LOCUS       sequence-415370-        8263 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  NZ_CP025815 DinG HNH Cascade proteins operon (E. coli codon 
            optimized) with His14-tagged Csf3 in pET45b(+).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8263)
  AUTHORS   Altae-Tran H, Kannan S, Suberski AJ, Mears KS, Demircioglu FE, 
            Moeller L, Kocalar S, Oshiro R, Makarova KS, Macrae RK, Koonin EV, 
            Zhang F
  TITLE     Uncovering the functional diversity of rare CRISPR-Cas systems with 
            deep terascale clustering.
  JOURNAL   Science. 2023 Nov 24;382(6673):eadi1910. doi: 
            10.1126/science.adi1910. Epub 2023 Nov 23.
  PUBMED    37995242
REFERENCE   2  (bases 1 to 8263)
  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"; journalName: "Science."; 
            date: "2023-11-24"; volume: "382(6673):eadi1910. doi"; pages: " 
            10.1126/science.adi1910. Epub 2023 Nov 23"
COMMENT     Sequence Label: pHS1543
FEATURES             Location/Qualifiers
     source          1..8263
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     207..226
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     310..329
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        310..328
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    329..353
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     RBS             368..390
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     terminator      3087..3134
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(3309..3314,3315..3331,3332..3337))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(3364..3382)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3450..3554
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3555..3623,3624..4415)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3773..3792)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4586..5174
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5075..5094
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5328..5345
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5360..5499
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5425..5444)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5585..5607
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5601..5792)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(7104..8186)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     8148..8167
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(join(8187..8263,1))
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 ccgggatctc gacgctctcc cttatgcgac tcctgcatta ggaagcagcc cagtagtagg
       61 ttgaggccgt tgagcaccgc cgccgcaagg aatggtgcat gcaaggagat ggcgcccaac
      121 agtcccccgg ccacggggcc tgccaccata cccacgccga aacaagcgct catgagcccg
      181 aagtggcgag cccgatcttc cccatcggtg atgtcggcga tataggcgcc agcaaccgca
      241 cctgtggcgc cggtgatgcc ggccacgatg cgtccggcgt agaggatcga gatctcgatc
      301 ccgcgaaatt aatacgactc actatagggg aattgtgagc ggataacaat tcccctctag
      361 aaataatttt gtttaacttt aagaaggaga tataccatgg gcagcagcca tcaccatcat
      421 tcaggccatc accataccgg acaccaccat cattcaggca gtcatcacca ttccagcagc
      481 ggcgaaaatc tgatttttac ggcttcgctc aaatcacccc taatagctca aggatttatg
      541 acgtttgaca gcctgcttgc tgccgttctg ttcgatcgtt gtggtgacgt ggagactgcg
      601 catacctctg ttccgctgaa gtgctccgaa ggcctgttcc acgcgagcgc agcgattgtg
      661 accccggttg aacgtggtag ctacagcatt gcggcgtccc tgcgcgcgga ccacgatttg
      721 gacgcggact tattcccgaa aaacaaatct ggtcggattt atcgtaagat gggtcgtaag
      781 cgccgtcgtg atctcgggaa cgtgctgagc acctaccata cgatcagcac cccgtcggtg
      841 cgttggatct gcaccggcga cctggatcaa atcaaaggcc tgctgggcga cttgagattc
      901 atcggcaaga agcgcgcaca gggttttggt gaagttgccg cttggtcggt cgagaaaacc
      961 gatctggacc cgttggtcga tgaacagggc ctgccgcgtc gccctattcc ggagaacatg
     1021 tttaccggtg accgtagcct gccactgcaa gatctggcgt ggaaaccggc atattggaat
     1081 ccggagcacc gcgcagcttg ttacgcgcct cagccgcttt gaaggagggc cggatgaata
     1141 cctttgaact catatccaag ggctttgcta ggcacgtact aggaaattca acaacgggcg
     1201 acccgattcg tgatgctgcc aaggcgctga cgatgaacca cctgctgaac aacgagaaag
     1261 gcacttctag ccgtttgacc ggttgggcga ttttccgcgc tagcggtagc accgtccagg
