LOCUS       sequence-368503-        2279 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Expresses a sgRNA to edit GLB1 gene
            .2_c.907A>G.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2279)
  TITLE     Gene editing is suitable to treat GM1 Gangliosidosis: a 
            proof-of-concept study
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 2279)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: BPK1520-sgRNA GLB1
FEATURES             Location/Qualifiers
     source          1..2279
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        208..456
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     208..228
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     379..398
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        477..552
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     rep_origin      complement(656..1244)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(736..755)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1415..2206,2207..2275))
                     /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     2038..2057
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(2276..2279,1..101))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
       61 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
      121 ccacctgacg tcgctagctg tacaaaaaag caggctttaa aggaaccaat tcagtcgact
      181 ggatccggta ccaaggtcgg gcaggaagag ggcctatttc ccatgattcc ttcatatttg
      241 catatacgat acaaggctgt tagagagata attagaatta atttgactgt aaacacaaag
      301 atattagtac aaaatacgtg acgtagaaag taataatttc ttgggtagtt tgcagtttta
      361 aaattatgtt ttaaaatgga ctatcatatg cttaccgtaa cttgaaagta tttcgatttc
      421 ttggctttat atatcttgtg gaaaggacga aacaccggcg agtatgaact tgtgaggttt
      481 tagagctaga aatagcaagt taaaataagg ctagtccgtt atcaacttga aaaagtggca
      541 ccgagtcggt gcttttttta agcttgggcc gctcgaggta cctctctaca tatgacatgt
      601 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
      661 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
      721 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
      781 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
      841 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
      901 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
      961 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     1021 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     1081 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     1141 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     1201 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     1261 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     1321 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     1381 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     1441 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     1501 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     1561 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     1621 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     1681 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     1741 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     1801 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     1861 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     1921 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     1981 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     2041 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     2101 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     2161 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     2221 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactc
//
