LOCUS       sequence-410143-        3657 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses AcrIC8 with NLS in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3657)
  AUTHORS   Hu C, Myers MT, Zhou X, Hou Z, Lozen ML, Nam KH, Zhang Y, Ke A
  TITLE     Exploiting activation and inactivation mechanisms in type I-C 
            CRISPR-Cas3 for genome-editing applications.
  JOURNAL   Mol Cell. 2024 Feb 1;84(3):463-475.e5. doi: 
            10.1016/j.molcel.2023.12.034. Epub 2024 Jan 18.
  PUBMED    38242128
REFERENCE   2  (bases 1 to 3657)
  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.2023.12.034"; journalName: "Mol Cell"; date: 
            "2024-02-1- 1"; volume: "84"; issue: "3"; pages: "463-475.e5"
COMMENT     Sequence Label: pSmart-EF1alpha-AcrIC8-NLS
FEATURES             Location/Qualifiers
     source          1..3657
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(45..64)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        214..1376
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          445..1367
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1324..1344
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             1634..1654
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(1670..1687)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1676..1787
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     terminator      1945..1974
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      1996..2583
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2484..2503
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2595..2622)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(2643..3458)
                     /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     3366..3385
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(3459..3561)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(3562..3593)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 cgacggatcg ggagatctcc cgatccccta tggtgcactc tcagtacaat ctgctctgat
       61 gccgcatagt taagccagta tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc
      121 gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg acaattgcat gaagaatctg
      181 cttagggtta ggcgttttgc gctgcttcgc ggaggctccg gtgcccgtca gtgggcagag
      241 cgcacatcgc caacagtccc cgagaagttg gggggagggg tcggcaattg aaccggtgcc
      301 tagagaaggt ggagcggggt aaactgggaa agtgatgtcg tgtactggct ccgccttttt
      361 cccgagggtg ggggagaacc gtatataagt gcagtagtcg ccgtgaacgt tctttttcgc
      421 aacgggtttg ccgccagaac acaggtaagt gccgtgtgtg gttcccgcgg gcctggcctc
      481 tttacgggtt atggcccttg cgtgccttga attacttcca cctggctcca gtacgtgatt
      541 cttgatcccg agctggagcc aggggcgggc cttgcgcttt aggagcccct tcgcctcgtg
      601 cttgagttga ggcctggcct gggcgctggg gccgccgcgt gcgaatctgg tggcaccttc
      661 gcgcctgtct cgctgctttc gataagtctc tagccattta aaatttttga tgacctgctg
      721 cgacgctttt tttctggcaa gatagtcttg taaatgcggg ccaggatctg cacactggta
      781 tttcggtttt tggggccgcg ggcggcgacg gggcccgtgc gtcccagcgc acatgttcgg
      841 agaggcgggg cctgcgagcg cggccaccga gaatcggacg ggggtagtct caagctggcc
      901 ggcctgctct ggtgcctggc ctcgcgcagc cgtgtatcgc cccgcactgg gcggcaaggc
      961 tggcccggtc ggcaccagtt gcgtgagcgg aaagatggcc gcttcccggc cctgctccag
     1021 ggggctcaaa atggaggacg cggcgctcgg gagagcgggc gggtgagtca cccacacaaa
     1081 ggaaaggggc ctttccgtcc tcagccgtcg cttcatgtga ctccacggag taccgggcgc
     1141 cgtccaggca cctcgattag ttctggagct tttggagtac gtcgtcttta ggttgggggg
     1201 aggggtttta tgcgatggag tttccccaca ctgagtgggt ggagactgaa gttaggccag
     1261 cttggcactt gatgtaattc tccttggaat ttgccctttt tgagtttgga tcttggttca
     1321 ttctcaagcc tcagacagtg gttcaaagtt tttttcttcc atttcaggtg tcgtgaacac
     1381 cgccaccatg tacgccatcc ggaagatcca gttcttctac ggccccaccg acaagaagtc
     1441 ctacgttgga gaagaggctg gcggcagaag agagctgttc aagacaagag ccgaggctca
     1501 ggccagaatc gaggatctgg aagagggcgt gtactatctg gcccacaacg agagcggcag
     1561 acccgactat aagatcgtgt gggttcgagg cgaggcccag tttgaacacg ccagatggat
     1621 gagaggcggc agccccaaga aaaagcggaa agtgtgactc gctgatcagc ctcgactgtg
     1681 ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa
     1741 ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt
     1801 aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa
     1861 gacaatagca ggaacgaatt caagcttgat atcattcagg acgagcctca gactccagcg
     1921 taactggact gcaatcaact cactggctca ccttcacggg tgggcctttc ttcggtagaa
     1981 aatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     2041 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     2101 gaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     2161 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     2221 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     2281 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     2341 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     2401 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     2461 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     2521 cgccacctct gacttgagca tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     2581 aaaaacgcca gcaacgcaga aaggcccacc cgaaggtgag ccaggtgatt acatttgggc
     2641 cctcagaaaa actcatcgag catcaagtga aactgcaatt tattcatatc aggattatca
     2701 ataccatatt tttgaaaaag ccgtttctgt aatgaaggag aaaactcacc gaggcagttc
     2761 cataggatgg caagatcctg gtatcggtct gcgattccga ctcgtccaac atcaatacaa
     2821 cctattaatt tcccctcgtc aaaaataagg ttatcaagtg agaaatcacc atgagtgacg
     2881 actgaatccg gtgagaatgg caaaagctta tgcatttctt tccagacttg ttcaacaggc
     2941 cagccattac gctcgtcatc aaaatcactc gcaccaacca aaccgttatt cattcgtgat
     3001 tgcgcctgag cgagacgaaa tacgcgatcg ccgttaaaag gacaattaca aacaggaatc
     3061 gaatgcaacc ggcgcaggaa cactgccagc gcatcaacaa tattttcacc tgaatcagga
     3121 tattcttcta atacctggaa tgctgttttc cctgggatcg cagtggtgag taaccatgca
     3181 tcatcaggag tacggataaa atgcttgatg gtcggaagag gcataaattc cgtcagccag
     3241 tttagcctga ccatctcatc tgtaacatca ttggcaacgc tacctttgcc atgtttcaga
     3301 aacaactctg gcgcatcggg cttcccatac aatcgataga ttgtcgcacc tgattgcccg
     3361 acattatcgc gagcccattt atacccatat aaatcagcat ccatgttgga atttaatcgc
     3421 ggcctcgagc aagacgtttc ccgttgaata tggctcattt tagcttcctt agctcctgaa
     3481 aatctcgata actcaaaaaa tacgcccggt agtgatctta tttcattatg gtgaaagttg
     3541 gaacctctta cgtgccgatc aagtcaaaag cctccggtcg gaggcttttg actttctgct
     3601 atggaggtca ggtatgattt aaatggtcag tattgagcga tatctagaga attcgtc
//
