LOCUS       sequence-410160-        3645 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses AcrIC9 with NLS in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3645)
  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 3645)
  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-AcrIC9-NLS
FEATURES             Location/Qualifiers
     source          1..3645
                     /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             1622..1642
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(1658..1675)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1664..1775
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     terminator      1933..1962
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      1984..2571
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2472..2491
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2583..2610)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(2631..3446)
                     /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     3354..3373
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(3447..3549)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(3550..3581)
                     /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 accagcttct acaagatcac cgcatacaac tctcaggccc tgtacttctg
     1441 gggcaccgac gccgatgtgg acagatatgt ggactggctg aaccgggacc gcgagatcaa
     1501 tgtgtatgcc gccgaggcca ttcctgaggc cgaatgggct caatacgagg gcagagatga
     1561 tgtgctgagc ggcgaggaat gcggctggga cgatttcatg tctgccgaag ctggcggcag
     1621 ccccaagaaa aagcggaaag tgtgactcgc tgatcagcct cgactgtgcc ttctagttgc
     1681 cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc
     1741 actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct
     1801 attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg
     1861 aacgaattca agcttgatat cattcaggac gagcctcaga ctccagcgta actggactgc
     1921 aatcaactca ctggctcacc ttcacgggtg ggcctttctt cggtagaaaa tcaaaggatc
     1981 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     2041 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga ggtaactggc
     2101 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     2161 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     2221 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     2281 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     2341 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     2401 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     2461 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     2521 cttgagcatc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     2581 aacgcagaaa ggcccacccg aaggtgagcc aggtgattac atttgggccc tcagaaaaac
     2641 tcatcgagca tcaagtgaaa ctgcaattta ttcatatcag gattatcaat accatatttt
     2701 tgaaaaagcc gtttctgtaa tgaaggagaa aactcaccga ggcagttcca taggatggca
     2761 agatcctggt atcggtctgc gattccgact cgtccaacat caatacaacc tattaatttc
     2821 ccctcgtcaa aaataaggtt atcaagtgag aaatcaccat gagtgacgac tgaatccggt
     2881 gagaatggca aaagcttatg catttctttc cagacttgtt caacaggcca gccattacgc
     2941 tcgtcatcaa aatcactcgc accaaccaaa ccgttattca ttcgtgattg cgcctgagcg
     3001 agacgaaata cgcgatcgcc gttaaaagga caattacaaa caggaatcga atgcaaccgg
     3061 cgcaggaaca ctgccagcgc atcaacaata ttttcacctg aatcaggata ttcttctaat
     3121 acctggaatg ctgttttccc tgggatcgca gtggtgagta accatgcatc atcaggagta
     3181 cggataaaat gcttgatggt cggaagaggc ataaattccg tcagccagtt tagcctgacc
     3241 atctcatctg taacatcatt ggcaacgcta cctttgccat gtttcagaaa caactctggc
     3301 gcatcgggct tcccatacaa tcgatagatt gtcgcacctg attgcccgac attatcgcga
     3361 gcccatttat acccatataa atcagcatcc atgttggaat ttaatcgcgg cctcgagcaa
     3421 gacgtttccc gttgaatatg gctcatttta gcttccttag ctcctgaaaa tctcgataac
     3481 tcaaaaaata cgcccggtag tgatcttatt tcattatggt gaaagttgga acctcttacg
     3541 tgccgatcaa gtcaaaagcc tccggtcgga ggcttttgac tttctgctat ggaggtcagg
     3601 tatgatttaa atggtcagta ttgagcgata tctagagaat tcgtc
//
