LOCUS       sequence-431468-        2929 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fragmid fragment: (C' terminus) ZIM3 KRAB.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2929)
  AUTHORS   McGee AV, Liu YV, Griffith AL, Szegletes ZM, Wen B, Kraus C, Miller 
            NW, Steger RJ, Escude Velasco B, Bosch JA, Zirin JD, Viswanatha R, 
            Sontheimer EJ, Goodale A, Greene MA, Green TM, Doench JG
  TITLE     Modular vector assembly enables rapid assessment of emerging CRISPR 
            technologies.
  JOURNAL   Cell Genom. 2024 Mar 13;4(3):100519. doi: 
            10.1016/j.xgen.2024.100519.
  PUBMED    38484704
REFERENCE   2  (bases 1 to 2929)
  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"; journalName: "Cell 
            Genom."; date: "2024-03-13"; volume: "4(3):100519. doi"; pages: " 
            10.1016/j.xgen.2024.100519"
COMMENT     Sequence Label: AA092
FEATURES             Location/Qualifiers
     source          1..2929
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     364..386
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             468..515
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     primer_bind     complement(839..855)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(839..855)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(852..874)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    863..879
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(887..893,894..911,912..917))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    932..953
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1070..1087)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1241..1829)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1321..1340)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2007..2816)
                     /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="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     2730..2749
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2808..2827
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagagaatt cgagctcggt acctcgcgaa
      421 tacatctaga tgaagacggg tccgggagcg gcgggagcgg agggagcaaa aggccggcgg
      481 ccacgaaaaa ggccggccag gcaaaaaaga aaaagagcgg aggctccaac aacagccagg
      541 gcagagtgac cttcgaggac gtgaccgtga acttcaccca gggcgagtgg cagagactga
      601 accccgagca gagaaacctg tacagagatg tgatgctgga gaactacagc aacctggtga
      661 gcgtgggcca gggcgagacc accaagcccg acgtgatcct gagactggag cagggcaagg
      721 agccctggct ggaggaggag gaggtgctgg gcagcggcag agccgaggga agcacgtctt
      781 catcggatcc cgggcccgtc gactgcagag gcctgcatgc aagcttggtg taatcatggt
      841 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
      901 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
      961 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     1021 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     1081 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     1141 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     1201 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     1261 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     1321 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     1381 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     1441 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     1501 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     1561 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     1621 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     1681 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     1741 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     1801 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     1861 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     1921 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaagccca atctgaataa
     1981 tgttacaacc aattaaccaa ttctgattag aaaaactcat cgagcatcaa atgaaactgc
     2041 aatttattca tatcaggatt atcaatacca tatttttgaa aaagccgttt ctgtaatgaa
     2101 ggagaaaact caccgaggca gttccatagg atggcaagat cctggtatcg gtctgcgatt
     2161 ccgactcgtc caacatcaat acaacctatt aatttcccct cgtcaaaaat aaggttatca
     2221 agtgagaaat caccatgagt gacgactgaa tccggtgaga atggcaaaag tttatgcatt
     2281 tctttccaga cttgttcaac aggccagcca ttacgctcgt catcaaaatc actcgcatca
     2341 accaaaccgt tattcattcg tgattgcgcc tgagcgagac gaaatacgcg atcgctgtta
     2401 aaaggacaat tacaaacagg aatcgaatgc aaccggcgca ggaacactgc cagcgcatca
     2461 acaatatttt cacctgaatc aggatattct tctaatacct ggaatgctgt ttttccgggg
     2521 atcgcagtgg tgagtaacca tgcatcatca ggagtacgga taaaatgctt gatggtcgga
     2581 agaggcataa attccgtcag ccagtttagt ctgaccatct catctgtaac atcattggca
     2641 acgctacctt tgccatgttt cagaaacaac tctggcgcat cgggcttccc atacaagcga
     2701 tagattgtcg cacctgattg cccgacatta tcgcgagccc atttataccc atataaatca
     2761 gcatccatgt tggaatttaa tcgcggcctc gacgtttccc gttgaatatg gctcataaca
     2821 ccccttgtat tactgtttat gtaagcagac agttttattg ttcatgatga tatattttta
     2881 tcttgtgcaa tgtaacatca gagattttga gacacgggcc agagctgca
//
