LOCUS       sequence-178223-        3264 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses csx28 from P. buccae ATCC 33574 in bacteria. Expression 
            driven from the Lac promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3264)
  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 3264)
  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: pPbcsx28
FEATURES             Location/Qualifiers
     source          1..3264
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     36..55
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     249..271
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     263..280
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     264..280
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(join(850..855,856..873,874..879))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(928..944)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(928..944)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(941..963)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    952..968
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(976..982,983..1000,1001..1006))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1021..1042
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1159..1176)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1330..1918)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1410..1429)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2089..2880,2881..2949))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2712..2731
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2950..3054)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3122..3140
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3178..3200)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga ttgtactgag
       61 agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag
      121 gcgccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc
      181 gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc
      241 agggttttcc cagtcacgac gttgtaaaac gacggccagt gaattcgagc tcggtacccg
      301 gttaaaatct taaatctcgt tttttcagat tgattgcttg tctgatttct atcgagaggt
      361 tttggtgaat ctttttcatt cgttctacag actctctatt agttattctt attgtttctt
      421 ggggattgtt ttgggcggaa agcatgaaac cataaacaat atttctatat tcggaaatta
      481 gtgacagagc ctcttttgac ataaaaatac cacatccttc gttatatatt tctttggaaa
      541 gctcttttat aaatttgcgt atgttttctt ttctaaaata ataggttgct tcttgcttac
      601 catcttttgt tttttcccaa atcaatatgc tgtcttcatt ttcagtattt gtagtaaatc
      661 tgagtagttt atacaggctt ttatgtgctt gtaacacagc ttggtatttg gcatattcaa
      721 tattctcaac acttttttgg tgtagttgct tgatagcata gtaataggcc acgtatgctg
      781 ctatgaaggt acatattata gagaatgcct ctttcgctaa ttccatataa tccatgataa
      841 agacctccta acatacgagc cggaagcata aagtgtaaag gatcctctag agtcgacctg
      901 caggcatgca agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
      961 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     1021 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     1081 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     1141 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     1201 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     1261 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     1321 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     1381 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     1441 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     1501 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     1561 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     1621 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     1681 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     1741 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     1801 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     1861 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     1921 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     1981 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     2041 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     2101 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     2161 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     2221 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     2281 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     2341 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     2401 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     2461 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     2521 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     2581 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     2641 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     2701 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     2761 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     2821 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     2881 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     2941 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     3001 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     3061 tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa
     3121 aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct
     3181 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag
     3241 acaagcccgt cagggcgcgt cagc
//
