LOCUS       sequence-419232-        3348 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  JAGBLH010000220 Cas8-HNH crRNA expression in pCOLADuet-1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3348)
  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 3348)
  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: pHS1929
FEATURES             Location/Qualifiers
     source          1..3348
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     18..37
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        18..36
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      125..172
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(402..1217)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(402..1217)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     1125..1144
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(1218..1309)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1327..1962)
                     /direction=LEFT
                     /label=ColA ori
                     /note="Plasmids containing the ColA origin of replication 
                     can be propagated in E. coli cells that contain additional 
                     plasmids with compatible origins."
     primer_bind     complement(1472..1491)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2160..3242)
                     /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     3204..3223
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3243..3320)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 ctcctgcatt aggaaattaa tacgactcac tatagggtgt accgccggat aggcggttta
       61 gaaggagaag tcatttaata aggccactgt taaaaagtgt accgccggat aggcggttta
      121 gaagctagca taaccccttg gggcctctaa acgggtcttg aggggttttt tgctgaaacc
      181 tcaggcattt gagaagcaca cggtcacact gcttccggta gtcaataaac cggtaaacca
      241 gcaatagaca taagcggcta tttaacgacc ctgccctgaa ccgacgacaa gctgacgacc
      301 gggtctccgc aagtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
      361 tctaaataca ttcaaatatg tatccgctca tgaattaatt cttagaaaaa ctcatcgagc
      421 atcaaatgaa actgcaattt attcatatca ggattatcaa taccatattt ttgaaaaagc
      481 cgtttctgta atgaaggaga aaactcaccg aggcagttcc ataggatggc aagatcctgg
      541 tatcggtctg cgattccgac tcgtccaaca tcaatacaac ctattaattt cccctcgtca
      601 aaaataaggt tatcaagtga gaaatcacca tgagtgacga ctgaatccgg tgagaatggc
      661 aaaagtttat gcatttcttt ccagacttgt tcaacaggcc agccattacg ctcgtcatca
      721 aaatcactcg catcaaccaa accgttattc attcgtgatt gcgcctgagc gagacgaaat
      781 acgcggtcgc tgttaaaagg acaattacaa acaggaatcg aatgcaaccg gcgcaggaac
      841 actgccagcg catcaacaat attttcacct gaatcaggat attcttctaa tacctggaat
      901 gctgttttcc cggggatcgc agtggtgagt aaccatgcat catcaggagt acggataaaa
      961 tgcttgatgg tcggaagagg cataaattcc gtcagccagt ttagtctgac catctcatct
     1021 gtaacatcat tggcaacgct acctttgcca tgtttcagaa acaactctgg cgcatcgggc
     1081 ttcccataca atcgatagat tgtcgcacct gattgcccga cattatcgcg agcccattta
     1141 tacccatata aatcagcatc catgttggaa tttaatcgcg gcctagagca agacgtttcc
     1201 cgttgaatat ggctcatact cttccttttt caatattatt gaagcattta tcagggttat
     1261 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggcatgcta
     1321 gcgcagaaac gtcctagaag atgccaggag gatacttagc agagagacaa taaggccgga
     1381 gcgaagccgt ttttccatag gctccgcccc cctgacgaac atcacgaaat ctgacgctca
     1441 aatcagtggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctgatggc
     1501 tccctcttgc gctctcctgt tcccgtcctg cggcgtccgt gttgtggtgg aggctttacc
     1561 caaatcacca cgtcccgttc cgtgtagaca gttcgctcca agctgggctg tgtgcaagaa
     1621 ccccccgttc agcccgactg ctgcgcctta tccggtaact atcatcttga gtccaacccg
     1681 gaaagacacg acaaaacgcc actggcagca gccattggta actgagaatt agtggattta
     1741 gatatcgaga gtcttgaagt ggtggcctaa cagaggctac actgaaagga cagtatttgg
     1801 tatctgcgct ccactaaagc cagttaccag gttaagcagt tccccaactg acttaacctt
     1861 cgatcaaacc gcctccccag gcggtttttt cgtttacaga gcaggagatt acgacgatcg
     1921 taaaaggatc tcaagaagat cctttacgga ttcccgacac catcactcta gatttcagtg
     1981 caatttatct cttcaaatgt agcacctgaa gtcagcccca tacgatataa gttgtaattc
     2041 tcatgttagt catgccccgc gcccaccgga aggagctgac tgggttgaag gctctcaagg
     2101 gcatcggtcg agatcccggt gcctaatgag tgagctaact tacattaatt gcgttgcgct
     2161 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     2221 gcgcggggag aggcggtttg cgtattgggc gccagggtgg tttttctttt caccagtgag
     2281 acgggcaaca gctgattgcc cttcaccgcc tggccctgag agagttgcag caagcggtcc
     2341 acgctggttt gccccagcag gcgaaaatcc tgtttgatgg tggttaacgg cgggatataa
     2401 catgagctgt cttcggtatc gtcgtatccc actaccgaga tgtccgcacc aacgcgcagc
     2461 ccggactcgg taatggcgcg cattgcgccc agcgccatct gatcgttggc aaccagcatc
     2521 gcagtgggaa cgatgccctc attcagcatt tgcatggttt gttgaaaacc ggacatggca
     2581 ctccagtcgc cttcccgttc cgctatcggc tgaatttgat tgcgagtgag atatttatgc
     2641 cagccagcca gacgcagacg cgccgagaca gaacttaatg ggcccgctaa cagcgcgatt
     2701 tgctggtgac ccaatgcgac cagatgctcc acgcccagtc gcgtaccgtc ttcatgggag
     2761 aaaataatac tgttgatggg tgtctggtca gagacatcaa gaaataacgc cggaacatta
     2821 gtgcaggcag cttccacagc aatggcatcc tggtcatcca gcggatagtt aatgatcagc
     2881 ccactgacgc gttgcgcgag aagattgtgc accgccgctt tacaggcttc gacgccgctt
     2941 cgttctacca tcgacaccac cacgctggca cccagttgat cggcgcgaga tttaatcgcc
     3001 gcgacaattt gcgacggcgc gtgcagggcc agactggagg tggcaacgcc aatcagcaac
     3061 gactgtttgc ccgccagttg ttgtgccacg cggttgggaa tgtaattcag ctccgccatc
     3121 gccgcttcca ctttttcccg cgttttcgca gaaacgtggc tggcctggtt caccacgcgg
     3181 gaaacggtct gataagagac accggcatac tctgcgacat cgtataacgt tactggtttc
     3241 acattcacca ccctgaattg actctcttcc gggcgctatc atgccatacc gcgaaaggtt
     3301 ttgcgccatt cgatggtgtc cgggatctcg acgctctccc ttatgcga
//
