LOCUS       sequence-415895-        3675 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses AcrIE2 with NLS in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3675)
  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 3675)
  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-AcrIE2-NLS
FEATURES             Location/Qualifiers
     source          1..3675
                     /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"
     regulatory      1382..1391
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1652..1672
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(1688..1705)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1694..1805
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     terminator      1963..1992
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      2014..2601
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2502..2521
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2613..2640)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(2661..3476)
                     /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     3384..3403
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(3477..3579)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(3580..3611)
                     /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 gccaacacct acctgatcga ccccagaaag aacaacgaca acagcggcga
     1441 gcggttcacc gtggatgccg tggatattac agccgccgct aagtctgccg ctcagcagat
     1501 tctgggcgaa gagttcgaag gcctggtgta cagagagaca ggcgagtcta acggcagcgg
     1561 catgtttcag gcctaccacc atctgcacgg caccaacaga accgagacaa ccgtgggcta
     1621 ccccttccac gtgatggaac tgggcggcag ccccaagaaa aagcggaaag tgtgactcgc
     1681 tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg
     1741 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt
     1801 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc
     1861 aagggggagg attgggaaga caatagcagg aacgaattca agcttgatat cattcaggac
     1921 gagcctcaga ctccagcgta actggactgc aatcaactca ctggctcacc ttcacgggtg
     1981 ggcctttctt cggtagaaaa tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     2041 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     2101 gctaccaact ctttttccga ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     2161 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     2221 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     2281 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     2341 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     2401 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     2461 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     2521 tatagtcctg tcgggtttcg ccacctctga cttgagcatc gatttttgtg atgctcgtca
     2581 ggggggcgga gcctatggaa aaacgccagc aacgcagaaa ggcccacccg aaggtgagcc
     2641 aggtgattac atttgggccc tcagaaaaac tcatcgagca tcaagtgaaa ctgcaattta
     2701 ttcatatcag gattatcaat accatatttt tgaaaaagcc gtttctgtaa tgaaggagaa
     2761 aactcaccga ggcagttcca taggatggca agatcctggt atcggtctgc gattccgact
     2821 cgtccaacat caatacaacc tattaatttc ccctcgtcaa aaataaggtt atcaagtgag
     2881 aaatcaccat gagtgacgac tgaatccggt gagaatggca aaagcttatg catttctttc
     2941 cagacttgtt caacaggcca gccattacgc tcgtcatcaa aatcactcgc accaaccaaa
     3001 ccgttattca ttcgtgattg cgcctgagcg agacgaaata cgcgatcgcc gttaaaagga
     3061 caattacaaa caggaatcga atgcaaccgg cgcaggaaca ctgccagcgc atcaacaata
     3121 ttttcacctg aatcaggata ttcttctaat acctggaatg ctgttttccc tgggatcgca
     3181 gtggtgagta accatgcatc atcaggagta cggataaaat gcttgatggt cggaagaggc
     3241 ataaattccg tcagccagtt tagcctgacc atctcatctg taacatcatt ggcaacgcta
     3301 cctttgccat gtttcagaaa caactctggc gcatcgggct tcccatacaa tcgatagatt
     3361 gtcgcacctg attgcccgac attatcgcga gcccatttat acccatataa atcagcatcc
     3421 atgttggaat ttaatcgcgg cctcgagcaa gacgtttccc gttgaatatg gctcatttta
     3481 gcttccttag ctcctgaaaa tctcgataac tcaaaaaata cgcccggtag tgatcttatt
     3541 tcattatggt gaaagttgga acctcttacg tgccgatcaa gtcaaaagcc tccggtcgga
     3601 ggcttttgac tttctgctat ggaggtcagg tatgatttaa atggtcagta ttgagcgata
     3661 tctagagaat tcgtc
//
