LOCUS       sequence-230149-        1062 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Donor for precise CRISPR directed genomic integration using the 
            GeneWeld method.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1062)
  AUTHORS   Wierson WA, Welker JM, Almeida MP, Mann CM, Webster DA, Torrie ME, 
            Weiss TJ, Kambakam S, Vollbrecht MK, Lan M, McKeighan KC, Levey J, 
            Ming Z, Wehmeier A, Mikelson CS, Haltom JA, Kwan KM, Chien CB, 
            Balciunas D, Ekker SC, Clark KJ, Webber BR, Moriarity BS, Solin SL, 
            Carlson DF, Dobbs DL, McGrail M, Essner J
  TITLE     Efficient targeted integration directed by short homology in 
            zebrafish and mammalian cells.
  JOURNAL   Elife. 2020 May 15;9. pii: 53968. doi: 10.7554/eLife.53968.
  PUBMED    32412410
REFERENCE   2  (bases 1 to 1062)
  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: "Elife."; 
            date: "2020-05-15"; volume: "9. pii: 53968. doi"; pages: " 
            10.7554/eLife.53968"
COMMENT     Sequence Label: pPRISM-Cre-cmlc2-TagBFP
FEATURES             Location/Qualifiers
     source          1..1062
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(46..459)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWTCHTSRSHELLS
                     VKASRRRGRKPRSYVQFTDRTPKFACITGRCNE"
     CDS             complement(108..164)
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     primer_bind     309..331
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     323..340
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     324..340
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     350..369
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        350..368
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(416..434)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(455..471)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(455..471)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(468..490)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    479..495
                     /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(503..509,510..527,528..533))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    548..569
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     misc_feature    join(656..818,819..830)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(860..877)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gcctggcgat ccctgaacat gtccatcagg ttcttgcgaa cctcatcact cgttgcatcg
       61 accggtaatg caggcaaatt ttggtgtacg gtcagtaaat tggacatagg accggggttt
      121 tcttccacgt ctcctgcttg ctttaacaga gagaagttcg tggctccggg aagtgtggca
      181 ggtccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc
      241 gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc
      301 agggttttcc cagtcacgac gttgtaaaac gacggccagt gagcgcgcgt aatacgactc
      361 actatagggc gaattgggta ccgggccccc cctcgaggtc ctccagcttt tgttcccttt
      421 agtgagggtt aattgcgcgc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
      481 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
      541 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg cacctgcgtg tccgctgtgg
      601 cccgaacgcc tcccatcaat tgaccctcac taaagggaga ccactctagt tttctttctt
      661 gcttttactt ttacttcctt aatactcaag tacaatttta atggagtact tttttacttt
      721 tactcaagta agattctagc cagatacttt tacttttaat tgagtaaaat tttccctaag
      781 tacttgtact ttcacttgag taaaattttt gagtactttt tacacctctg ctcggtacca
      841 tttaaatctg ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
      901 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
      961 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     1021 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tc
//
