LOCUS       sequence-416257-        2985 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  U6-MWEE01000013 Cas5-HNH DR-BpiI-DR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2985)
  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 2985)
  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: pHS1528
FEATURES             Location/Qualifiers
     source          1..2985
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     265..284
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     518..535
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    652..673
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(688..693,694..711,712..718)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    726..742
                     /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)."
     primer_bind     731..753
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     750..766
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     750..766
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        780..1028
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     780..800
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     951..970
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(1135..1152)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1135..1151)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1144..1166)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(1360..1379)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1479..1501
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1539..1557)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1625..1729
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1730..1798,1799..2590)
                     /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     complement(1948..1967)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(2761..2985,1..364)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
       61 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
      121 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
      181 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
      241 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
      301 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
      361 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
      421 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
      481 agctgatacc tctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
      541 ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag
      601 ctggcacgac aggtttcccg actggaaagc gggcagtgaa cgcaacgcaa ttaatgtgag
      661 ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg
      721 tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg attacgccag
      781 agggcctatt tcccatgatt ccttcatatt tgcatatacg atacaaggct gttagagaga
      841 taattggaat taatttgact gtaaacacaa agatattagt acaaaatacg tgacgtagaa
      901 agtaataatt tcttgggtag tttgcagttt taaaattatg ttttaaaatg gactatcata
      961 tgcttaccgt aacttgaaag tatttcgatt tcttggcttt atatatcttg tggaaaggac
     1021 gaaacaccgg tattccccac gcatgtgggg gtgaaccgat gtcttcctgg gacgaagaca
     1081 agtattcccc acgcatgtgg gggtgaaccg ttttttggta ccgagctcga attcactggc
     1141 cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc
     1201 agcacatccc cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc
     1261 ccaacagttg cgcagcctga atggcgaatg gcgcctgatg cggtattttc tccttacgca
     1321 tctgtgcggt atttcacacc gcatatggtg cactctcagt acaatctgct ctgatgccgc
     1381 atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
     1441 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
     1501 gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt
     1561 ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa
     1621 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     1681 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     1741 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     1801 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     1861 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     1921 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     1981 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     2041 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     2101 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     2161 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     2221 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     2281 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     2341 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     2401 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     2461 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     2521 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     2581 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     2641 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     2701 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     2761 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     2821 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     2881 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     2941 caagaactct gtagcaccgc ctacatacct cgctctgcta atcct
//
