LOCUS       sequence-272951-        6658 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MADR donor plasmid for FlpO+Cre- mediated insertion of a SM_FP-Myc, 
            a cytoplasmic fluorescent protein which includes multiple Myc 
            epitope tags.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6658)
  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 6658)
  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-Myc (dark) WPRE
FEATURES             Location/Qualifiers
     source          1..6658
                     /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)."
     regulatory      1388..1397
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1397..1426
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1430..1459
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1463..1492
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2024..2053
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2060..2089
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2102..2131
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2138..2167
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2381..2410
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2414..2443
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2447..2476
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     intron          2525..2657
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     2640..2659
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    2666..3254
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2719..2739)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3137..3148)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3281..3298)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3287..3398
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3494..3514)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3494..3512)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3533..3549)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3651..3670)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3716..3771
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3770..3789)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     protein_bind    4109..4142
                     /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(4161..4177)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4185..4201
                     /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(4209..4215,4216..4233,4234..4239))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4254..4275
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4334..4529
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4380..4515
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4535..4669
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4737..4754)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4908..5496)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4988..5007)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5667..6458,6459..6527))
                     /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     6290..6309
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6528..6632)
                     /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 accatggagc aaaaactgat atcggaggaa gacttggcag agcagaaact
     1441 gatttcggaa gaggatcttg ccgaacaaaa actgatcagt gaggaagacc ttgccggagt
     1501 gtcgaagggt gaagagctgt tcaccggcgt ggtgcccatc ctcgtcgagc tggacggaga
     1561 tgtcaatgga cataaattct ccgtacgggg agagggcgag ggcgacgcaa ccaacggtaa
     1621 actgaccctg aagttcattt gcaccacggg taaactgccc gtaccctggc ccaccctggt
     1681 cacaaccttg ggtggcggag ttcagtgttt ttcgcggtat ccagatcaca tgaagcagca
     1741 tgactttttc aagagcgcta tgcccgaggg ttacgttcaa gagaggacca tcagtttcaa
     1801 ggacgacggc acctacaaga cacgggctga ggtgaaattc gagggtgata cactcgtgaa
     1861 tcgtatcgag ctgaagggca tagattttaa ggaggatggc aatatcctgg gccacaagtt
     1921 ggagtataac ttcaacagtc acaatgtcta tataactgcc gataagcaga agaatggcat
     1981 taaagccaat ttcaaaattc gtcacaatgt cgaaggcggc gcagaacaga agctcatcag
     2041 tgaagaggat ctggccgcag agcagaaact cattagcgag gaggacttgg gcggaggcgg
     2101 cgagcagaag ctgatatcgg aggaagacct ggcggccgag caaaagctca tctcggagga
     2161 ggatctcgcg ggaggagatg gaagtgtcca gttggcggat cactaccagc agaatacgcc
     2221 aattggtgac ggccccgtcc ttctgccgga taaccactac ctctccaccc agtcggtgct
     2281 gagcaaggac ccgaatgaaa aacgtgacca catggttctt cttgagttcg tcacggctgc
     2341 gggcatcacg ttgggcatgg acgaactgta caagggcgct gaacagaaac ttatctccga
     2401 ggaggatctg gcggaacaga agctgatcag cgaagaggac ctggcggagc agaaacttat
     2461 cagcgaagag gatctgtaaa ccggtgctcg aggtaccgat gaatagctaa ggtcgaggcc
     2521 gcaggtaagt atcaaggtta caagacaggt ttaaggagac caatagaaac tgggcttgtc
     2581 gagacagaga agactcttgc gtttctgata ggcacctatt ggtcttactg acatccactt
     2641 tgcctttctc tccacaggtg tcgacaatca acctctggat tacaaaattt gtgaaagatt
     2701 gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc
     2761 tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg
     2821 gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac
     2881 tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc
     2941 cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc
     3001 ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa
     3061 gctgacgtcc tttccatggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc
     3121 cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc
     3181 ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg
     3241 ggccgcctcc ccgcctggaa ttcgagctcg gtacgatcag cctcgactgt gccttctagt
     3301 tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact
     3361 cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat
     3421 tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc
     3481 ccagcttttg ttccctttag tgagggttaa ttgcgcgctt ggcgtaatca tggtcatagc
     3541 tgtttcctgt gtgaaattgt tatccgctaa ttcactcctc aggtgcaggc tgcctatcag
     3601 aaggtggtgg ctggtgtggc caatgccctg gctcacaaat accactgaga tctttttccc
     3661 tctgccaaaa attatgggga catcatgaag ccccttgagc atctgacttc tggctaataa
     3721 aggaaattta ttttcattgc aatagtgtgt tggaattttt tgtgtctctc actcggaagg
     3781 acatatggga gggcaaatca tttaaaacat cagaatgagt atttggttta gagtttggca
     3841 acatatgcca tatgctggct gccatgaaca aaggtggcta taaagaggtc atcagtatat
     3901 gaaacagccc cctgctgtcc attccttatt ccatagaaaa gccttgactt gaggttagat
     3961 tttttttata ttttgttttg tgttattttt ttctttaaca tccctaaaat tttccttaca
     4021 tgttttacta gccagatttt tcctcctctc ctgactactc ccagtcatag ctgtccctct
     4081 tctcttatga agatccctcg acctgcagga agttcctatt ctctagaaag tataggaact
     4141 tcaagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     4201 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     4261 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     4321 tgccagcgga tccgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     4381 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     4441 ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag
     4501 gcttttttgg aggcctaggc ttttgcaaaa agctaacttg tttattgcag cttataatgg
     4561 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     4621 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggatcc gctgcattaa
     4681 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     4741 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     4801 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     4861 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     4921 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     4981 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     5041 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     5101 caatgctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     5161 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     5221 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     5281 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     5341 actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     5401 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     5461 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     5521 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     5581 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     5641 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     5701 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     5761 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     5821 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     5881 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     5941 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     6001 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     6061 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     6121 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     6181 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     6241 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     6301 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     6361 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     6421 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     6481 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     6541 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     6601 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctg
//
