LOCUS       sequence-103224-        3593 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor vector for Dre knock-in via landing pad.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3593)
  AUTHORS   Ohtsuka M, Miura H, Mochida K, Hirose M, Hasegawa A, Ogura A, 
            Mizutani R, Kimura M, Isotani A, Ikawa M, Sato M, Gurumurthy CB
  TITLE     One-step generation of multiple transgenic mouse lines using an 
            improved Pronuclear Injection-based Targeted Transgenesis (i-PITT).
  JOURNAL   BMC Genomics. 2015 Apr 9;16:274. doi: 10.1186/s12864-015-1432-5.
  PUBMED    25887549
REFERENCE   2  (bases 1 to 3593)
  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: "BMC 
            Genomics."; date: "2015-04-9"; volume: "16:274. doi"; pages: " 
            10.1186/s12864-015-1432-5"
COMMENT     Sequence Label: pBGX
FEATURES             Location/Qualifiers
     source          1..3593
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     5..22
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6..22
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    161..194
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     protein_bind    complement(203..236)
                     /label=loxJTZ17
                     /bound_moiety="Cre recombinase"
                     /note="Right element (RE) mutant of loxP (Araki et al., 
                     2010). Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     regulatory      292..301
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             307..1329
                     /product="site-specific recombinase"
                     /label=Dre recombinase
                     /note="Dre is a site-specific recombinase from 
                     bacteriophage D6. Recombination occurs at rox sequences."
     CDS             1345..1365
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    1492..1613
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1529..1548)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1583..1602
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(1729..1776)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     protein_bind    complement(1777..1810)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     primer_bind     complement(1894..1910)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1894..1910)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     terminator      1958..1989
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        1990..2092
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             2093..2908
                     /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="MSHIQRETSCLKPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNGDRVFRLAQAQSRMNNGLVGA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(2166..2185)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      2932..2959
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(2971..3558)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3051..3070)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cccgtgtaaa acgacggcca gtttatctag tcagcttgat tctagctgat cgtggaccgg
       61 aaggtgagcc agtgagttga ttgcagtcca gttacgctgg agtctgaggc tcgtcctgaa
      121 tgatatgcgg cctcggatcc gcgatcgatc cgcggtgcgg gtgccagggc gtgcccttgg
      181 gctccccggg cgcgtactcc acataacttc gtatagcata cattatagca atttatcgaa
      241 gttcctattc cgaagttcct attctatcag aagtatagga acttcaccgg tgccaccatg
      301 ggcgcctccg agctgatcat cagcggcagc agcggaggct tcctgagaaa catcggcaaa
      361 gagtaccagg aagccgccga gaacttcatg cggttcatga acgaccaggg cgcctacgcc
      421 cctaacaccc tgagggacct gagactggtg ttccacagct gggccagatg gtgccacgcc
      481 agacagctgg cctggttccc catcagcccc gagatggcca gagagtactt cctgcagctg
      541 cacgacgccg atctggccag caccaccatc gacaagcact acgccatgct gaacatgctg
      601 ctgagccact gcggcctgcc tccactgagc gacgacaaga gcgtgtccct ggccatgaga
      661 agaatcagaa gagaggccgc caccgagaag ggcgagagaa ccggccaggc catccccctg
      721 agatgggacg acctgaagct gctggacgtg ctgctgtcca gatccgagag actggtggac
      781 ctgaggaaca gagccttcct gttcgtggcc tacaacaccc tgatgaggat gagcgagatc
      841 agcagaatca gagtgggcga cctggaccag accggcgaca ccgtgaccct gcacatcagc
      901 cacaccaaga ccatcaccac agccgctggc ctggacaagg tgctgagcag acggaccacc
      961 gccgtgctga acgactggct ggacgtgtcc ggcctgagag aacaccccga cgccgtgctg
     1021 ttccccccaa tccacagaag caacaaggcc agaatcacca ccacccctct gaccgcccct
     1081 gccatggaaa agatcttcag cgacgcctgg gtgctgctga acaagaggga cgccaccccc
     1141 aacaagggca gatacagaac ctggaccggc cactccgcca gagtgggagc cgccatcgac
     1201 atggccgaga aacaggtgtc catggtggag atcatgcagg aaggcacctg gaagaaaccc
     1261 gagacactga tgagatacct gagaagaggc ggcgtgagcg tgggcgccaa cagcagactg
     1321 atggactccg ccagcggagc tggccccaag aaaaagcgga aagtgtgagg ccggccgcga
     1381 ctctagatca taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc
     1441 ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt
     1501 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     1561 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttaaggcgta
     1621 aattgtaagc gttaatgatc cactagagga tccccgggta ccgagctcga attcgaagtt
     1681 cctatactcc tataagaata ggaacttcgg aataggaact tcatgatgga agttcatata
     1741 ctttctagag aataggaact tcggaatagg aacttcataa cttcgtatag gatactttat
     1801 acgaagttat cagctgccgc tcgaggcgcg tgatcttacg gcattatacg tatgatcggt
     1861 ccacgatcag ctagattatc tagtcagctt gatgtcatag ctgtttcctg aggctcaata
     1921 ctgaccattt aaatcatacc tgacctccat agcagaaagt caaaagcctc cgaccggagg
     1981 cttttgactt gatcggcacg taagaggttc caactttcac cataatgaaa taagatcact
     2041 accgggcgta ttttttgagt tatcgagatt ttcaggagct aaggaagcta aaatgagcca
     2101 tattcaacgg gaaacgtctt gcttgaagcc gcgattaaat tccaacatgg atgctgattt
     2161 atatgggtat aaatgggctc gcgataatgt cgggcaatca ggtgcgacaa tctatcgatt
     2221 gtatgggaag cccgatgcgc cagagttgtt tctgaaacat ggcaaaggta gcgttgccaa
     2281 tgatgttaca gatgagatgg tcaggctaaa ctggctgacg gaatttatgc ctcttccgac
     2341 catcaagcat tttatccgta ctcctgatga tgcatggtta ctcaccactg cgatcccagg
     2401 gaaaacagca ttccaggtat tagaagaata tcctgattca ggtgaaaata ttgttgatgc
     2461 gctggcagtg ttcctgcgcc ggttgcattc gattcctgtt tgtaattgtc cttttaacgg
     2521 cgatcgcgta tttcgtctcg ctcaggcgca atcacgaatg aataacggtt tggttggtgc
     2581 gagtgatttt gatgacgagc gtaatggctg gcctgttgaa caagtctgga aagaaatgca
     2641 taaactcttg ccattctcac cggattcagt cgtcactcat ggtgatttct cacttgataa
     2701 ccttattttt gacgagggga aattaatagg ttgtattgat gttggacgag tcggaatcgc
     2761 agaccgatac caggatcttg ccatcctatg gaactgcctc ggtgagtttt ctccttcatt
     2821 acagaaacgg ctttttcaaa aatatggtat tgataatcct gatatgaata aattgcagtt
     2881 tcacttgatg ctcgatgagt ttttctaatg aggacctaaa tgtaatcacc tggctcacct
     2941 tcgggtgggc ctttctgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     3001 tcacaaaaat cgatgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     3061 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     3121 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     3181 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     3241 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     3301 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     3361 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     3421 tggtatctgc gctctgctga agccagttac ctcggaaaaa gagttggtag ctcttgatcc
     3481 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     3541 agaaaaaaag gatctcaaga agatcctttg attttctacc gaagaaaggc cca
//
