LOCUS       sequence-225592-        2974 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Vector for anti-CRISPR expression with pTet.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 2974)
  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 2974)
  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..2974
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        280..335
                     /gene="tetR"
                     /label=tetR/tetA promoters
                     /note="overlapping promoters for bacterial tetR and tetA"
     protein_bind    281..299
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             351..977
                     /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     complement(427..446)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     promoter        1063..1167
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1168..1236,1237..2028)
                     /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(1386..1405)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2199..2787
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2688..2707
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2913..2940)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
ORIGIN
        1 aaaaccgccg atcctgtcca ccgcattact gcaaggtagt ggacaagacc ggcggtctta
       61 agttttttgg ctgaaatgcc tggcagttcc ctactctcgc atggggctcg cggttaactg
      121 attattttat ttatctaggc tacttacgaa cgacgtcgac ggtggcgacg tacgtgacag
      181 accggttgca ggtcgcatgc tttaggagct ctagcgaatg cattaccggc tagctattcg
      241 gtaccgaggt ccatgggcag tgcctgcagt tgaattcttt tctctatcac tgatagggag
      301 tggtaaaata actctatcaa cgatagagtg tcaacaaaaa ttaggaatta atgatgtcta
      361 gattagataa aagtaaagtg attaacagcg cattagagct gcttaatgag gtcggaatcg
      421 aaggtttaac aacccgtaaa ctcgcccaga agctaggtgt agagcagcct acattgtatt
      481 ggcatgtaaa aaataagcgg gctttgctcg acgccttagc cattgagatg ttagataggc
      541 accatactca cttttgccct ttagaagggg aaagctggca agatttttta cgtaataacg
      601 ctaaaagttt tagatgtgct ttactaagtc atcgcgatgg agcaaaagta catttaggta
      661 cacggcctac agaaaaacag tatgaaactc tcgaaaatca attagccttt ttatgccaac
      721 aaggtttttc actagagaat gcattatatg cactcagcgc tgtggggcat tttactttag
      781 gttgcgtatt ggaagatcaa gagcatcaag tcgctaaaga agaaagggaa acacctacta
      841 ctgatagtat gccgccatta ttacgacaag ctatcgaatt atttgatcac caaggtgcag
      901 agccagcctt cttattcggc cttgaattga tcatatgcgg attagaaaaa caacttaaat
      961 gtgaaagtgg gtcttaagac gtcctatcaa ggctaactcg agtaaggtat atccacaata
     1021 atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     1081 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     1141 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     1201 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     1261 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     1321 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     1381 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     1441 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     1501 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     1561 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     1621 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     1681 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     1741 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     1801 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     1861 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     1921 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     1981 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     2041 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     2101 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     2161 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     2221 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     2281 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     2341 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     2401 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     2461 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     2521 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     2581 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     2641 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     2701 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     2761 tcgtcagggg ggcggagcct gtggaaaaac gccagcaacg cggccttttt acggttcctg
     2821 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     2881 aaccgtatta ccgcagagtt tgtagaaacg caaaaaggct atccgtcagg atggccttct
     2941 gcttaatttg atgcctggca gtttatggcg ggcg
//
