LOCUS       sequence-214544-        2116 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Derivative of pTargetF with gRNA targeting FRT site.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2116)
  AUTHORS   Swings T, Marciano DC, Atri B, Bosserman RE, Wang C, Leysen M, Bonte
            C, Schalck T, Furey I, Van den Bergh B, Verstraeten N, Christie PJ, 
            Herman C, Lichtarge O, Michiels J
  TITLE     CRISPR-FRT targets shared sites in a knock-out collection for 
            off-the-shelf genome editing.
  JOURNAL   Nat Commun. 2018 Jun 8;9(1):2231. doi: 10.1038/s41467-018-04651-5.
  PUBMED    29884781
REFERENCE   2  (bases 1 to 2116)
  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: "Nat 
            Commun."; date: "2018-06-8"; volume: "9(1):2231. doi"; pages: " 
            10.1038/s41467-018-04651-5"
COMMENT     Sequence Label: pTargetF-FRT
FEATURES             Location/Qualifiers
     source          1..2116
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             124..915
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     rep_origin      1090..1678
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1579..1598
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1832..1849
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        1926..1960
                     /label=J23119(SpeI) promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119) modified to end with an SpeI site"
     misc_RNA        1979..2054
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
ORIGIN
        1 tcgagttcat gtgcagctcc atcagcaaaa ggggatgata agtttatcac caccgactat
       61 ttgcaacagt gccgttgatc gtgctatgat cgactgatgt catcagcggt ggagtgcaat
      121 gtcatgaggg aagcggtgat cgccgaagta tcgactcaac tatcagaggt agttggcgtc
      181 atcgagcgcc atctcgaacc gacgttgctg gccgtacatt tgtacggctc cgcagtggat
      241 ggcggcctga agccacacag tgatattgat ttgctggtta cggtgaccgt aaggcttgat
      301 gaaacaacgc ggcgagcttt gatcaacgac cttttggaaa cttcggcttc ccctggagag
      361 agcgagattc tccgcgctgt agaagtcacc attgttgtgc acgacgacat cattccgtgg
      421 cgttatccag ctaagcgcga actgcaattt ggagaatggc agcgcaatga cattcttgca
      481 ggtatcttcg agccagccac gatcgacatt gatctggcta tcttgctgac aaaagcaaga
      541 gaacatagcg ttgccttggt aggtccagcg gcggaggaac tctttgatcc ggttcctgaa
      601 caggatctat ttgaggcgct aaatgaaacc ttaacgctat ggaactcgcc gcccgactgg
      661 gctggcgatg agcgaaatgt agtgcttacg ttgtcccgca tttggtacag cgcagtaacc
      721 ggcaaaatcg cgccgaagga tgtcgctgcc gactgggcaa tggagcgcct gccggcccag
      781 tatcagcccg tcatacttga agctagacag gcttatcttg gacaagaaga agatcgcttg
      841 gcctcgcgcg cagatcagtt ggaagaattt gtccactacg tgaaaggcga gatcaccaag
      901 gtagtcggca aataagatgc cgctcgccag tcgattggct gagctcatga agttcctatt
      961 ccgaagttcc gcgaacgcgt aaaggatcta ggtgaagatc ctttttgata atctcatgac
     1021 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     1081 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     1141 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     1201 aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg
     1261 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     1321 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     1381 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     1441 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     1501 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     1561 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     1621 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     1681 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     1741 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
     1801 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
     1861 gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcatatgctg
     1921 gatccttgac agctagctca gtcctaggta taatactagt ctatactttc tagagaatgt
     1981 tttagagcta gaaatagcaa gttaaaataa ggctagtccg ttatcaactt gaaaaagtgg
     2041 caccgagtcg gtgctttttt tgaattctct agagtcgacc tgcagaagct tagatctatt
     2101 accctgttat ccctac
//