     1321 cgaacttggc ggctaagggt gtgatggcga acgacggccg tacccgttac gaggcgttcg
     1381 cggatgagct gtcggatttc agagaaccgg tcgtgttcct gcaatttacc aagtccaccg
     1441 tgaatttagg taatgttttt gcgacctttg acccacgtgt taccaaaatc tgcgcaccgg
     1501 atgttgcacc ggagatcttt gacttcagcg ttggtacgct gccggaaacc gcaggcgtgg
     1561 ccgaacgtaa tatccgcaaa gccgttctga agaaggagaa aagcaatgct gcatgatcac
     1621 gtcatcccca agccgcccac actaggagca tatgctagtc accaattgct ggagggcctg
     1681 actgacggcg caagaccact gtttgttgat aatggcgacc acctcttggt tcgttccgtg
     1741 gatcaatcgc tgacccaatc tagcaaaccg gtcccgacgg tgaacctggg tgacgtgtta
     1801 ggcttcgaac tgcgcgccag ctgcggtgtg aaggtgaagg gtaaacatcg ttacttcccg
     1861 ctgaaggact ggcgtgctcg ccatgcgtgg ctgaatcgtc gtgcgagcgg ttttgatgtc
     1921 ctggctgtga ccgttagcgc acgctccacc ccgattaaaa agaacatccg tattgatcag
     1981 agcgattttg cgggtgtatt gcgtgttacc gacatcgaag cgttcaacca ggttcttgag
     2041 cacggcatcg gcggtccggg taaagccttc ggccacggaa tgctggtaat ctgaaggaga
     2101 acaccatgaa ccaatcaact ttaattaccg aagtcgtaat aaggttagaa gctcccctat
     2161 caattgctat gccggtggca gagtacacca agccgaacaa atgggaaaac tttccggtgg
     2221 ctccggtggg ttatgatgaa gagggtcaac cgatccagtc aggcttcctg ccggcaacca
     2281 ccattcgtgg tgcgatccgc cgttttgcgg tgctgccaga aatggcagcg gctgcggaga
     2341 agggtgcgca ctaccgcctg gagcgtgcat acagcgaact cattggtcag gacccgggct
     2401 ccgaacagca gccggacgag atcgatctga ctgccatcgc cgaagcgcgt gaggaatccc
     2461 cgattgttga tctgtttggc tctggcctgg gtgtcaaaag ccgcttgctg gtgagcaatt
     2521 tcgtgccgga ccatccgacg ctgccgatgg cgttcacctg ggttcgtaag gacatcggcg
     2581 acacggaagg tgttttggag gcgttgccag acgaggatcg ttctgcgtac ctggcgcgtg
     2641 agcaagcgaa ccgtcaccgt gcccaagcgg aggccttggt tgcggacctg aagcgcaaag
     2701 aatcgaaagc cgttaaaacc ggttccgatg atgcagctga cttgaaggtg cagctggaag
     2761 ctgctgagaa gctcgcggct aaatataaag aggacatggg tgacatgcaa gtttccaccc
     2821 gtagcctggg agttagctat gcgttaccgc agggtctgga gctcaagggg aagttcgtcg
     2881 tgcgtagcgc gcgcgagagg gatcaggaca tcctgctgcg cggcctggat ggcttatctc
     2941 gttggcccgt attgggcgca caaagcgcga gaggcgcagg tgaaattagc ggtactgcga
     3001 gctttaaact ggacggcaaa attaccaaga ttgtttcgtt cggcggtttc gaaccgctaa
     3061 aaattcgcga ctttgaagca acttaactag cataacccct tggggcctct aaacgggtct
     3121 tgaggggttt tttgctgaaa ggaggaacta tatccggata tcccgcaaga ggcccggcag
     3181 taccggcata accaagccta tgcctacagc atccagggtg acggtgccga ggatgacgat
     3241 gagcgcattg ttagatttca tacacggtgc ctgactgcgt tagcaattta actgtgataa
     3301 actaccgcat taaagcttat cgatgataag ctgtcaaaca tgagaattct tgaagacgaa
     3361 agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga
     3421 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     3481 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     3541 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     3601 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     3661 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     3721 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     3781 gcgcggtatt atcccgtgtt gacgccgggc aagagcaact cggtcgccgc atacactatt
     3841 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     3901 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     3961 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     4021 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     4081 gtgacaccac gatgcctgca gcaatggcaa caacgttgcg caaactatta actggcgaac
     4141 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     4201 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     4261 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     4321 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     4381 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     4441 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     4501 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     4561 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     4621 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     4681 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag
