LOCUS       sequence-222447-        2431 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 2431)
  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 2431)
  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: Control-cx43.4-48H-pGTag
FEATURES             Location/Qualifiers
     source          1..2431
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     complement(56..75)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(57..75)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(80..96)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(80..96)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(198..217)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             209..1018
                     /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     complement(276..295)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      1165..1753
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1654..1673
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1907..1924
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2083..2110)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(2202..2288)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     2337..2359
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2351..2368
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2352..2368
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 aaaccgctgt ggcccgaacg cctcccgcta gcgatgggat ctcggccgcg atatccccta
       61 tagtgagtcg tattacatgg tcatagctgt ttcctggcag ctctggcccg tgtctcaaaa
      121 tctctgatgt tacattgcac aagataaaaa tatatcatca tgaacaataa aactgtctgc
      181 ttacataaac agtaatacaa ggggtgttat gagccatatt caacgggaaa cgtcgaggcc
      241 gcgattaaat tccaacatgg atgctgattt atatgggtat aaatgggctc gcgataatgt
      301 cgggcaatca ggtgcgacaa tctatcgctt gtatgggaag cccgatgcgc cagagttgtt
      361 tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg tcagactaaa
      421 ctggctgacg gaatttatgc ctcttccgac catcaagcat tttatccgta ctcctgatga
      481 tgcatggtta ctcaccactg cgatccccgg aaaaacagca ttccaggtat tagaagaata
      541 tcctgattca ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc ggttgcattc
      601 gattcctgtt tgtaattgtc cttttaacag cgatcgcgta tttcgtctcg ctcaggcgca
      661 atcacgaatg aataacggtt tggttgatgc gagtgatttt gatgacgagc gtaatggctg
      721 gcctgttgaa caagtctgga aagaaatgca taaacttttg ccattctcac cggattcagt
      781 cgtcactcat ggtgatttct cacttgataa ccttattttt gacgagggga aattaatagg
      841 ttgtattgat gttggacgag tcggaatcgc agaccgatac caggatcttg ccatcctatg
      901 gaactgcctc ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa aatatggtat
      961 tgataatcct gatatgaata aattgcagtt tcatttgatg ctcgatgagt ttttctaatc
     1021 agaattggtt aattggttgt aacactggca gagcattacg ctgacttgac gggacggcgc
     1081 aagctcatga ccaaaatccc ttaacgtgag ttacgcgtcg ttccactgag cgtcagaccc
     1141 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     1201 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     1261 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt
     1321 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     1381 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     1441 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     1501 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagcattg
     1561 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     1621 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     1681 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     1741 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     1801 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     1861 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     1921 cgaggaagcg gatgtgcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca
     1981 ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat
     2041 taatacgcgt accgctagcc aggaagagtt tgtagaaacg caaaaaggcc atccgtcagg
     2101 atggccttct gcttagtttg atgcctggca gtttatggcg ggcgtcctgc ccgccaccct
     2161 ccgggccgtt gcttcacaac gttcaaatcc gctcccggcg gatttgtcct actcaggaga
     2221 gcgttcaccg acaaacaaca gataaaacga aaggcccagt cttccgactg agcctttcgt
     2281 tttatttgat gcctggcagt tccctactct cgcgttaacg ctagcatgga tgttttccca
     2341 gtcacgacgt tgtaaaacga cggccagtct taagctcggg ccctgcagct ctagagctcg
     2401 aattcgggag gcgttcgggc cacagcggcc c
//
