LOCUS       sequence-417587-        5604 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression plasmid for SAVED-CHAT and PCaspase C-terminal 
            fragment (aa 154-666) from Haliangium ochraceum.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5604)
  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 5604)
  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: pSAVED-CHAT_PCaspase-C-term
FEATURES             Location/Qualifiers
     source          1..5604
                     /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(3409..3427)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3423..3470
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(join(3618..4409,4410..4478))
                     /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     4241..4260
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4479..4583)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4657..5395)
                     /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 atgactcacc atgctgatca
      241 tgactgccag ccgcacaggg agagcgttca gccggatgaa atccagtgta tcctggttct
      301 ggacctgagc atcgataacg caatcaccgc atgtagcgtt accccgcatc tgccccgtgc
      361 tgcacgtcgt gttgaactac atctgaacga ctttggtgca gaacgtgcac cgtacggggg
      421 tgcaagcgat cgtcgtacct ggcgttgttg gatgcaggca gttgatgcaa tgctggctga
      481 cgcacgtgca cagctgggtg cagaagttga atttactcat tattatctgg caggtcgtgc
      541 agcactcccg gtttttgcat atctgggtct gagactgggt aaacaggcaa acatcactac
      601 cgttaaccgt cgtgatgacg gttgttggga tgttgttccg tgccagcgtc cggcttcgag
      661 cgcagcatcg ggggtttcgc ccagcgcacg tttttttgac gaagttcgtg gtctggacac
      721 cgatgagcgt agcagcgaaa gcggtatggt tgcagtttgg gtaagcaccc agcgtgatgt
      781 tgatcgtggg ctgctgcgtg catttgcaag ggcaagaggt gaccgtgacc tggcaggtat
      841 cgttagcctg cgtgcacgtc cggctgcagg tgatgacacc ggtgatatgc gtctcctgga
      901 aggtgcagac ggccccgatg ctgcacgtga actggttaac tgttttcgta gcatcccgaa
      961 ccagtatccg cgtagcagcg gcctgatggt ttttgttagc ggcccggtta ccctggcagc
     1021 aatggttggg cgtgcaatca acccgagaat ccatggcccg gtttggtggc cgtactttcg
     1081 tggtggggag tacgagccgg cactggaata tccgtggccg ctgatcagcg gccccccgag
     1141 gatcctgatc gcaaccgcaa acgctccgga gggtgaaaac ccgactctgg acgttgaggc
     1201 agagctgaag catctggaag aagcactggc agaaccgcgt aaacgtaaac tgtgtgaggt
     1261 acagcgttgc ccggcagcaa ccgtaagcga tatcaccagc gcactgcgta gctttaaacc
     1321 gcatatactg cattttatcg gtcatggtac tgcactcggt gtttatctga ggagcgcaga
     1381 acatgacggt gcacagtttg ttcgtggtga ggatttccag cagatgatcg caacctcgct
     1441 acgtcagaag gaccgtgaaa tgcatctggt tgttctgaac gcttgctgca cccacgaact
     1501 ggcaaaggca ctgaccgaac aggttagctg caccatcggt accgatatcg aagtttatga
     1561 ttcggcaagc atacattttg ctgcacgttt ttatgatcac ctggttcatg gtaccagcgt
     1621 tcattatgca tttaacgcag cagttgatga atgtcgtgca cacagcacca gcggccagga
     1681 agttttttgc ctgcacccgg cagcagaacg tgcaccggca agcgcaaccc cgccggtacg
     1741 tgcagatgaa ctggttttct ttagcccggc agaacgtcat agctaataac atacgcgtaa
     1801 caaggtaagg acaaattaat ggcagcaccg ggtagaccga ccccgaaagc actggatgaa
     1861 aaaagcctga agccgcatac tgttcacctg aaccgtgtta tccgtgaacg tctggttcaa
     1921 ctgcgtgaat cgctggcagc tagccgtgca ccgcgtctga cccaggatgc tcatccgcat
     1981 ctggttcgta tcgaagcagc acacgaattc cgtctcgcat atgaaaccag gatcaacaac
     2041 cgtcagcagg gggttctgac cgcagcatgg gcagacaccc tgtggagata ccgtgggctg
     2101 gcagttagct ggcaggcact ggcaagcgag atccaggcaa gggcacgtgc atttgttccc
     2161 atgcagtatg ttcacatcga tggcccgacc gaacgtcgtc tgttcagcct ggctcgtgag
     2221 ccggcacgtc atagcctggc aatcgttcag gaaagggacg gtaccctgtg gctggacggt
     2281 ggcagatttc agggtgttgc aagcggtgca cgtttcgcag ttctgccgcc gaccagcacc
     2341 gaaagcgaca tcgatagcgc actggcagaa gttcgtgcag ttgaagttat gccggaacgt
     2401 acccgtgttg aactggctac cgcagcagat gacatcccga gatggcaagg cctccatgca
