LOCUS       sequence-426381-        7205 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression plasmid for SAVED-CHAT, PCaspase, PCi, and 
            PC-sigma from Haliangium ochraceum.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7205)
  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 7205)
  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_PCi_PC-sigma
FEATURES             Location/Qualifiers
     source          1..7205
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(1..792,793..861))
                     /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     624..643
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(862..966)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1040..1778)
                     /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."
     terminator      complement(2020..2066)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     2081..2103
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     promoter        join(2110..2115,2116..2133,2134..2140)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     regulatory      4039..4048
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(6997..7015)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      7011..7058
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
       61 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
      121 cagtgctgca atgataccgc gggacccacg ctcaccggct ccagatttat cagcaataaa
      181 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
      241 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
      301 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
      361 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
      421 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
      481 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
      541 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
      601 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
      661 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
      721 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
      781 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
      841 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
      901 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
      961 tccgcgcaca tttccccgaa aagtgccacc tgtagctagc tcactcggtc gctacgctcc
     1021 gggcgtgaga ctgcggcggg cgctgcggac acatacaaag ttacccacag attccgtgga
     1081 taagcagggg actaacatgt gaggcaaaac agcagggccg cgccggtggc gtttttccat
     1141 aggctccgcc ctcctgccag agttcacata aacagacgct tttccggtgc atctgtggga
     1201 gccgtgaggc tcaaccatga atctgacagt acgggcgaaa cccgacagga cttaaagatc
     1261 cccaccgttt ccggcgggtc gctccctctt gcgctctcct gttccgaccc tgccgtttac
     1321 cggatacctg ttccgccttt ctcccttacg ggaagtgtgg cgctttctca tagctcacac
     1381 actggtatct cggctcggtg taggtcgttc gctccaagct gggctgtaag caagaactcc
     1441 ccgttcagcc cgactgctgc gccttatccg gtaactgttc acttgagtcc aacccggaaa
     1501 agcacggtaa aacgccactg gcagcagcca ttggtaactg ggagttcgca gaggatttgt
     1561 ttagctaaac acgcggttgc tcttgaagtg tgcgccaaag tccggctaca ctggaaggac
     1621 agatttggtt gctgtgctct gcgaaagcca gttaccacgg ttaagcagtt ccccaactga
     1681 cttaaccttc gatcaaacca cctccccagg tggttttttc gtttacaggg caaaagatta
     1741 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctactgaa ccgctctaga
     1801 tttcagtgca atttatctct tcaaatgtag cacctgaagt cagccccata cgatataagt
