LOCUS       sequence-426426-        6086 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression plasmid for SAVED-CHAT, PCaspase N-terminal 
            fragment (aa 1-153), and C-terminal fragment (154-666) from 
            Haliangium ochraceum.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6086)
  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 6086)
  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-N-C-term
FEATURES             Location/Qualifiers
     source          1..6086
                     /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"
     regulatory      2052..2061
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(3891..3909)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3905..3952
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(join(4100..4891,4892..4960))
                     /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     4723..4742
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4961..5065)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5139..5877)
                     /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 gatgaaaagg gctctgctga taggtgcacc gctggcacat
     1861 agcgcaaccc atgcactggc aggtgttgca aacgatatcg accgtgcagg tgaaatcctg
     1921 cgtctgtatg gttttcattg tgatgacgtt ctctttggtg aacgtgcaac tagagatggt
     1981 atcgttgcac gtctggaagc actgatcgct gaaacccagg ctgatgatgc tgttgttatc
     2041 tattttagcg gccatggtgg ccgtgttgtt aacaccggga tcgttaaaga tctaccgggt
     2101 ggtgactatc gtcccggcgc tcaccagttc ctggttccgg aagactacca accgaaagca
     2161 cagaccttta ccggtatcct ggacttcgag ctgaggtatc tagttgcacg tctggcagca
     2221 cgtaccgaaa acgttagcgt tatcctggat tgttgtcata gcggtggtgc aataagataa
     2281 aaattttgca ccaattaggt tgtatggcac cgggtagacc gaccccgaaa gcactggatg
     2341 aaaaaagcct gaagccgcat actgttcacc tgaaccgtgt tatccgtgaa cgtctggttc
     2401 aactgcgtga atcgctggca gctagccgtg caccgcgtct gacccaggat gctcatccgc
     2461 atctggttcg tatcgaagca gcacacgaat tccgtctcgc atatgaaacc aggatcaaca
     2521 accgtcagca gggggttctg accgcagcat gggcagacac cctgtggaga taccgtgggc
     2581 tggcagttag ctggcaggca ctggcaagcg agatccaggc aagggcacgt gcatttgttc
     2641 ccatgcagta tgttcacatc gatggcccga ccgaacgtcg tctgttcagc ctggctcgtg
     2701 agccggcacg tcatagcctg gcaatcgttc aggaaaggga cggtaccctg tggctggacg
     2761 gtggcagatt tcagggtgtt gcaagcggtg cacgtttcgc agttctgccg ccgaccagca
     2821 ccgaaagcga catcgatagc gcactggcag aagttcgtgc agttgaagtt atgccggaac
     2881 gtacccgtgt tgaactggct accgcagcag atgacatccc gagatggcaa ggcctccatg
     2941 catggccgct gcatgcacgt ggcgcttgga ccggtgtagc agttcgtgct cagaccgcag
     3001 cagtttgtgc tgcactcctg agagctatcg aagcaagcca gcgtatgcat ctgtgtccgg
     3061 atagcgacgg gccggttgct accgttatcg aactatcgag cgatgagggc agcaggatcg
     3121 cagttctgga tgggcacggt gtttgtgtta gccccccggt tgcaaccccc gaacgtgcac
     3181 tcgcactggt tgaaaggctc cagagagctg cactcgttcg tatgctgggg agcggtaccg
     3241 atgagcatgc actaccgatc ccgctcgcac tcagactgga acgtgcagac ggtatcggtg
     3301 gtacctgccg tcatctagca gaagggcgta ccgaaccgcc gagcgaactg ctactcgcaa
     3361 gcgctggggc acaggtacat gcaatcgtag agaaccatgg gagcgctatc gcaaccgata
     3421 cccaggatgt ttatctgacc atgttctgta tcgcagcaga cggtcgtgtt gaaagagtta
     3481 gcaagcagca gagcagcggg atccgtctgt gtgggggtgc aaagcatagg ctcgagtatg
     3541 aactaggtgc agggggtgca agcctgccgg aatacagggc agcagattcg agcaccagcg
     3601 gctatgggct gcatagcctc atagttgttg caaccgaccg tcagcaggag ctgcgtaacc
     3661 tggaaacctg gtcgggtgaa ggtagcgctg caaaactgca gccgggtgtt ctggatctga
     3721 ccgctagcaa acgtgcaccg gctagacgtg gctcggctag cgcaagcggg ccgctgagat
     3781 atgctgttct acgtgtagat ctgcgtgtag ctgcagcagt agcagggcgt gatgcaccga
     3841 ccctgtaatg ccgtgggaca gcccgtctta attaacctag gctgctgcca ccgctgagca
     3901 ataactagca taaccccttg gggcctctaa acgggtcttg aggggttttt tggggtcatc
     3961 gtggccggat cttgcggccc ctcggcttga acgaattgtt agacagatta tcaaaaagga
     4021 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     4081 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     4141 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     4201 agggcttacc atctggcccc agtgctgcaa tgataccgcg ggacccacgc tcaccggctc
     4261 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     4321 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     4381 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     4441 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     4501 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     4561 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     4621 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     4681 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     4741 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     4801 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     4861 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     4921 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     4981 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     5041 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gtagctagct
     5101 cactcggtcg ctacgctccg ggcgtgagac tgcggcgggc gctgcggaca catacaaagt
     5161 tacccacaga ttccgtggat aagcagggga ctaacatgtg aggcaaaaca gcagggccgc
     5221 gccggtggcg tttttccata ggctccgccc tcctgccaga gttcacataa acagacgctt
     5281 ttccggtgca tctgtgggag ccgtgaggct caaccatgaa tctgacagta cgggcgaaac
     5341 ccgacaggac ttaaagatcc ccaccgtttc cggcgggtcg ctccctcttg cgctctcctg
     5401 ttccgaccct gccgtttacc ggatacctgt tccgcctttc tcccttacgg gaagtgtggc
     5461 gctttctcat agctcacaca ctggtatctc ggctcggtgt aggtcgttcg ctccaagctg
     5521 ggctgtaagc aagaactccc cgttcagccc gactgctgcg ccttatccgg taactgttca
     5581 cttgagtcca acccggaaaa gcacggtaaa acgccactgg cagcagccat tggtaactgg
     5641 gagttcgcag aggatttgtt tagctaaaca cgcggttgct cttgaagtgt gcgccaaagt
     5701 ccggctacac tggaaggaca gatttggttg ctgtgctctg cgaaagccag ttaccacggt
     5761 taagcagttc cccaactgac ttaaccttcg atcaaaccac ctccccaggt ggttttttcg
     5821 tttacagggc aaaagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     5881 tctactgaac cgctctagat ttcagtgcaa tttatctctt caaatgtagc acctgaagtc
     5941 agccccatac gatataagtt gtaattctca tgttagtcat gccccgcgcc caccggaagg
     6001 agctgactgg gttgaaggct ctcaagggca tcggtcgaga tcccggtgcc taatgagtga
     6061 gctaacttac attaattgcg ttgcgc
//
