LOCUS       sequence-416248-        3363 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  NZ_CP025815 DinG HNH crRNA expression in pCOLADuet-1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3363)
  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 3363)
  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: pHS1182
FEATURES             Location/Qualifiers
     source          1..3363
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     97..116
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        97..115
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      219..266
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(496..1311)
                     /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(496..1311)
                     /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     1219..1238
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(1312..1403)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1421..2056)
                     /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(1566..1585)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2254..3336)
                     /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     3298..3317
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(join(3337..3363,1..51))
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 cgggcgctat catgccatac cgcgaaaggt tttgcgccat tcgatggtgt ccgggatctc
       61 gacgctctcc cttatgcgac tcctgcatta ggaaattaat acgactcact atagggaata
      121 cagccatccg gaaattgggt ggctactaaa gggagaagtc atttaataag gccactgtta
      181 aaaatacagc catccggaaa ttgggtggct actaaaggct agcataaccc cttggggcct
      241 ctaaacgggt cttgaggggt tttttgctga aacctcaggc atttgagaag cacacggtca
      301 cactgcttcc ggtagtcaat aaaccggtaa accagcaata gacataagcg gctatttaac
      361 gaccctgccc tgaaccgacg acaagctgac gaccgggtct ccgcaagtgg cacttttcgg
      421 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
      481 ctcatgaatt aattcttaga aaaactcatc gagcatcaaa tgaaactgca atttattcat
      541 atcaggatta tcaataccat atttttgaaa aagccgtttc tgtaatgaag gagaaaactc
      601 accgaggcag ttccatagga tggcaagatc ctggtatcgg tctgcgattc cgactcgtcc
      661 aacatcaata caacctatta atttcccctc gtcaaaaata aggttatcaa gtgagaaatc
      721 accatgagtg acgactgaat ccggtgagaa tggcaaaagt ttatgcattt ctttccagac
      781 ttgttcaaca ggccagccat tacgctcgtc atcaaaatca ctcgcatcaa ccaaaccgtt
      841 attcattcgt gattgcgcct gagcgagacg aaatacgcgg tcgctgttaa aaggacaatt
      901 acaaacagga atcgaatgca accggcgcag gaacactgcc agcgcatcaa caatattttc
      961 acctgaatca ggatattctt ctaatacctg gaatgctgtt ttcccgggga tcgcagtggt
     1021 gagtaaccat gcatcatcag gagtacggat aaaatgcttg atggtcggaa gaggcataaa
     1081 ttccgtcagc cagtttagtc tgaccatctc atctgtaaca tcattggcaa cgctaccttt
     1141 gccatgtttc agaaacaact ctggcgcatc gggcttccca tacaatcgat agattgtcgc
     1201 acctgattgc ccgacattat cgcgagccca tttataccca tataaatcag catccatgtt
     1261 ggaatttaat cgcggcctag agcaagacgt ttcccgttga atatggctca tactcttcct
     1321 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     1381 atgtatttag aaaaataaac aaataggcat gctagcgcag aaacgtccta gaagatgcca
     1441 ggaggatact tagcagagag acaataaggc cggagcgaag ccgtttttcc ataggctccg
     1501 cccccctgac gaacatcacg aaatctgacg ctcaaatcag tggtggcgaa acccgacagg
     1561 actataaaga taccaggcgt ttccccctga tggctccctc ttgcgctctc ctgttcccgt
     1621 cctgcggcgt ccgtgttgtg gtggaggctt tacccaaatc accacgtccc gttccgtgta
     1681 gacagttcgc tccaagctgg gctgtgtgca agaacccccc gttcagcccg actgctgcgc
     1741 cttatccggt aactatcatc ttgagtccaa cccggaaaga cacgacaaaa cgccactggc
     1801 agcagccatt ggtaactgag aattagtgga tttagatatc gagagtcttg aagtggtggc
     1861 ctaacagagg ctacactgaa aggacagtat ttggtatctg cgctccacta aagccagtta
     1921 ccaggttaag cagttcccca actgacttaa ccttcgatca aaccgcctcc ccaggcggtt
     1981 ttttcgttta cagagcagga gattacgacg atcgtaaaag gatctcaaga agatccttta
     2041 cggattcccg acaccatcac tctagatttc agtgcaattt atctcttcaa atgtagcacc
     2101 tgaagtcagc cccatacgat ataagttgta attctcatgt tagtcatgcc ccgcgcccac
     2161 cggaaggagc tgactgggtt gaaggctctc aagggcatcg gtcgagatcc cggtgcctaa
     2221 tgagtgagct aacttacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     2281 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     2341 gggcgccagg gtggtttttc ttttcaccag tgagacgggc aacagctgat tgcccttcac
     2401 cgcctggccc tgagagagtt gcagcaagcg gtccacgctg gtttgcccca gcaggcgaaa
     2461 atcctgtttg atggtggtta acggcgggat ataacatgag ctgtcttcgg tatcgtcgta
     2521 tcccactacc gagatgtccg caccaacgcg cagcccggac tcggtaatgg cgcgcattgc
     2581 gcccagcgcc atctgatcgt tggcaaccag catcgcagtg ggaacgatgc cctcattcag
     2641 catttgcatg gtttgttgaa aaccggacat ggcactccag tcgccttccc gttccgctat
     2701 cggctgaatt tgattgcgag tgagatattt atgccagcca gccagacgca gacgcgccga
     2761 gacagaactt aatgggcccg ctaacagcgc gatttgctgg tgacccaatg cgaccagatg
     2821 ctccacgccc agtcgcgtac cgtcttcatg ggagaaaata atactgttga tgggtgtctg
     2881 gtcagagaca tcaagaaata acgccggaac attagtgcag gcagcttcca cagcaatggc
     2941 atcctggtca tccagcggat agttaatgat cagcccactg acgcgttgcg cgagaagatt
     3001 gtgcaccgcc gctttacagg cttcgacgcc gcttcgttct accatcgaca ccaccacgct
     3061 ggcacccagt tgatcggcgc gagatttaat cgccgcgaca atttgcgacg gcgcgtgcag
     3121 ggccagactg gaggtggcaa cgccaatcag caacgactgt ttgcccgcca gttgttgtgc
     3181 cacgcggttg ggaatgtaat tcagctccgc catcgccgct tccacttttt cccgcgtttt
     3241 cgcagaaacg tggctggcct ggttcaccac gcgggaaacg gtctgataag agacaccggc
     3301 atactctgcg acatcgtata acgttactgg tttcacattc accaccctga attgactctc
     3361 ttc
//
