcgca gtcaccaaaa cttgtccttt cagtttagcc
     3781 ttaaccggcg catgacttca agactaactc ctctaaatca attaccagtg gctgctgcca
     3841 gtggtgcttt tgcatgtctt tccgggttgg actcaagacg atagttaccg gataaggcgc
     3901 agcggtcgga ctgaacgggg ggttcgtgca tacagtccag cttggagcga actgcctacc
     3961 cggaactgag tgtcaggcgt ggaatgagac aaacgcggcc ataacagcgg aatgacaccg
     4021 gtaaaccgaa aggcaggaac aggagagcgc acgagggagc cgccaggggg aaacgcctgg
     4081 tatctttata gtcctgtcgg gtttcgccac cactgatttg agcgtcagat ttcgtgatgc
     4141 ttgtcagggg ggcggagcct atggaaaaac ggctttgccg cggccctctc acttccctgt
     4201 taagtatctt cctggcatct tccaggaaat ctccgccccg ttcgtaagcc atttccgctc
     4261 gccgcagtcg aacgaccgag cgtagcgagt cagtgagcga ggaagcggaa tatatcctgt
     4321 atcacatatt ctgctgacgc accggtgcag ccttttttct cctgccacat gaagcacttc
     4381 actgacaccc tcatcagtgc caacatagta agccagtata cactccgcta gcgctgatgt
     4441 ccggcggtgc ttttgccgtt acgcaccacc ccgtcagtag ctgaacagga gggacagctg
     4501 atagaaacag aagccactgg agcacctcaa aaacaccatc atacactaaa tcagtaagtt
     4561 ggcagcatca cccgacgcac tttgcgccga ataaatacct gtgacggaag atcacttcgc
     4621 agaataaata aatcctggtg tccctgttga taccgggaag ccctgggcca acttttggcg
     4681 aaaatgagac gttgatcggc acgtaagagg ttccaacttt caccataatg aaataagatc
     4741 actaccgggc gtattttttg agttatcgag attttcagga gctaaggaag ctaaaatgga
     4801 gaaaaaaatc actggatata ccaccgttga tatatcccaa tggcatcgta aagaacattt
     4861 tgaggcattt cagtcagttg ctcaatgtac ctataaccag accgttcagc tggatattac
     4921 ggccttttta aagaccgtaa agaaaaataa gcacaagttt tatccggcct ttattcacat
     4981 tcttgcccgc ctgatgaatg ctcatccgga attccgtatg gcaatgaaag acggtgagct
     5041 ggtgatatgg gatagtgttc acccttgtta caccgttttc catgagcaaa ctgaaacgtt
     5101 ttcatcgctc tggagtgaat accacgacga tttccggcag tttctacaca tatattcgca
     5161 agatgtggcg tgttacggtg aaaacctggc ctatttccct aaagggttta ttgagaatat
     5221 gtttttcgtc tcagccaatc cctgggtgag tttcaccagt tttgatttaa acgtggccaa
     5281 tatggacaac ttcttcgccc ccgttttcac catgggcaaa tattatacgc aaggcgacaa
     5341 ggtgctgatg ccgctggcga ttcaggttca tcatgccgtt tgtgatggct tccatgtcgg
     5401 cagaatgctt aatgaattac aacagtactg cgatgagtgg cagggcgggg cgtaattttt
     5461 ttaaggcagt tattggtgcc cttaaacgcc tggttgctac gcctgaataa gtgataataa
     5521 gcggatgaat ggcagaaatt cgaaagcaaa ttcgacccgg tcgtcggttc agggcagggt
     5581 cgttaaatag ccgcttatgt ctattgctgg tttaccggtt tattgactac cggaagcagt
     5641 gtgaccgtgt gcttctcaaa tgcctgaggc cagtttgctc aggctctccc cgtggaggta
     5701 ataattgacg atatgatcat ttattctgcc tcccagagcc tgataaaaac ggttagcgct