     2461 tggccgctgc atgcacgtgg cgcttggacc ggtgtagcag ttcgtgctca gaccgcagca
     2521 gtttgtgctg cactcctgag agctatcgaa gcaagccagc gtatgcatct gtgtccggat
     2581 agcgacgggc cggttgctac cgttatcgaa ctatcgagcg atgagggcag caggatcgca
     2641 gttctggatg ggcacggtgt ttgtgttagc cccccggttg caacccccga acgtgcactc
     2701 gcactggttg aaaggctcca gagagctgca ctcgttcgta tgctggggag cggtaccgat
     2761 gagcatgcac taccgatccc gctcgcactc agactggaac gtgcagacgg tatcggtggt
     2821 acctgccgtc atctagcaga agggcgtacc gaaccgccga gcgaactgct actcgcaagc
     2881 gctggggcac aggtacatgc aatcgtagag aaccatggga gcgctatcgc aaccgatacc
     2941 caggatgttt atctgaccat gttctgtatc gcagcagacg gtcgtgttga aagagttagc
     3001 aagcagcaga gcagcgggat ccgtctgtgt gggggtgcaa agcataggct cgagtatgaa
     3061 ctaggtgcag ggggtgcaag cctgccggaa tacagggcag cagattcgag caccagcggc
     3121 tatgggctgc atagcctcat agttgttgca accgaccgtc agcaggagct gcgtaacctg
     3181 gaaacctggt cgggtgaagg tagcgctgca aaactgcagc cgggtgttct ggatctgacc
     3241 gctagcaaac gtgcaccggc tagacgtggc tcggctagcg caagcgggcc gctgagatat
     3301 gctgttctac gtgtagatct gcgtgtagct gcagcagtag cagggcgtga tgcaccgacc
     3361 ctgtaatgcc gtgggacagc ccgtcttaat taacctaggc tgctgccacc gctgagcaat
     3421 aactagcata accccttggg gcctctaaac gggtcttgag gggttttttg gggtcatcgt
     3481 ggccggatct tgcggcccct cggcttgaac gaattgttag acagattatc aaaaaggatc
     3541 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     3601 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     3661 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     3721 ggcttaccat ctggccccag tgctgcaatg ataccgcggg acccacgctc accggctcca
     3781 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     3841 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     3901 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     3961 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     4021 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     4081 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     4141 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     4201 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     4261 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     4321 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     4381 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     4441 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     4501 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     4561 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctgt agctagctca
     4621 ctcggtcgct acgctccggg cgtgagactg cggcgggcgc tgcggacaca tacaaagtta
     4681 cccacagatt ccgtggataa gcaggggact aacatgtgag gcaaaacagc agggccgcgc
     4741 cggtggcgtt tttccatagg ctccgccctc ctgccagagt tcacataaac agacgctttt
     4801 ccggtgcatc tgtgggagcc gtgaggctca accatgaatc tgacagtacg ggcgaaaccc
     4861 gacaggactt aaagatcccc accgtttccg gcgggtcgct ccctcttgcg ctctcctgtt
     4921 ccgaccctgc cgtttaccgg atacctgttc cgcctttctc ccttacggga agtgtggcgc
     4981 tttctcatag ctcacacact ggtatctcgg ctcggtgtag gtcgttcgct ccaagctggg
     5041 ctgtaagcaa gaactccccg ttcagcccga ctgctgcgcc ttatccggta actgttcact
     5101 tgagtccaac ccggaaaagc acggtaaaac gccactggca gcagccattg gtaactggga
     5161 gttcgcagag gatttgttta gctaaacacg cggttgctct tgaagtgtgc gccaaagtcc
     5221 ggctacactg gaaggacaga tttggttgct gtgctctgcg aaagccagtt accacggtta
     5281 agcagttccc caactgactt aaccttcgat caaaccacct ccccaggtgg ttttttcgtt
     5341 tacagggcaa aagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     5401 tactgaaccg ctctagattt cagtgcaatt tatctcttca aatgtagcac ctgaagtcag
     5461 ccccatacga tataagttgt aattctcatg ttagtcatgc cccgcgccca ccggaaggag
     5521 ctgactgggt tgaaggctct caagggcatc ggtcgagatc ccggtgccta atgagtgagc
     5581 taacttacat taattgcgtt gcgc
//