     1861 tgtaattctc atgttagtca tgccccgcgc ccaccggaag gagctgactg ggttgaaggc
     1921 tctcaagggc atcggtcgag atcccggtgc ctaatgagtg agctaactta cattaattgc
     1981 gttgcgccga ctcctgcatt aggaaatgac aaacaacaga taaaacgaaa ggcccagtct
     2041 ttcgactgag cctttcgttt tatttgatgc ctttaattaa agcggataac aatttcacac
     2101 aggaggccgt ttacacttta tgcttccggc tcgtataatg tgtggggaaa ccacaacggt
     2161 ttccctctag aaataatttt gtttaactat aagaaggaga tatacatatg actcaccatg
     2221 ctgatcatga ctgccagccg cacagggaga gcgttcagcc ggatgaaatc cagtgtatcc
     2281 tggttctgga cctgagcatc gataacgcaa tcaccgcatg tagcgttacc ccgcatctgc
     2341 cccgtgctgc acgtcgtgtt gaactacatc tgaacgactt tggtgcagaa cgtgcaccgt
     2401 acgggggtgc aagcgatcgt cgtacctggc gttgttggat gcaggcagtt gatgcaatgc
     2461 tggctgacgc acgtgcacag ctgggtgcag aagttgaatt tactcattat tatctggcag
     2521 gtcgtgcagc actcccggtt tttgcatatc tgggtctgag actgggtaaa caggcaaaca
     2581 tcactaccgt taaccgtcgt gatgacggtt gttgggatgt tgttccgtgc cagcgtccgg
     2641 cttcgagcgc agcatcgggg gtttcgccca gcgcacgttt ttttgacgaa gttcgtggtc
     2701 tggacaccga tgagcgtagc agcgaaagcg gtatggttgc agtttgggta agcacccagc
     2761 gtgatgttga tcgtgggctg ctgcgtgcat ttgcaagggc aagaggtgac cgtgacctgg
     2821 caggtatcgt tagcctgcgt gcacgtccgg ctgcaggtga tgacaccggt gatatgcgtc
     2881 tcctggaagg tgcagacggc cccgatgctg cacgtgaact ggttaactgt tttcgtagca
     2941 tcccgaacca gtatccgcgt agcagcggcc tgatggtttt tgttagcggc ccggttaccc
     3001 tggcagcaat ggttgggcgt gcaatcaacc cgagaatcca tggcccggtt tggtggccgt
     3061 actttcgtgg tggggagtac gagccggcac tggaatatcc gtggccgctg atcagcggcc
     3121 ccccgaggat cctgatcgca accgcaaacg ctccggaggg tgaaaacccg actctggacg
     3181 ttgaggcaga gctgaagcat ctggaagaag cactggcaga accgcgtaaa cgtaaactgt
     3241 gtgaggtaca gcgttgcccg gcagcaaccg taagcgatat caccagcgca ctgcgtagct
     3301 ttaaaccgca tatactgcat tttatcggtc atggtactgc actcggtgtt tatctgagga
     3361 gcgcagaaca tgacggtgca cagtttgttc gtggtgagga tttccagcag atgatcgcaa
     3421 cctcgctacg tcagaaggac cgtgaaatgc atctggttgt tctgaacgct tgctgcaccc
     3481 acgaactggc aaaggcactg accgaacagg ttagctgcac catcggtacc gatatcgaag
     3541 tttatgattc ggcaagcata cattttgctg cacgttttta tgatcacctg gttcatggta
     3601 ccagcgttca ttatgcattt aacgcagcag ttgatgaatg tcgtgcacac agcaccagcg
     3661 gccaggaagt tttttgcctg cacccggcag cagaacgtgc accggcaagc gcaaccccgc
     3721 cggtacgtgc agatgaactg gttttcttta gcccggcaga acgtcatagc taataacata
     3781 cgcgtaacaa ggtaaggaca aattaatgat gaaaagggct ctgctgatag gtgcaccgct
     3841 ggcacatagc gcaacccatg cactggcagg tgttgcaaac gatatcgacc gtgcaggtga
     3901 aatcctgcgt ctgtatggtt ttcattgtga tgacgttctc tttggtgaac gtgcaactag
     3961 agatggtatc gttgcacgtc tggaagcact gatcgctgaa acccaggctg atgatgctgt
     4021 tgttatctat tttagcggcc atggtggccg tgttgttaac accgggatcg ttaaagatct
     4081 accgggtggt gactatcgtc ccggcgctca ccagttcctg gttccggaag actaccaacc
     4141 gaaagcacag acctttaccg gtatcctgga cttcgagctg aggtatctag ttgcacgtct
     4201 ggcagcacgt accgaaaacg ttagcgttat cctggattgt tgtcatagcg gtggtgcaat
     4261 aagagcaccg ggtagaccga ccccgaaagc actggatgaa aaaagcctga agccgcatac
     4321 tgttcacctg aaccgtgtta tccgtgaacg tctggttcaa ctgcgtgaat cgctggcagc
     4381 tagccgtgca ccgcgtctga cccaggatgc tcatccgcat ctggttcgta tcgaagcagc
     4441 acacgaattc cgtctcgcat atgaaaccag gatcaacaac cgtcagcagg gggttctgac
