LOCUS       sequence-305185-        6624 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MADR donor plasmid for FlpO+Cre- mediated insertion of SM_FP-HA, a 
            cytoplasmic fluorescent protein which includes multiple HA epitope 
            tags.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6624)
  AUTHORS   Kim GB, Rincon Fernandez Pacheco D, Saxon D, Yang A, Sabet S, 
            Dutra-Clarke M, Levy R, Watkins A, Park H, Abbasi Akhtar A, Linesch 
            PW, Kobritz N, Chandra SS, Grausam K, Ayala-Sarmiento A, Molina J, 
            Sedivakova K, Hoang K, Tsyporin J, Gareau DS, Filbin MG, Bannykh S, 
            Santiskulvong C, Wang Y, Tang J, Suva ML, Chen B, Danielpour M, 
            Breunig JJ
  TITLE     Rapid Generation of Somatic Mouse Mosaics with Locus-Specific, 
            Stably Integrated Transgenic Elements.
  JOURNAL   Cell. 2019 Sep 19;179(1):251-267.e24. doi: 
            10.1016/j.cell.2019.08.013.
  PUBMED    31539496
REFERENCE   2  (bases 1 to 6624)
  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"; doi: 
            "10.1016/j.cell.2019.08"; journalName: "Cell"; date: "2019-09-19- 
            19"; volume: "179"; issue: "1"; pages: "251-267.e24"
COMMENT     Sequence Label: MADR pDonor smFP-HA (dark) WPRE
FEATURES             Location/Qualifiers
     source          1..6624
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    32..493
                     /label=PGK poly(A) signal
                     /note="mouse phosphoglycerate kinase 1 polyadenylation 
                     signal"
     polyA_signal    641..775
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    895..1029
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    1149..1283
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    complement(1312..1345)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             1397..1423
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1427..1453
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1457..1483
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2012..2038
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2045..2071
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2084..2110
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2117..2143
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2354..2380
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2384..2410
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2414..2440
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     intron          2489..2621
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     2604..2623
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    2630..3218
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2683..2703)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3101..3112)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3245..3262)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3251..3362
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3458..3478)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3458..3476)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3497..3513)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3615..3634)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3680..3735
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3734..3753)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     protein_bind    4075..4108
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     primer_bind     complement(4127..4143)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4151..4167
                     /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(4175..4181,4182..4199,4200..4205))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4220..4241
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4300..4495
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4346..4481
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4501..4635
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4703..4720)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4874..5462)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4954..4973)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5633..6424,6425..6493))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6256..6275
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6494..6598)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggtcgacatt gattattgac tagtggatcc gctgtaagtc tgcagaaatt gatgatctat
       61 taaacaataa agatgtccac taaaatggaa gtttttcctg tcatactttg ttaagaaggg
      121 tgagaacaga gtacctacat tttgaatgga aggattggag ctacgggggt gggggtgggg
      181 tgggattaga taaatgcctg ctctttactg aaggctcttt actattgctt tatgataatg
      241 tttcatagtt ggatatcata atttaaacaa gcaaaaccaa attaagggcc agctcattcc
      301 tcccactcat gatctataga tctatagatc tctcgtggga tcattgtttt tctcttgatt
      361 cccactttgt ggttctaagt actgtggttt ccaaatgtgt cagtttcata gcctgaagaa
      421 cgagatcagc agcctctgtt ccacatacac ttcattctca gtattgtttt gccaagttct
      481 aattccatca gaagcttgca gatctgcgac tctagaggat ctgcgactct agaggatcat
      541 aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc
      601 cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta
      661 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
      721 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggatctgcga
      781 ctctagagga tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac
      841 ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg
      901 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
      961 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     1021 gtctggatct gcgactctag aggatcataa tcagccatac cacatttgta gaggttttac
     1081 ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg
     1141 ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     1201 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     1261 atgtatctta tcatgtctgg atccccatca agctgatccg gaacccttaa tataacttcg
     1321 tatagcatac attatacgaa gttattaggt ccctcgacct gcagcccaag ctagcgcaat
