LOCUS       sequence-428997-        3301 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression plasmid for E. coli MS 115-1 NucC, a 
            cA3-responsive nuclease known to cause abortive infection; used as a
            positive control in the Haliangium ochraceum type III CRISPR-Cas 
            system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3301)
  AUTHORS   Steens JA, Bravo JPK, Salazar CRP, Yildiz C, Amieiro AM, Kostlbacher
            S, Prinsen SHP, Andres AS, Patinios C, Bardis A, Barendregt A, 
            Scheltema RA, Ettema TJG, van der Oost J, Taylor DW, Staals RHJ
  TITLE     Type III-B CRISPR-Cas cascade of proteolytic cleavages.
  JOURNAL   Science. 2024 Feb 2;383(6682):512-519. doi: 10.1126/science.adk0378.
            Epub 2024 Feb 1.
  PUBMED    38301007
REFERENCE   2  (bases 1 to 3301)
  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.adk0378"; journalName: "Science"; date: "2024-02-2-
            2"; volume: "383"; issue: "6682"; pages: "512-519"
COMMENT     Sequence Label: pEcNucC
FEATURES             Location/Qualifiers
     source          1..3301
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(33..79)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     94..116
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     promoter        join(123..128,129..146,147..153)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     primer_bind     complement(989..1007)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1003..1050
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(join(1315..2106,2107..2175))
                     /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     1938..1957
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2176..2280)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2354..3092)
                     /direction=LEFT
                     /label=CloDF13 ori
                     /note="Plasmids containing the CloDF13 (CDF) origin of 
                     replication can be propagated in E. coli cells that contain
                     additional plasmids with compatible origins."
ORIGIN
        1 cgactcctgc attaggaaat gacaaacaac agataaaacg aaaggcccag tctttcgact
       61 gagcctttcg ttttatttga tgcctttaat taaagcggat aacaatttca cacaggaggc
      121 cgtttacact ttatgcttcc ggctcgtata atgtgtgggg aaaccacaac ggtttccctc
      181 tagaaataat tttgtttaac tataagaagg agatatacat atgtcagatt ggtcgctttc
      241 tcaactattt gcatcgttac atgaagacat tcagcttcga ctgggaactg cacgtaaagc
      301 tttccagcat cccggggcta aaggggatgc cagtgaagga gtatggattg aaatgcttga
      361 tacatatctt cccaaacgct atcaggcagc taatgctttt gtagttgata gtttgggtaa
      421 ttttagtgat cagattgatg tggttgtttt tgacaggcag tactcaccat tcattttcaa
      481 atttaatgaa caaataattg ttccagcaga aagtgtctat gcggtttttg aagcaaagca
      541 atctgcgtca gcagatcttg ttgcttatgc acagcgaaaa gttgcaagtg ttcgcagact
      601 tcatcgtacc agccttccaa ttccgcatgc tggcgggaca tatccagcaa aaccactcat
      661 tccaattttg ggagggttac ttacttttga aagtgactgg agtcctgcat tggggatgtc
      721 gtttgacaaa gccctcaatg gggatttatc tgatggacga cttgatatgg gctgtgttgc
      781 atcgcatggg catttttact tcaataacat tgatagcaag ttcaattttg aacatggaaa
      841 taaaccagct accgcttttc ttttcaggct tattgcgcaa cttcaattta gtggcactgt
      901 gccaatgatt gacattgatg cctatggaaa atggttggct aattgatgcc gtgggacagc
      961 ccgtcttaat taacctaggc tgctgccacc gctgagcaat aactagcata accccttggg
     1021 gcctctaaac gggtcttgag tggttttttg ctgaaacctc aggcatttga gaagcacacg
     1081 gtcacactgc ttccggtagt caataaaccg gtaaaccagc aatagacata agcggctatt
     1141 taacgaccct gccctgaacc gacgaccggg tcatcgtggc cggatcttgc ggcccctcgg
     1201 cttgaacgaa ttgttagaca gattatcaaa aaggatcttc acctagatcc ttttaaatta
     1261 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     1321 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     1381 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     1441 tgcaatgata ccgcgggacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     1501 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     1561 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     1621 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     1681 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     1741 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     1801 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     1861 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     1921 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     1981 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     2041 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     2101 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     2161 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     2221 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     2281 cacatttccc cgaaaagtgc cacctgtagc tagctcactc ggtcgctacg ctccgggcgt
     2341 gagactgcgg cgggcgctgc ggacacatac aaagttaccc acagattccg tggataagca
     2401 ggggactaac atgtgaggca aaacagcagg gccgcgccgg tggcgttttt ccataggctc
     2461 cgccctcctg ccagagttca cataaacaga cgcttttccg gtgcatctgt gggagccgtg
     2521 aggctcaacc atgaatctga cagtacgggc gaaacccgac aggacttaaa gatccccacc
     2581 gtttccggcg ggtcgctccc tcttgcgctc tcctgttccg accctgccgt ttaccggata
     2641 cctgttccgc ctttctccct tacgggaagt gtggcgcttt ctcatagctc acacactggt
     2701 atctcggctc ggtgtaggtc gttcgctcca agctgggctg taagcaagaa ctccccgttc
     2761 agcccgactg ctgcgcctta tccggtaact gttcacttga gtccaacccg gaaaagcacg
     2821 gtaaaacgcc actggcagca gccattggta actgggagtt cgcagaggat ttgtttagct
     2881 aaacacgcgg ttgctcttga agtgtgcgcc aaagtccggc tacactggaa ggacagattt
     2941 ggttgctgtg ctctgcgaaa gccagttacc acggttaagc agttccccaa ctgacttaac
     3001 cttcgatcaa accacctccc caggtggttt tttcgtttac agggcaaaag attacgcgca
     3061 gaaaaaaagg atctcaagaa gatcctttga tcttttctac tgaaccgctc tagatttcag
     3121 tgcaatttat ctcttcaaat gtagcacctg aagtcagccc catacgatat aagttgtaat
     3181 tctcatgtta gtcatgcccc gcgcccaccg gaaggagctg actgggttga aggctctcaa
     3241 gggcatcggt cgagatcccg gtgcctaatg agtgagctaa cttacattaa ttgcgttgcg
     3301 c
//
