LOCUS       sequence-351755-        2163 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Vector for cloning Nla-IC-CRISPR RNA for expression in mammalian 
            cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2163)
  AUTHORS   Tan R, Krueger RK, Gramelspacher MJ, Zhou X, Xiao Y, Ke A, Hou Z, 
            Zhang Y
  TITLE     Cas11 enables genome engineering in human cells with compact 
            CRISPR-Cas3 systems.
  JOURNAL   Mol Cell. 2022 Feb 17;82(4):852-867.e5. doi: 
            10.1016/j.molcel.2021.12.032. Epub 2022 Jan 19.
  PUBMED    35051351
REFERENCE   2  (bases 1 to 2163)
  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"; doi: 
            "10.1016/j.molcel.2021.12.032"; journalName: "Mol Cell"; date: 
            "2022-02-17- 17"; volume: "82"; issue: "4"; pages: "852-867.e5"
COMMENT     Sequence Label: pSmart HC Kan-U6-Nla-repeat-BbsI-Nla-repeat
FEATURES             Location/Qualifiers
     source          1..2163
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        42..290
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     42..62
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     213..232
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     terminator      443..474
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        475..577
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             578..1393
                     /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
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNGDRVFRLAQAQSRMNNGLVGA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(651..670)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      1414..1441
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(1453..2040)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1533..1552)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2062..2091)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
ORIGIN
        1 ccaagctcgg atccgatatc gacgtcaagg tcgggcagga agagggccta tttcccatga
       61 ttccttcata tttgcatata cgatacaagg ctgttagaga gataattaga attaatttga
      121 ctgtaaacac aaagatatta gtacaaaata cgtgacgtag aaagtaataa tttcttgggt
      181 agtttgcagt tttaaaatta tgttttaaaa tggactatca tatgcttacc gtaacttgaa
      241 agtatttcga tttcttggct ttatatatct tgtggaaagg acgaaacacc gtcagccgcc
      301 tctaggcggc tgtgtgttga aacgggtctt cattattcca gaagacaatc agccgcctct
      361 aggcggctgt gtgttgaaac ttttttttct agatatcgct caatactgac catttaaatc
      421 atacctgacc tccatagcag aaagtcaaaa gcctccgacc ggaggctttt gacttgatcg
      481 gcacgtaaga ggttccaact ttcaccataa tgaaataaga tcactaccgg gcgtattttt
      541 tgagttatcg agattttcag gagctaagga agctaaaatg agccatattc aacgggaaac
      601 gtcttgctcg aggccgcgat taaattccaa catggatgct gatttatatg ggtataaatg
      661 ggctcgcgat aatgtcgggc aatcaggtgc gacaatctat cgattgtatg ggaagcccga
      721 tgcgccagag ttgtttctga aacatggcaa aggtagcgtt gccaatgatg ttacagatga
      781 gatggtcagg ctaaactggc tgacggaatt tatgcctctt ccgaccatca agcattttat
      841 ccgtactcct gatgatgcat ggttactcac cactgcgatc ccagggaaaa cagcattcca
      901 ggtattagaa gaatatcctg attcaggtga aaatattgtt gatgcgctgg cagtgttcct
      961 gcgccggttg cattcgattc ctgtttgtaa ttgtcctttt aacggcgatc gcgtatttcg
     1021 tctcgctcag gcgcaatcac gaatgaataa cggtttggtt ggtgcgagtg attttgatga
     1081 cgagcgtaat ggctggcctg ttgaacaagt ctggaaagaa atgcataagc ttttgccatt
     1141 ctcaccggat tcagtcgtca ctcatggtga tttctcactt gataacctta tttttgacga
     1201 ggggaaatta ataggttgta ttgatgttgg acgagtcgga atcgcagacc gataccagga
     1261 tcttgccatc ctatggaact gcctcggtga gttttctcct tcattacaga aacggctttt
     1321 tcaaaaatat ggtattgata atcctgatat gaataaattg cagtttcact tgatgctcga
     1381 tgagtttttc tgagggccca aatgtaatca cctggctcac cttcgggtgg gcctttctgc
     1441 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgatgctc
     1501 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     1561 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     1621 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     1681 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     1741 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     1801 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     1861 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     1921 gaagccagtt acctcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     1981 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     2041 gaagatcctt tgattttcta ccgaagaaag gcccacccgt gaaggtgagc cagtgagttg
     2101 attgcagtcc agttacgctg gagtctgagg ctcgtcctga atgatatcaa gcttgaattc
     2161 gtt
//
