LOCUS       sequence-103225-        3474 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor vector for iRFP knock-in via landing pad.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3474)
  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 3474)
  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: pBGT
FEATURES             Location/Qualifiers
     source          1..3474
                     /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      293..302
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    1373..1494
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1410..1429)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1464..1483
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(1610..1657)
                     /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(1658..1691)
                     /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(1775..1791)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1775..1791)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     terminator      1839..1870
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        1871..1973
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             1974..2789
                     /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(2047..2066)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      2813..2840
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(2852..3439)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2932..2951)
                     /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 tcgccaccat
      301 ggccgaaggg agcgtggcaa gacagcccga tctgctgaca tgcgacgacg aaccaatcca
      361 cattccagga gctatccagc ctcatggact gctgctggct ctggccgctg acatgactat
      421 cgtggcaggc tctgataacc tgccagagct gaccgggctg gctatcggag cactgattgg
      481 ccggtccgca gccgacgtgt tcgattctga gactcacaat agactgacca tcgccctggc
      541 tgaacctggg gctgcagtgg gagccccaat taccgtcggc ttcacaatgc gaaaggacgc
      601 tggctttatc gggtcctggc accgccatga tcagctgatt tttctggagc tggaaccacc
      661 tcagagagac gtggcagagc cacaggcttt ctttcggaga acaaacagcg ccatcaggcg
      721 cctgcaggcc gctgagactc tggaatccgc ttgcgcagcc gctgcacagg aagtgaggaa
      781 gatcaccggg ttcgatcgcg tcatgatcta cagattcgcc tcagacttta gcggagaagt
      841 gatcgcagaa gatcgctgtg ccgaggtcga aagtaaactg ggcctgcatt acccagcctc
      901 aaccgtgcct gcacaggcac gacggctgta tacaatcaac cccgtgcgga tcattcccga
      961 cattaattac agaccagtgc ccgtcacacc tgacctgaac ccagtgactg ggaggcccat
     1021 cgatctgagc ttcgccattc tgcgcagcgt gtcccctgtc cacctggagt ttatgaggaa
     1081 tatcggaatg catggcacca tgtctatcag tattctgaga ggcgaaaggc tgtgggggct
     1141 gatcgtgtgc caccatcgga caccttacta tgtggacctg gacggcaggc aggcttgtga
     1201 actggtggct caggtgctgg cttggcagat tggcgtgatg gaagaataag gccggccgcg
     1261 actctagatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct
     1321 cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt
     1381 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     1441 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttaaggcgt
     1501 aaattgtaag cgttaatgat ccactagagg atccccgggt accgagctcg aattcgaagt
     1561 tcctatactc ctataagaat aggaacttcg gaataggaac ttcatgatgg aagttcatat
     1621 actttctaga gaataggaac ttcggaatag gaacttcata acttcgtata ggatacttta
     1681 tacgaagtta tcagctgccg ctcgaggcgc gtgatcttac ggcattatac gtatgatcgg
     1741 tccacgatca gctagattat ctagtcagct tgatgtcata gctgtttcct gaggctcaat
     1801 actgaccatt taaatcatac ctgacctcca tagcagaaag tcaaaagcct ccgaccggag
     1861 gcttttgact tgatcggcac gtaagaggtt ccaactttca ccataatgaa ataagatcac
     1921 taccgggcgt attttttgag ttatcgagat tttcaggagc taaggaagct aaaatgagcc
     1981 atattcaacg ggaaacgtct tgcttgaagc cgcgattaaa ttccaacatg gatgctgatt
     2041 tatatgggta taaatgggct cgcgataatg tcgggcaatc aggtgcgaca atctatcgat
     2101 tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca
     2161 atgatgttac agatgagatg gtcaggctaa actggctgac ggaatttatg cctcttccga
     2221 ccatcaagca ttttatccgt actcctgatg atgcatggtt actcaccact gcgatcccag
     2281 ggaaaacagc attccaggta ttagaagaat atcctgattc aggtgaaaat attgttgatg
     2341 cgctggcagt gttcctgcgc cggttgcatt cgattcctgt ttgtaattgt ccttttaacg
     2401 gcgatcgcgt atttcgtctc gctcaggcgc aatcacgaat gaataacggt ttggttggtg
     2461 cgagtgattt tgatgacgag cgtaatggct ggcctgttga acaagtctgg aaagaaatgc
     2521 ataaactctt gccattctca ccggattcag tcgtcactca tggtgatttc tcacttgata
     2581 accttatttt tgacgagggg aaattaatag gttgtattga tgttggacga gtcggaatcg
     2641 cagaccgata ccaggatctt gccatcctat ggaactgcct cggtgagttt tctccttcat
     2701 tacagaaacg gctttttcaa aaatatggta ttgataatcc tgatatgaat aaattgcagt
     2761 ttcacttgat gctcgatgag tttttctaat gaggacctaa atgtaatcac ctggctcacc
     2821 ttcgggtggg cctttctgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     2881 atcacaaaaa tcgatgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     2941 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     3001 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     3061 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     3121 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     3181 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     3241 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     3301 ttggtatctg cgctctgctg aagccagtta cctcggaaaa agagttggta gctcttgatc
     3361 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     3421 cagaaaaaaa ggatctcaag aagatccttt gattttctac cgaagaaagg ccca
//
