LOCUS       sequence-385670-        2490 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses U6-driven N7 sgRNA2 (poly-T mutated out), BsmbI dropout 
            cassette for cloning.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2490)
  AUTHORS   Tou CJ, Orr B, Kleinstiver BP
  TITLE     Precise cut-and-paste DNA insertion using engineered type V-K 
            CRISPR-associated transposases.
  JOURNAL   Nat Biotechnol. 2023 Jan 2. doi: 10.1038/s41587-022-01574-x.
  PUBMED    36593413
REFERENCE   2  (bases 1 to 2490)
  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: "Nat 
            Biotechnol. 2023 Jan 2. doi: 10.1038/s41587-022-01574-x."
COMMENT     Sequence Label: pU6_N7-sgRNA2
FEATURES             Location/Qualifiers
     source          1..2490
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        82..330
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     82..102
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     253..272
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     rep_origin      complement(741..1329)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(821..840)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1500..2291,2292..2360))
                     /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     2123..2142
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2361..2465)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacgtcgcta gctgtacaaa aaagcaggct ttaaaggaac caattcagtc gactggatcc
       61 ggtaccaagg tcgggcagga agagggccta tttcccatga ttccttcata tttgcatata
      121 cgatacaagg ctgttagaga gataattaga attaatttga ctgtaaacac aaagatatta
      181 gtacaaaata cgtgacgtag aaagtaataa tttcttgggt agtttgcagt tttaaaatta
      241 tgttttaaaa tggactatca tatgcttacc gtaacttgaa agtatttcga tttcttggct
      301 ttatatatct tgtggaaagg acgaaacacc gatattctta taacagcgcc gcagttcatg
      361 ctttcttaag ccaatgtact gtgaaaaatc tgggttagtt tggcggttgg aaggccgtca
      421 tgctttctga cccttgtagc tgcccgcttc tgatgctgcc atctttagaa ttctataggt
      481 gggataggtg cgctcccagc aataaggagt aaggcttata gctatagccg ttattcataa
      541 cggtgcggat taccacagtg gtggctactg aatcaccccc ttcgtcgggg gaaccctcca
      601 aaaggtgggt tgaaagggag acgattaatg cgtctccttt ttttaagctt gggccgctcg
      661 aggtacctct ctacatatga catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
      721 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
      781 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
      841 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
      901 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
      961 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     1021 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     1081 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     1141 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     1201 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     1261 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     1321 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     1381 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     1441 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     1501 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     1561 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     1621 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     1681 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     1741 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     1801 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     1861 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     1921 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     1981 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     2041 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     2101 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     2161 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     2221 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     2281 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     2341 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     2401 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     2461 ccgcgcacat ttccccgaaa agtgccacct
//
