LOCUS       sequence-218327-        3046 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Vector for anti-CRISPR expression with pTet.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3046)
  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 3046)
  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: pKEW270-pTet-empty
FEATURES             Location/Qualifiers
     source          1..3046
                     /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      1123..1150
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     rep_origin      complement(1276..1864)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1356..1375)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2035..2826,2827..2895))
                     /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     2658..2677
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2896..3000)
                     /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 tagagaaaag aattcaactg caggcactgc ccatggacct cggtaccgaa
      781 tagctagccg gtaatgcatt cgctagagct cctaaagcat gcgacctgca accggtctgt
      841 cacgtacgtc gccaccgtcg acgtcgttcg taagtagcct agataaataa aataatcagt
      901 taaccgcgag ccccatgcga gagtagggaa ctgccaggca tttcagccaa aaaacttaag
      961 accgccggtc ttgtccacta ccttgcagta atgcggtgga caggatcggc ggttttcttt
     1021 tctcttctca accgccggga gcggatttga acgttgcgaa gcaacggccc ggagggtggc
     1081 gggcaggacg cccgccataa actgccaggc atcaaattaa gcagaaggcc atcctgacgg
     1141 atggcctttt tgcgtttcta caaactctgc ggtaatacgg ttatccacag aatcagggga
     1201 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     1261 cgcgttgctg gcgtttttcc acaggctccg cccccctgac gagcatcaca aaaatcgacg
     1321 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     1381 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     1441 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     1501 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     1561 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     1621 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     1681 cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct
     1741 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     1801 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     1861 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     1921 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     1981 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     2041 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     2101 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     2161 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     2221 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     2281 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     2341 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     2401 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     2461 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     2521 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     2581 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     2641 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     2701 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     2761 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     2821 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     2881 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     2941 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     3001 cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attatt
//
