LOCUS       sequence-430727-        4141 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression plasmid for SAVED-CHAT from Haliangium 
            ochraceum.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4141)
  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 4141)
  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
FEATURES             Location/Qualifiers
     source          1..4141
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(242..288)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     303..325
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     promoter        join(332..337,338..355,356..362)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     primer_bind     complement(2038..2056)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2052..2099
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(join(2364..3155,3156..3224))
                     /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     2987..3006
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3225..3329)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3403..4141)
                     /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 ttttctactg aaccgctcta gatttcagtg caatttatct cttcaaatgt agcacctgaa
       61 gtcagcccca tacgatataa gttgtaattc tcatgttagt catgccccgc gcccaccgga
      121 aggagctgac tgggttgaag gctctcaagg gcatcggtcg agatcccggt gcctaatgag
      181 tgagctaact tacattaatt gcgttgcgcc gactcctgca ttaggaaatg acaaacaaca
      241 gataaaacga aaggcccagt ctttcgactg agcctttcgt tttatttgat gcctttaatt
      301 aaagcggata acaatttcac acaggaggcc gtttacactt tatgcttccg gctcgtataa
      361 tgtgtgggga aaccacaacg gtttccctct agaaataatt ttgtttaact ataagaagga
      421 gatatacata tgactcacca tgctgatcat gactgccagc cgcacaggga gagcgttcag
      481 ccggatgaaa tccagtgtat cctggttctg gacctgagca tcgataacgc aatcaccgca
      541 tgtagcgtta ccccgcatct gccccgtgct gcacgtcgtg ttgaactaca tctgaacgac
      601 tttggtgcag aacgtgcacc gtacgggggt gcaagcgatc gtcgtacctg gcgttgttgg
      661 atgcaggcag ttgatgcaat gctggctgac gcacgtgcac agctgggtgc agaagttgaa
      721 tttactcatt attatctggc aggtcgtgca gcactcccgg tttttgcata tctgggtctg
      781 agactgggta aacaggcaaa catcactacc gttaaccgtc gtgatgacgg ttgttgggat
      841 gttgttccgt gccagcgtcc ggcttcgagc gcagcatcgg gggtttcgcc cagcgcacgt
      901 ttttttgacg aagttcgtgg tctggacacc gatgagcgta gcagcgaaag cggtatggtt
      961 gcagtttggg taagcaccca gcgtgatgtt gatcgtgggc tgctgcgtgc atttgcaagg
     1021 gcaagaggtg accgtgacct ggcaggtatc gttagcctgc gtgcacgtcc ggctgcaggt
     1081 gatgacaccg gtgatatgcg tctcctggaa ggtgcagacg gccccgatgc tgcacgtgaa
     1141 ctggttaact gttttcgtag catcccgaac cagtatccgc gtagcagcgg cctgatggtt
     1201 tttgttagcg gcccggttac cctggcagca atggttgggc gtgcaatcaa cccgagaatc
     1261 catggcccgg tttggtggcc gtactttcgt ggtggggagt acgagccggc actggaatat
     1321 ccgtggccgc tgatcagcgg ccccccgagg atcctgatcg caaccgcaaa cgctccggag
     1381 ggtgaaaacc cgactctgga cgttgaggca gagctgaagc atctggaaga agcactggca
     1441 gaaccgcgta aacgtaaact gtgtgaggta cagcgttgcc cggcagcaac cgtaagcgat
     1501 atcaccagcg cactgcgtag ctttaaaccg catatactgc attttatcgg tcatggtact
     1561 gcactcggtg tttatctgag gagcgcagaa catgacggtg cacagtttgt tcgtggtgag
     1621 gatttccagc agatgatcgc aacctcgcta cgtcagaagg accgtgaaat gcatctggtt
     1681 gttctgaacg cttgctgcac ccacgaactg gcaaaggcac tgaccgaaca ggttagctgc
     1741 accatcggta ccgatatcga agtttatgat tcggcaagca tacattttgc tgcacgtttt
     1801 tatgatcacc tggttcatgg taccagcgtt cattatgcat ttaacgcagc agttgatgaa
     1861 tgtcgtgcac acagcaccag cggccaggaa gttttttgcc tgcacccggc agcagaacgt
     1921 gcaccggcaa gcgcaacccc gccggtacgt gcagatgaac tggttttctt tagcccggca
     1981 gaacgtcata gctaatgccg tgggacagcc cgtcttaatt aacctaggct gctgccaccg
     2041 ctgagcaata actagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgc
     2101 tgaaacctca ggcatttgag aagcacacgg tcacactgct tccggtagtc aataaaccgg
     2161 taaaccagca atagacataa gcggctattt aacgaccctg ccctgaaccg acgaccgggt
     2221 catcgtggcc ggatcttgcg gcccctcggc ttgaacgaat tgttagacag attatcaaaa
     2281 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     2341 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     2401 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     2461 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgggaccc acgctcaccg
     2521 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     2581 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     2641 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     2701 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     2761 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     2821 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     2881 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     2941 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     3001 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     3061 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     3121 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     3181 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     3241 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     3301 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgtagct
     3361 agctcactcg gtcgctacgc tccgggcgtg agactgcggc gggcgctgcg gacacataca
     3421 aagttaccca cagattccgt ggataagcag gggactaaca tgtgaggcaa aacagcaggg
     3481 ccgcgccggt ggcgtttttc cataggctcc gccctcctgc cagagttcac ataaacagac
     3541 gcttttccgg tgcatctgtg ggagccgtga ggctcaacca tgaatctgac agtacgggcg
     3601 aaacccgaca ggacttaaag atccccaccg tttccggcgg gtcgctccct cttgcgctct
     3661 cctgttccga ccctgccgtt taccggatac ctgttccgcc tttctccctt acgggaagtg
     3721 tggcgctttc tcatagctca cacactggta tctcggctcg gtgtaggtcg ttcgctccaa
     3781 gctgggctgt aagcaagaac tccccgttca gcccgactgc tgcgccttat ccggtaactg
     3841 ttcacttgag tccaacccgg aaaagcacgg taaaacgcca ctggcagcag ccattggtaa
     3901 ctgggagttc gcagaggatt tgtttagcta aacacgcggt tgctcttgaa gtgtgcgcca
     3961 aagtccggct acactggaag gacagatttg gttgctgtgc tctgcgaaag ccagttacca
     4021 cggttaagca gttccccaac tgacttaacc ttcgatcaaa ccacctcccc aggtggtttt
     4081 ttcgtttaca gggcaaaaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     4141 c
//
