LOCUS       sequence-421622-        2982 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  U6-JAGBLH010000220 Cas8-HNH DR-BpiI-DR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2982)
  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 2982)
  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: pHS1871
FEATURES             Location/Qualifiers
     source          1..2982
                     /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     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(1132..1149)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1132..1148)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1141..1163)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(1357..1376)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1476..1498
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1536..1554)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1622..1726
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1727..1795,1796..2587)
                     /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(1945..1964)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(2758..2982,1..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 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
      541 ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag
      601 ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag
      661 ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg
      721 tggaattgtg agcggataac aatttcacgc 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 gaaacaccgt gtaccgccgg ataggcggtt tagaagatgt cttcctggga cgaagacaag
     1081 tgtaccgccg gataggcggt ttagaagttt tttggtaccg agctcgaatt cactggccgt
     1141 cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc
     1201 acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca
     1261 acagttgcgc agcctgaatg gcgaatggcg cctgatgcgg tattttctcc ttacgcatct
     1321 gtgcggtatt tcacaccgca tatggtgcac tctcagtaca atctgctctg atgccgcata
     1381 gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct
     1441 cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt
     1501 ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc tatttttata
     1561 ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt
     1621 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     1681 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     1741 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     1801 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     1861 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     1921 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     1981 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     2041 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     2101 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     2161 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     2221 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     2281 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     2341 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     2401 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     2461 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     2521 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     2581 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     2641 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     2701 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     2761 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     2821 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     2881 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     2941 gaactctgta gcaccgccta catacctcgc tctgctaatc ct
//
