LOCUS       sequence-272954-        6621 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 6621)
  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 6621)
  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..6621
                     /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..3217
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2683..2703)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3100..3111)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3244..3261)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3250..3361
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3457..3477)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3457..3475)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3496..3512)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3614..3633)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3679..3734
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3733..3752)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     protein_bind    4072..4105
                     /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(4124..4140)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4148..4164
                     /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(4172..4178,4179..4196,4197..4202))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4217..4238
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4297..4492
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4343..4478
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4498..4632
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4700..4717)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4871..5459)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4951..4970)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5630..6421,6422..6490))
                     /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     6253..6272
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6491..6595)
                     /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 cantccgtgg tgttgtcggg gaagctgacg tcctttccat ggctgctcgc ctgtgttgcc
     3061 acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac
     3121 cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct
     3181 cagacgagtc ggatctccct ttgggccgcc tccccgcctg gaattcgagc tcggtacgat
     3241 cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt
     3301 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat
     3361 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg
     3421 gggaggattg ggaagacaat agcccagctt ttgttccctt tagtgagggt taattgcgcg
     3481 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc taattcactc
     3541 ctcaggtgca ggctgcctat cagaaggtgg tggctggtgt ggccaatgcc ctggctcaca
     3601 aataccactg agatcttttt ccctctgcca aaaattatgg ggacatcatg aagccccttg
     3661 agcatctgac ttctggctaa taaaggaaat ttattttcat tgcaatagtg tgttggaatt
     3721 ttttgtgtct ctcactcgga aggacatatg ggagggcaaa tcatttaaaa catcagaatg
     3781 agtatttggt ttagagtttg gcaacatatg ccatatgctg gctgccatga acaaaggtgg
     3841 ctataaagag gtcatcagta tatgaaacag ccccctgctg tccattcctt attccataga
     3901 aaagccttga cttgaggtta gatttttttt atattttgtt ttgtgttatt tttttcttta
     3961 acatccctaa aattttcctt acatgtttta ctagccagat ttttcctcct ctcctgacta
     4021 ctcccagtca tagctgtccc tcttctctta tgaagatccc tcgacctgca ggaagttcct
     4081 attctctaga aagtatagga acttcaagct tggcgtaatc atggtcatag ctgtttcctg
     4141 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     4201 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     4261 ctttccagtc gggaaacctg tcgtgccagc ggatccgcat ctcaattagt cagcaaccat
     4321 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     4381 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga
     4441 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctaac
     4501 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     4561 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
     4621 catgtctgga tccgctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     4681 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     4741 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     4801 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     4861 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     4921 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     4981 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     5041 cttcgggaag cgtggcgctt tctcaatgct cacgctgtag gtatctcagt tcggtgtagg
     5101 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     5161 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     5221 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     5281 agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga
     5341 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     5401 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     5461 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     5521 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     5581 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     5641 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     5701 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     5761 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     5821 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     5881 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     5941 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     6001 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     6061 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     6121 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     6181 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     6241 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     6301 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     6361 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     6421 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     6481 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     6541 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     6601 ttccccgaaa agtgccacct g
//