     4741 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     4801 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     4861 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     4921 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     4981 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     5041 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     5101 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     5161 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     5221 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     5281 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     5341 gcgaggaagc ggaagagcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt
     5401 cacaccgcaa tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta
     5461 tacactccgc tatcgctacg tgactgggtc atggctgcgc cccgacaccc gccaacaccc
     5521 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc
     5581 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag
     5641 ctgcggtaaa gctcatcagc gtggtcgtga agcgattcac agatgtctgc ctgttcatcc
     5701 gcgtccagct cgttgagttt ctccagaagc gttaatgtct ggcttctgat aaagcgggcc
     5761 atgttaaggg cggttttttc ctgtttggtc actgatgcct ccgtgtaagg gggatttctg
     5821 ttcatggggg taatgatacc gatgaaacga gagaggatgc tcacgatacg ggttactgat
     5881 gatgaacatg cccggttact ggaacgttgt gagggtaaac aactggcggt atggatgcgg
     5941 cgggaccaga gaaaaatcac tcagggtcaa tgccagcgct tcgttaatac agatgtaggt
     6001 gttccacagg gtagccagca gcatcctgcg atgcagatcc ggaacataat ggtgcagggc
     6061 gctgacttcc gcgtttacag actttacgaa acacggaaac cgaagaccat tcatgttgtt
     6121 gctcaggtcg cagacgtttt gcagcagcag tcgcttcacg ttcgctcgcg tatcggtgat
     6181 tcattctgct aaccagtaag gcaaccccgc cagcctagcc gggtcctcaa cgacaggagc
     6241 acgatcatgc gcacccgtgg ccaggaccca acgctgcccg agatgcgccg cgtgcggctg
     6301 ctggagatgg cggacgcgat ggatatgttc tgccaagggt tggtttgcgc attcacagtt
     6361 ctccgcaaga attgattggc tccaattctt ggagtggtga atccgttagc gaggtgccgc
     6421 cggcttccat tcaggtcgag gtggcccggc tccatgcacc gcgacgcaac gcggggaggc
     6481 agacaaggta tagggcggcg cctacaatcc atgccaaccc gttccatgtg ctcgccgagg
     6541 cggcataaat cgccgtgacg atcagcggtc caatgatcga agttaggctg gtaagagccg
     6601 cgagcgatcc ttgaagctgt ccctgatggt cgtcatctac ctgcctggac agcatggcct
     6661 gcaacgcggg catcccgatg ccgccggaag cgagaagaat cataatgggg aaggccatcc
     6721 agcctcgcgt cgcgaacgcc agcaagacgt agcccagcgc gtcggccgcc atgccggcga
     6781 taatggcctg cttctcgccg aaacgtttgg tggcgggacc agtgacgaag gcttgagcga
     6841 gggcgtgcaa gattccgaat accgcaagcg acaggccgat catcgtcgcg ctccagcgaa
     6901 agcggtcctc gccgaaaatg acccagagcg ctgccggcac ctgtcctacg agttgcatga
     6961 taaagaagac agtcataagt gcggcgacga tagtcatgcc ccgcgcccac cggaaggagc
     7021 tgactgggtt gaaggctctc aagggcatcg gtcgagatcc cggtgcctaa tgagtgagct
     7081 aacttacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     7141 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgccagg
     7201 gtggtttttc ttttcaccag tgagacgggc aacagctgat tgcccttcac cgcctggccc
     7261 tgagagagtt gcagcaagcg gtccacgctg gtttgcccca gcaggcgaaa atcctgtttg
     7321 atggtggtta acggcgggat ataacatgag ctgtcttcgg tatcgtcgta tcccactacc
     7381 gagatatccg caccaacgcg cagcccggac tcggtaatgg cgcgcattgc gcccagcgcc
     7441 atctgatcgt tggcaaccag catcgcagtg ggaacgatgc cctcattcag catttgcatg
     7501 gtttgttgaa aaccggacat ggcactccag tcgccttccc gttccgctat cggctgaatt
     7561 tgattgcgag tgagatattt atgccagcca gccagacgca gacgcgccga gacagaactt
     7621 aatgggcccg ctaacagcgc gatttgctgg tgacccaatg cgaccagatg ctccacgccc
     7681 agtcgcgtac cgtcttcatg ggagaaaata atactgttga tgggtgtctg gtcagagaca
     7741 tcaagaaata acgccggaac attagtgcag gcagcttcca cagcaatggc atcctggtca
     7801 tccagcggat agttaatgat cagcccactg acgcgttgcg cgagaagatt gtgcaccgcc
     7861 gctttacagg cttcgacgcc gcttcgttct accatcgaca ccaccacgct ggcacccagt
     7921 tgatcggcgc gagatttaat cgccgcgaca atttgcgacg gcgcgtgcag ggccagactg
     7981 gaggtggcaa cgccaatcag caacgactgt ttgcccgcca gttgttgtgc cacgcggttg
     8041 ggaatgtaat tcagctccgc catcgccgct tccacttttt cccgcgtttt cgcagaaacg
     8101 tggctggcct ggttcaccac gcgggaaacg gtctgataag agacaccggc atactctgcg
     8161 acatcgtata acgttactgg tttcacattc accaccctga attgactctc ttccgggcgc
     8221 tatcatgcca taccgcgaaa ggttttgcgc cattcgatgg tgt
//
