LOCUS       sequence-174196-        3336 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses csx27 from B. zoohelcum ATCC 43767 in bacteria. Expression
            driven from the Lac promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3336)
  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 3336)
  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: pBZcsx27
FEATURES             Location/Qualifiers
     source          1..3336
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     475..494
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     728..745
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    862..883
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(898..903,904..921,922..928)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    936..952
                     /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)."
     primer_bind     941..963
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     960..976
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     960..976
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        join(1025..1030,1031..1048,1049..1054)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(1696..1713)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1696..1712)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1705..1727)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(1921..1940)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2040..2062
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2100..2118)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2186..2290
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2291..2359,2360..3151)
                     /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     complement(2509..2528)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(3322..3336,1..574)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
       61 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
      121 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
      181 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
      241 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
      301 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
      361 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
      421 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
      481 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
      541 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
      601 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
      661 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
      721 cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct
      781 ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc
      841 gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt
      901 acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac
      961 aggaaacagc tatgaccatg attacgccaa gcttgcatgc ctgcaggtcg actctagagg
     1021 atcctttaca ctttatgctt ccggctcgta tgttaggagg tctttatcat gattgagctg
     1081 ataaattttt cagcggaata tgtttttggg aaagccccac tcattacagc aggaaacctt
     1141 agcgacggtc tgttgttttt tgccattgcc ctatttatag gatgcttgat tcatatcatc
     1201 acattttggt tgataaagag aaagtggtat aagaaattgg cttgtacgcc tatggacaaa
     1261 atgaaaaatg acgagtacat cactgaaatt cttcctgatt tgaaagaaat ttacagaaca
     1321 aataaaggaa tttcggtaac agaaaatatc agtaatggaa gtgtctttga tacagcgtat
     1381 tattatttag aggcacaagg caaaatatcc caagcaaaaa attttcagag tatttatttc
     1441 ttgtttcgga acatcgtaac gctttcgtta tttgtacttc ccgtgagtgt tatcttttta
     1501 ttagcatcct tttttatgaa tgattgttcg ctgagtgaga aaatcattac aattattata
     1561 ggaaccttcg ttctaggtgg attatcatcg gttatagccc aatggtttcg ggtaaaaatg
     1621 acagacagaa ttttcggact ttactatgca gaattaacac acaataaaaa ataaccgggt
     1681 accgagctcg aattcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt
     1741 tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga
     1801 ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggcgcctgat
     1861 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatatggt gcactctcag
     1921 tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga
     1981 cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc
     2041 cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg
     2101 cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc
     2161 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     2221 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     2281 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     2341 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     2401 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     2461 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     2521 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     2581 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     2641 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     2701 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     2761 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     2821 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     2881 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     2941 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     3001 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     3061 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     3121 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     3181 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     3241 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     3301 agaaaagatc aaaggatctt cttgagatcc tttttt
//
