LOCUS       sequence-220868-        2115 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 2115)
  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 2115)
  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..2115
                     /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      1089..1677
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1578..1597
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1831..1848
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        1925..1959
                     /label=J23119(SpeI) promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119) modified to end with an SpeI site"
     misc_RNA        1978..2053
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
ORIGIN
        1 tcgagttcat gtgcagctcc ataagcaaaa 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 gagctcataa gttcctattc
      961 cgaagttccg cgaacgcgta aaggatctag gtgaagatcc tttttgataa tctcatgacc
     1021 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     1081 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     1141 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     1201 actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc
     1261 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     1321 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     1381 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     1441 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     1501 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     1561 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     1621 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     1681 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     1741 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     1801 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
     1861 cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg catatgctgg
     1921 atccttgaca gctagctcag tcctaggtat aatactagtc tatactttct agagaatgtt
     1981 ttagagctag aaatagcaag ttaaaataag gctagtccgt tatcaacttg aaaaagtggc
     2041 accgagtcgg tgcttttttt gaattctcta gagtcgacct gcagaagctt agatctatta
     2101 ccctgttatc cctac
//