     1381 tgcggccgcc accatgtacc cttatgatgt gcccgattat gctggctacc cttatgatgt
     1441 tcctgattac gccggatatc cgtatgacgt gccagactac gcgggaggtg tgagcaaggg
     1501 agaggagttg tttaccggcg tggtccccat actggtggag ttggatggcg acgttaatgg
     1561 acataaattc tcggtccgcg gcgagggaga gggagacgcc accaatggca agctgacgct
     1621 taagttcatt tgcacgactg gaaaattgcc tgtcccctgg ccgaccctgg tcaccacact
     1681 gggcggagga gtgcagtgct tctcccgtta cccagaccac atgaagcagc acgatttctt
     1741 caagagcgca atgccggagg gatacgtgca agaacggact atttccttca aggatgacgg
     1801 aacctataag acccgtgcgg aggtcaaatt cgagggtgac accctggtga accgaattga
     1861 actcaaagga atcgatttca aggaggatgg aaatattctg ggtcacaagc tggaatacaa
     1921 cttcaacagc cataatgtgt acattacggc tgataagcag aagaacggca ttaaggccaa
     1981 tttcaagatc cgccacaacg ttgagggtgg atatccctac gacgtgcccg attatgcggg
     2041 cggatatccg tatgatgttc cagattatgc tggtggtgga ggctatccct atgatgtccc
     2101 cgactacgcc ggaggttacc catacgacgt gcccgactac gctggaggcg atggcagcgt
     2161 gcagctggca gatcattatc aacagaatac ccccataggt gatggccccg ttctgcttcc
     2221 agataatcac tacctttcca cccagagcgt gctttcgaaa gacccgaacg aaaagcgtga
     2281 tcacatggtc ctgctggagt ttgtgaccgc ggctggaatc accctgggta tggacgaact
     2341 ctacaagggt ggctacccct acgatgtgcc ggattacgct ggatatccgt atgacgtacc
     2401 ggactatgcc ggttacccgt acgatgtccc ggactacgct taaaccggtg ctcgaggtac
     2461 cgatgaatag ctaaggtcga ggccgcaggt aagtatcaag gttacaagac aggtttaagg
     2521 agaccaatag aaactgggct tgtcgagaca gagaagactc ttgcgtttct gataggcacc
     2581 tattggtctt actgacatcc actttgcctt tctctccaca ggtgtcgaca atcaacctct
     2641 ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct
     2701 atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat
     2761 tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt
     2821 caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat
     2881 tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc
     2941 ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga
     3001 caattccgtg gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg cctgtgttgc
     3061 cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga
     3121 ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc
     3181 tcagacgagt cggatctccc tttgggccgc ctccccgcct ggaattcgag ctcggtacga
     3241 tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct
     3301 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca
     3361 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag
     3421 ggggaggatt gggaagacaa tagcccagct tttgttccct ttagtgaggg ttaattgcgc
     3481 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctaattcact
     3541 cctcaggtgc aggctgccta tcagaaggtg gtggctggtg tggccaatgc cctggctcac
     3601 aaataccact gagatctttt tccctctgcc aaaaattatg gggacatcat gaagcccctt
     3661 gagcatctga cttctggcta ataaaggaaa tttattttca ttgcaatagt gtgttggaat
     3721 tttttgtgtc tctcactcgg aaggacatat gggagggcaa atcatttaaa acatcagaat
     3781 gagtatttgg tttagagttt ggcaacatat gcccatatgc tggctgccat gaacaaaggt
     3841 tggctataaa gaggtcatca gtatatgaaa cagccccctg ctgtccattc cttattccat
     3901 agaaaagcct tgacttgagg ttagattttt tttatatttt gttttgtgtt atttttttct
     3961 ttaacatccc taaaattttc cttacatgtt ttactagcca gatttttcct cctctcctga
     4021 ctactcccag tcatagctgt ccctcttctc ttatggagat ccctcgacct gcaggaagtt
     4081 cctattctct agaaagtata ggaacttcaa gcttggcgta atcatggtca tagctgtttc
     4141 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     4201 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     4261 ccgctttcca gtcgggaaac ctgtcgtgcc agcggatccg catctcaatt agtcagcaac
     4321 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     4381 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc
     4441 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct
     4501 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     4561 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     4621 tatcatgtct ggatccgctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     4681 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     4741 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     4801 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     4861 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     4921 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     4981 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     5041 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     5101 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     5161 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     5221 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     5281 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     5341 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     5401 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     5461 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     5521 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     5581 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     5641 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     5701 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     5761 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     5821 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     5881 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     5941 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     6001 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     6061 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     6121 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     6181 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     6241 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     6301 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     6361 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     6421 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     6481 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     6541 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     6601 catttccccg aaaagtgcca cctg
//