     5761 tcgttaatac agatgtaggt gttccacagg gtagccagca gcatcctgcg atgcagatcc
     5821 ggaacataat ggtgcagggc gcttgtttcg gcgtgggtat ggtggcaggc cccgtggccg
     5881 ggggactgtt gggcgctgcc ggcacctgtc ctacgagttg catgataaag aagacagtca
     5941 taagtgcggc gacgatagtc atgccccgcg cccaccggaa ggagctaccg gcagcggtgc
     6001 ggactgttgt aactcagaat aagaaatgag gccgctcatg gcgttgactc tcagtcatag
     6061 tatcgtggta tcaccggttg gttccactct ctgttgcggg caacttcagc agcacgtagg
     6121 ggacttccgc gtttccagac tttacgaaac acggaaaccg aagaccattc atgttgttgc
     6181 tcaggtcgca gacgttttgc agcagcagtc gcttcacgtt cgctcgcgta tcggtgattc
     6241 attctgctaa ccagtaaggc aaccccgcca gcctagccgg gtcctcaacg acaggagcac
     6301 gatcatgcgc acccgtggcc aggacccaac gctgcccgag atgcgccgcg tgcggctgct
     6361 ggagatggcg gacgcgatgg atatgttctg ccaagggttg gtttgcgcat tcacagttct
     6421 ccgcaagaat tgattggctc caattcttgg agtggtgaat ccgttagcga ggtgccgccg
     6481 gcttccattc aggtcgaggt ggcccggctc catgcaccgc gacgcaacgc ggggaggcag
     6541 acaaggtata gggcggcgcc tacaatccat gccaacccgt tccatgtgct cgccgaggcg
     6601 gcataaatcg ccgtgacgat cagcggtcca atgatcgaag ttaggctggt aagagccgcg
     6661 agcgatcctt gaagctgtcc ctgatggtcg tcatctacct gcctggacag catggcctgc
     6721 aacgcgggca tcccgatgcc gccggaagcg agaagaatca taatggggaa ggccatccag
     6781 cctcgcgtcg cgaacgccag caagacgtag cccagcgcgt cggccgccat gccggcgata
     6841 atggcctgct tctcgccgaa acgtttggtg gcgggaccag tgacgaaggc ttgagcgagg
     6901 gcgtgcaaga ttccgaatac cgcaagcgac aggccgatca tcgtcgcgct ccagcgaaag
     6961 cggtcctcgc cgaaaatgac ccagagcgct gccggcacct gtcctacgag ttgcatgata
     7021 aagaagacag tcataagtgc ggcgacgata gtcatgcccc gcgcccaccg gaaggagctg
     7081 actgggttga aggctctcaa gggcatcggt cgacgctctc ccttatgcga ctcctgcatt
     7141 aggaagcagc ccagtagtag gttgaggccg ttgagcaccg ccgccgcaag gaatggtgca
     7201 tgcaaggaga tggcgcccaa cagtcccccg gccacggggc ctgccaccat acccacgccg
     7261 aaacaagcgc tcatgagccc gaagtggcga gcccgatctt ccccatcggt gatgtcggcg
     7321 atataggcgc cagcaaccgc acctgtggcg ccggtgatgc cggccacgat gcgtccggcg
     7381 tagaggatcc acaggacggg tgtggtcgcc atgatcgcgt agtcgatagt ggctccaagt
     7441 agcgaagcga gcaggactgg gcggcggcca aagcggtcgg acagtgctcc gagaacgggt
     7501 gcgcatagaa attgcatcaa cgcatatagc gctagcagca cgccatagtg actggcgatg
     7561 ctgtcggaat ggacgatgtt gtttttacct ttcaaaaaga aggcagatgc aactgagacc
     7621 ttccgccatg gtctcgttgt ttttaccttt caaaaagaag gcagatgcaa ctcagcgcta
     7681 gcattatacc taggactgag ctagctgtca atttagtttt tattttctga tttttcaatt
     7741 tcttttatgg cttttcctac tatttccagt aactgtggcg ctatatttag ctcaatcttt
     7801 tttgtatc
//
