LOCUS       sequence-416245-        3350 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MWEE01000013 Cas5-HNH crRNA expression in pCOLADuet-1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3350)
  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 3350)
  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: pHS1398
FEATURES             Location/Qualifiers
     source          1..3350
                     /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      127..174
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(404..1219)
                     /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(404..1219)
                     /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     1127..1146
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(1220..1311)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1329..1964)
                     /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(1474..1493)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2162..3244)
                     /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     3206..3225
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3245..3322)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 ctcctgcatt aggaaattaa tacgactcac tatagggtat tccccacgca tgtgggggtg
       61 aaccggagaa gtcatttaat aaggccactg ttaaaaagta ttccccacgc atgtgggggt
      121 gaaccgctag cataacccct tggggcctct aaacgggtct tgaggggttt tttgctgaaa
      181 cctcaggcat ttgagaagca cacggtcaca ctgcttccgg tagtcaataa accggtaaac
      241 cagcaataga cataagcggc tatttaacga ccctgccctg aaccgacgac aagctgacga
      301 ccgggtctcc gcaagtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
      361 tttctaaata cattcaaata tgtatccgct catgaattaa ttcttagaaa aactcatcga
      421 gcatcaaatg aaactgcaat ttattcatat caggattatc aataccatat ttttgaaaaa
      481 gccgtttctg taatgaagga gaaaactcac cgaggcagtt ccataggatg gcaagatcct
      541 ggtatcggtc tgcgattccg actcgtccaa catcaataca acctattaat ttcccctcgt
      601 caaaaataag gttatcaagt gagaaatcac catgagtgac gactgaatcc ggtgagaatg
      661 gcaaaagttt atgcatttct ttccagactt gttcaacagg ccagccatta cgctcgtcat
      721 caaaatcact cgcatcaacc aaaccgttat tcattcgtga ttgcgcctga gcgagacgaa
      781 atacgcggtc gctgttaaaa ggacaattac aaacaggaat cgaatgcaac cggcgcagga
      841 acactgccag cgcatcaaca atattttcac ctgaatcagg atattcttct aatacctgga
      901 atgctgtttt cccggggatc gcagtggtga gtaaccatgc atcatcagga gtacggataa
      961 aatgcttgat ggtcggaaga ggcataaatt ccgtcagcca gtttagtctg accatctcat
     1021 ctgtaacatc attggcaacg ctacctttgc catgtttcag aaacaactct ggcgcatcgg
     1081 gcttcccata caatcgatag attgtcgcac ctgattgccc gacattatcg cgagcccatt
     1141 tatacccata taaatcagca tccatgttgg aatttaatcg cggcctagag caagacgttt
     1201 cccgttgaat atggctcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     1261 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggcatgc
     1321 tagcgcagaa acgtcctaga agatgccagg aggatactta gcagagagac aataaggccg
     1381 gagcgaagcc gtttttccat aggctccgcc cccctgacga acatcacgaa atctgacgct
     1441 caaatcagtg gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctgatg
     1501 gctccctctt gcgctctcct gttcccgtcc tgcggcgtcc gtgttgtggt ggaggcttta
     1561 cccaaatcac cacgtcccgt tccgtgtaga cagttcgctc caagctgggc tgtgtgcaag
     1621 aaccccccgt tcagcccgac tgctgcgcct tatccggtaa ctatcatctt gagtccaacc
     1681 cggaaagaca cgacaaaacg ccactggcag cagccattgg taactgagaa ttagtggatt
     1741 tagatatcga gagtcttgaa gtggtggcct aacagaggct acactgaaag gacagtattt
     1801 ggtatctgcg ctccactaaa gccagttacc aggttaagca gttccccaac tgacttaacc
     1861 ttcgatcaaa ccgcctcccc aggcggtttt ttcgtttaca gagcaggaga ttacgacgat
     1921 cgtaaaagga tctcaagaag atcctttacg gattcccgac accatcactc tagatttcag
     1981 tgcaatttat ctcttcaaat gtagcacctg aagtcagccc catacgatat aagttgtaat
     2041 tctcatgtta gtcatgcccc gcgcccaccg gaaggagctg actgggttga aggctctcaa
     2101 gggcatcggt cgagatcccg gtgcctaatg agtgagctaa cttacattaa ttgcgttgcg
     2161 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     2221 acgcgcgggg agaggcggtt tgcgtattgg gcgccagggt ggtttttctt ttcaccagtg
     2281 agacgggcaa cagctgattg cccttcaccg cctggccctg agagagttgc agcaagcggt
     2341 ccacgctggt ttgccccagc aggcgaaaat cctgtttgat ggtggttaac ggcgggatat
     2401 aacatgagct gtcttcggta tcgtcgtatc ccactaccga gatgtccgca ccaacgcgca
     2461 gcccggactc ggtaatggcg cgcattgcgc ccagcgccat ctgatcgttg gcaaccagca
     2521 tcgcagtggg aacgatgccc tcattcagca tttgcatggt ttgttgaaaa ccggacatgg
     2581 cactccagtc gccttcccgt tccgctatcg gctgaatttg attgcgagtg agatatttat
     2641 gccagccagc cagacgcaga cgcgccgaga cagaacttaa tgggcccgct aacagcgcga
     2701 tttgctggtg acccaatgcg accagatgct ccacgcccag tcgcgtaccg tcttcatggg
     2761 agaaaataat actgttgatg ggtgtctggt cagagacatc aagaaataac gccggaacat
     2821 tagtgcaggc agcttccaca gcaatggcat cctggtcatc cagcggatag ttaatgatca
     2881 gcccactgac gcgttgcgcg agaagattgt gcaccgccgc tttacaggct tcgacgccgc
     2941 ttcgttctac catcgacacc accacgctgg cacccagttg atcggcgcga gatttaatcg
     3001 ccgcgacaat ttgcgacggc gcgtgcaggg ccagactgga ggtggcaacg ccaatcagca
     3061 acgactgttt gcccgccagt tgttgtgcca cgcggttggg aatgtaattc agctccgcca
     3121 tcgccgcttc cactttttcc cgcgttttcg cagaaacgtg gctggcctgg ttcaccacgc
     3181 gggaaacggt ctgataagag acaccggcat actctgcgac atcgtataac gttactggtt
     3241 tcacattcac caccctgaat tgactctctt ccgggcgcta tcatgccata ccgcgaaagg
     3301 ttttgcgcca ttcgatggtg tccgggatct cgacgctctc ccttatgcga
//
