LOCUS       sequence-351753-        3837 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of human codon-optimized NlaCas11 with N terminal NLS and
            HA tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3837)
  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 3837)
  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-EF1a-Nla-cas11
FEATURES             Location/Qualifiers
     source          1..3837
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(32..51)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        201..1363
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          432..1354
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1311..1331
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      1369..1378
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1381..1401
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1402..1428
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(1837..1854)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1843..1954
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     terminator      2112..2141
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      2163..2750
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2651..2670
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2762..2789)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(2810..3625)
                     /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     3533..3552
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(3626..3728)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(3729..3760)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 gatctcccga tcccctatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
       61 gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt
      121 aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt agggttaggc
      181 gttttgcgct gcttcgcgga ggctccggtg cccgtcagtg ggcagagcgc acatcgccaa
      241 cagtccccga gaagttgggg ggaggggtcg gcaattgaac cggtgcctag agaaggtgga
      301 gcggggtaaa ctgggaaagt gatgtcgtgt actggctccg cctttttccc gagggtgggg
      361 gagaaccgta tataagtgca gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg
      421 ccagaacaca ggtaagtgcc gtgtgtggtt cccgcgggcc tggcctcttt acgggttatg
      481 gcccttgcgt gccttgaatt acttccacct ggctccagta cgtgattctt gatcccgagc
      541 tggagccagg ggcgggcctt gcgctttagg agccccttcg cctcgtgctt gagttgaggc
      601 ctggcctggg cgctggggcc gccgcgtgcg aatctggtgg caccttcgcg cctgtctcgc
      661 tgctttcgat aagtctctag ccatttaaaa tttttgatga cctgctgcga cgcttttttt
      721 ctggcaagat agtcttgtaa atgcgggcca ggatctgcac actggtattt cggtttttgg
      781 ggccgcgggc ggcgacgggg cccgtgcgtc ccagcgcaca tgttcggaga ggcggggcct
      841 gcgagcgcgg ccaccgagaa tcggacgggg gtagtctcaa gctggccggc ctgctctggt
      901 gcctggcctc gcgcagccgt gtatcgcccc gcactgggcg gcaaggctgg cccggtcggc
      961 accagttgcg tgagcggaaa gatggccgct tcccggccct gctccagggg gctcaaaatg
     1021 gaggacgcgg cgctcgggag agcgggcggg tgagtcaccc acacaaagga aaggggcctt
     1081 tccgtcctca gccgtcgctt catgtgactc cacggagtac cgggcgccgt ccaggcacct
     1141 cgattagttc tggagctttt ggagtacgtc gtctttaggt tggggggagg ggttttatgc
     1201 gatggagttt ccccacactg agtgggtgga gactgaagtt aggccagctt ggcacttgat
     1261 gtaattctcc ttggaatttg ccctttttga gtttggatct tggttcattc tcaagcctca
     1321 gacagtggtt caaagttttt ttcttccatt tcaggtgtcg tgaacaccgc caccatggtg
     1381 cccaagaaaa agcggaaggt gtacccctac gacgtgcccg attatgccgg ctctgtggga
     1441 ggcctcgacc ggaacagaca ggatatcggc tatgtgctgg gcagactgtt cgccgtgctg
     1501 gaaaagattc aggccgaggc caatcctggc ctgaacgcca caatcgccga cagatatttt
     1561 ggcagcgcca gcagcacacc tatcgccgtg tttggcaccc tgatgagact gctgcctcac
     1621 cacctgaaca agctggaatt cgagggcaga gccgtgcagc tccagtggga gatcagacag
     1681 atcctggaac actgccagcg gttccccaat cacctgaacc tggaacagca gggactgttt
     1741 gccatcggct actaccacga gacacagttt ctgttcacca aggacgccct gaagaacctg
     1801 ttcaacgagg ccaagaccgc ctgactcgct gatcagcctc gactgtgcct tctagttgcc
     1861 agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca
     1921 ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta
     1981 ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac aatagcagga
     2041 acgaattcaa gcttgatatc attcaggacg agcctcagac tccagcgtaa ctggactgca
     2101 atcaactcac tggctcacct tcacgggtgg gcctttcttc ggtagaaaat caaaggatct
     2161 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     2221 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgag gtaactggct
     2281 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     2341 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     2401 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     2461 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     2521 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     2581 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     2641 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     2701 ttgagcatcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     2761 acgcagaaag gcccacccga aggtgagcca ggtgattaca tttgggccct cagaaaaact
     2821 catcgagcat caagtgaaac tgcaatttat tcatatcagg attatcaata ccatattttt
     2881 gaaaaagccg tttctgtaat gaaggagaaa actcaccgag gcagttccat aggatggcaa
     2941 gatcctggta tcggtctgcg attccgactc gtccaacatc aatacaacct attaatttcc
     3001 cctcgtcaaa aataaggtta tcaagtgaga aatcaccatg agtgacgact gaatccggtg
     3061 agaatggcaa aagcttatgc atttctttcc agacttgttc aacaggccag ccattacgct
     3121 cgtcatcaaa atcactcgca ccaaccaaac cgttattcat tcgtgattgc gcctgagcga
     3181 gacgaaatac gcgatcgccg ttaaaaggac aattacaaac aggaatcgaa tgcaaccggc
     3241 gcaggaacac tgccagcgca tcaacaatat tttcacctga atcaggatat tcttctaata
     3301 cctggaatgc tgttttccct gggatcgcag tggtgagtaa ccatgcatca tcaggagtac
     3361 ggataaaatg cttgatggtc ggaagaggca taaattccgt cagccagttt agcctgacca
     3421 tctcatctgt aacatcattg gcaacgctac ctttgccatg tttcagaaac aactctggcg
     3481 catcgggctt cccatacaat cgatagattg tcgcacctga ttgcccgaca ttatcgcgag
     3541 cccatttata cccatataaa tcagcatcca tgttggaatt taatcgcggc ctcgagcaag
     3601 acgtttcccg ttgaatatgg ctcattttag cttccttagc tcctgaaaat ctcgataact
     3661 caaaaaatac gcccggtagt gatcttattt cattatggtg aaagttggaa cctcttacgt
     3721 gccgatcaag tcaaaagcct ccggtcggag gcttttgact ttctgctatg gaggtcaggt
     3781 atgatttaaa tggtcagtat tgagcgatat ctagagaatt cgtccgacgg atcggga
//
