LOCUS       sequence-174207-        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     449..468
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     702..719
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    836..857
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(872..877,878..895,896..902)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    910..926
                     /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     915..937
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     934..950
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     934..950
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        join(999..1004,1005..1022,1023..1028)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(1598..1615)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1598..1614)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1607..1629)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(1823..1842)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1942..1964
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2002..2020)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2088..2192
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2193..2261,2262..3053)
                     /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(2411..2430)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(3224..3264,1..548)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
       61 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
      121 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
      181 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
      241 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
      301 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
      361 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
      421 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
      481 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
      541 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
      601 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
      661 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
      721 aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat
      781 gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg
      841 tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt
      901 tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg
      961 ccaagcttgc atgcctgcag gtcgactcta gaggatcctt tacactttat gcttccggct
     1021 cgtatgttag gaggtcttta tcatggatta tatggaatta gcgaaagagg cattctctat
     1081 aatatgtacc ttcatagcag catacgtggc ctattactat gctatcaagc aactacacca
     1141 aaaaagtgtt gagaatattg aatatgccaa ataccaagct gtgttacaag cacataaaag
     1201 cctgtataaa ctactcagat ttactacaaa tactgaaaat gaagacagca tattgatttg
     1261 ggaaaaaaca aaagatggta agcaagaagc aacctattat tttagaaaag aaaacatacg
     1321 caaatttata aaagagcttt ccaaagaaat atataacgaa ggatgtggta tttttatgtc
     1381 aaaagaggct ctgtcactaa tttccgaata tagaaatatt gtttatggtt tcatgctttc
     1441 cgcccaaaac aatccccaag aaacaataag aataactaat agagagtctg tagaacgaat
     1501 gaaaaagatt caccaaaacc tctcgataga aatcagacaa gcaatcaatc tgaaaaaacg
     1561 agatttaaga ttttaaccgg gtaccgagct cgaattcact ggccgtcgtt ttacaacgtc
     1621 gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg
     1681 ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc
     1741 tgaatggcga atggcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac
     1801 accgcatatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc
     1861 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt
     1921 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac
     1981 cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga
     2041 taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     2101 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     2161 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     2221 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     2281 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     2341 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     2401 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     2461 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     2521 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     2581 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     2641 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     2701 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     2761 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     2821 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     2881 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     2941 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     3001 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     3061 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     3121 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     3181 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     3241 tgcgcgtaat ctgctgcttg caaa
//
