LOCUS       sequence-218332-        3198 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses AcrVA5 in bacteria, under TetR control..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3198)
  AUTHORS   Watters KE, Fellmann C, Bai HB, Ren SM, Doudna JA
  TITLE     Systematic discovery of natural CRISPR-Cas12a inhibitors.
  JOURNAL   Science. 2018 Oct 12;362(6411):236-239. doi: 
            10.1126/science.aau5138. Epub 2018 Sep 6.
  PUBMED    30190307
REFERENCE   2  (bases 1 to 3198)
  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: 
            "10.1126/science.aau5138"; journalName: "Science"; date: 
            "2018-10-12- 12"; volume: "362"; issue: "6411"; pages: "236-239"
COMMENT     Sequence Label: pKEW303-AcrVA5
FEATURES             Location/Qualifiers
     source          1..3198
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(40..666)
                     /codon_start=1
                     /gene="tetR"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MMSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWH
                     VKNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGT
                     RPTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPT
                     TDSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     571..590
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     promoter        682..737
                     /gene="tetR"
                     /label=tetR/tetA promoters
                     /note="overlapping promoters for bacterial tetR and tetA"
     protein_bind    718..736
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     terminator      1275..1302
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     rep_origin      complement(1428..2016)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1508..1527)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2187..2978,2979..3047))
                     /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     2810..2829
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3048..3152)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtggatatac cttactcgag ttagccttga taggacgtct taagacccac tttcacattt
       61 aagttgtttt tctaatccgc atatgatcaa ttcaaggccg aataagaagg ctggctctgc
      121 accttggtga tcaaataatt cgatagcttg tcgtaataat ggcggcatac tatcagtagt
      181 aggtgtttcc ctttcttctt tagcgacttg atgctcttga tcttccaata cgcaacctaa
      241 agtaaaatgc cccacagcgc tgagtgcata taatgcattc tctagtgaaa aaccttgttg
      301 gcataaaaag gctaattgat tttcgagagt ttcatactgt ttttctgtag gccgtgtacc
      361 taaatgtact tttgctccat cgcgatgact tagtaaagca catctaaaac ttttagcgtt
      421 attacgtaaa aaatcttgcc agctttcccc ttctaaaggg caaaagtgag tatggtgcct
      481 atctaacatc tcaatggcta aggcgtcgag caaagcccgc ttatttttta catgccaata
      541 caatgtaggc tgctctacac ctagcttctg ggcgagttta cgggttgtta aaccttcgat
      601 tccgacctca ttaagcagct ctaatgcgct gttaatcact ttacttttat ctaatctaga
      661 catcattaat tcctaatttt tgttgacact ctatcgttga tagagttatt ttaccactcc
      721 ctatcagtga tagagaaagt actaaggtgc aggagttaag aaaatgaaaa tcgaactatc
      781 aggcggatat atctgctaca gcattgaaga agatgaagtt accattgaca tggtagaggt
      841 taccaccaaa cgtcaaggca ttggcagtca gctaattgat atggttaagg atgtagcccg
      901 tgaagttggc ttgccaattg gtctatatgc ctacccccaa gatgacagca ttagccaaga
      961 agacttgatt gagttttatt tttctaatga ctttgagtat gacccagatg atgtagatgg
     1021 tcgcttgatg agatggagct aaaactcaga agtgaaacgc cgccccatgc gagagtaggg
     1081 aactgccagg catttcagcc aaaaaactta agaccgccgg tcttgtccac taccttgcag
     1141 taatgcggtg gacaggatcg gcggttttct tttctcttct caaccgccgg gagcggattt
     1201 gaacgttgcg aagcaacggc cggaagggtg gcgggcagga cgcccgccat aaactgccag
     1261 gcatcaaatt aagcagaagg ccatcctgac ggatggcctt tttgcgtttc tacaaactct
     1321 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     1381 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccacaggctc
     1441 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     1501 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     1561 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     1621 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     1681 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     1741 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     1801 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     1861 actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     1921 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     1981 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     2041 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     2101 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     2161 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     2221 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     2281 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     2341 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     2401 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     2461 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     2521 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     2581 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     2641 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     2701 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     2761 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     2821 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     2881 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     2941 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     3001 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     3061 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     3121 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     3181 gtctaagaaa ccattatt
//