     4501 cgcagcatgg gcagacaccc tgtggagata ccgtgggctg gcagttagct ggcaggcact
     4561 ggcaagcgag atccaggcaa gggcacgtgc atttgttccc atgcagtatg ttcacatcga
     4621 tggcccgacc gaacgtcgtc tgttcagcct ggctcgtgag ccggcacgtc atagcctggc
     4681 aatcgttcag gaaagggacg gtaccctgtg gctggacggt ggcagatttc agggtgttgc
     4741 aagcggtgca cgtttcgcag ttctgccgcc gaccagcacc gaaagcgaca tcgatagcgc
     4801 actggcagaa gttcgtgcag ttgaagttat gccggaacgt acccgtgttg aactggctac
     4861 cgcagcagat gacatcccga gatggcaagg cctccatgca tggccgctgc atgcacgtgg
     4921 cgcttggacc ggtgtagcag ttcgtgctca gaccgcagca gtttgtgctg cactcctgag
     4981 agctatcgaa gcaagccagc gtatgcatct gtgtccggat agcgacgggc cggttgctac
     5041 cgttatcgaa ctatcgagcg atgagggcag caggatcgca gttctggatg ggcacggtgt
     5101 ttgtgttagc cccccggttg caacccccga acgtgcactc gcactggttg aaaggctcca
     5161 gagagctgca ctcgttcgta tgctggggag cggtaccgat gagcatgcac taccgatccc
     5221 gctcgcactc agactggaac gtgcagacgg tatcggtggt acctgccgtc atctagcaga
     5281 agggcgtacc gaaccgccga gcgaactgct actcgcaagc gctggggcac aggtacatgc
     5341 aatcgtagag aaccatggga gcgctatcgc aaccgatacc caggatgttt atctgaccat
     5401 gttctgtatc gcagcagacg gtcgtgttga aagagttagc aagcagcaga gcagcgggat
     5461 ccgtctgtgt gggggtgcaa agcataggct cgagtatgaa ctaggtgcag ggggtgcaag
     5521 cctgccggaa tacagggcag cagattcgag caccagcggc tatgggctgc atagcctcat
     5581 agttgttgca accgaccgtc agcaggagct gcgtaacctg gaaacctggt cgggtgaagg
     5641 tagcgctgca aaactgcagc cgggtgttct ggatctgacc gctagcaaac gtgcaccggc
     5701 tagacgtggc tcggctagcg caagcgggcc gctgagatat gctgttctac gtgtagatct
     5761 gcgtgtagct gcagcagtag cagggcgtga tgcaccgacc ctgtaaatca catgaataag
     5821 gaggatgtcg tatgcacagg attgacgttt cgggtccgct gtacctggat tgtcaacaga
     5881 gcggggacac cgttcagctg attcaccaaa ccaccgttct ggttgaaccg gcaattgaac
     5941 tatacgggac cgataccagc accaccctgg aaattcgtga ctttgaagat gaaaacggaa
     6001 gatgtcgtgt taacgttttt accattgaac gtgacggtac cctgaccccg tgggaacgtc
     6061 tagcaagcgg agcagtttcg ccggcagttt ggagcattat tgagcgtgga ggggagcgtg
     6121 ctgctctgct ggttccggtt ccgtacgcac cgctgggtga actgaccgca gacctggttg
     6181 atcaatttgc accgcaggat tacaccccgt cgagggttag aattgttctc ccgccgccgg
     6241 aagacgataa accgccgccg cacgatgacc aataaggaaa ggaggtaagg agggtcgcaa
     6301 atggatcatg atcaccgtga tgatgatgaa ggtaaacccg cactggctga agatagcgaa
     6361 tacattgatg agaacgttct aggtgacgca gatgttctgg aagatgatga tgaacgtggg
     6421 gaaagcgatg gtgatgaagt tagcgacgct gaactctttg caagctgggc tgcaggtgat
     6481 aagcatgcag cagatgttct gattcgtcgt tacatggata aactgagcgg ttactttgct
     6541 gcaaaactgt cgcagggtga tagcgttgca gacctggttc agcagaccct ggagggttgc
     6601 catcgtgcag caccgagctt ccgtggtgaa tgctcgtttc gtacctttct gtacagggtt
     6661 gcagggaaca aactgcgtga ctttctgcgt ggtaaggcac gtaggccccg tgcagaagac
     6721 attgatgaaa taagcgtagc agatctgaac ccgggaccga gcaccattcg tcgtcaaaac
     6781 caccgtcagg cagttctgct ggacgcactg cgtagcctgc cgctgcagat gcaggagctg
     6841 ctggaattcc gttaccagca agacctgagc atacgtgaaa ttgcagaaat tctgggtatt
     6901 gaggttagcg caaccaaaag ccgtctgcgt cgtgcaaagg aagcactgcg tagggcactg
     6961 ggaactagcg gtgctgatgc actgtagctg ctgccaccgc tgagcaataa ctagcataac
     7021 cccttggggc ctctaaacgg gtcttgaggg gttttttggg gtcatcgtgg ccggatcttg
     7081 cggcccctcg gcttgaacga attgttagac agattatcaa aaaggatctt cacctagatc
     7141 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     7201 gacag
//
