LOCUS       sequence-276300-        5285 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cassette for testing FLPe recombinase activity using a fluorescent 
            reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5285)
  AUTHORS   Cody JP, Graham ND, Zhao C, Swyers NC, Birchler JA
  TITLE     Site-specific recombinase genome engineering toolkit in maize.
  JOURNAL   Plant Direct. 2020 Mar 9;4(3):e00209. doi: 10.1002/pld3.209. 
            eCollection 2020 Mar.
  PUBMED    32166212
REFERENCE   2  (bases 1 to 5285)
  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: "Plant 
            Direct."; date: "2020-03-9"; volume: "4(3):e00209. doi"; pages: " 
            10.1002/pld3.209. eCollection 2020 Mar"
COMMENT     Sequence Label: pFLPe-AntiDsRed
FEATURES             Location/Qualifiers
     source          1..5285
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(82..99)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(253..841)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(333..352)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1012..1803,1804..1872))
                     /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     1635..1654
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1873..1944)
                     /gene="bla"
                     /label=AmpR promoter
     terminator      complement(1967..2219)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     protein_bind    2234..2267
                     /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)"
     CDS             2268..2945
                     /codon_start=1
                     /product="improved tetrameric variant of DsRed fluorescent 
                     protein"
                     /label=DsRed2
                     /note="mammalian codon-optimized"
                     /translation="MASSENVITEFMRFKVRMEGTVNGHEFEIEGEGEGRPYEGHNTVK
                     LKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
                     ATVTQDSSLQDGCFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGETHK
                     ALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDAKLDITSHNEDYTIVEQYERTEGRHH
                     LFL"
     primer_bind     complement(2448..2468)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     2860..2883
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     protein_bind    complement(2946..2979)
                     /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)"
     promoter        complement(2987..4977)
                     /label=Ubi promoter
                     /note="maize polyubiquitin gene promoter"
     primer_bind     complement(5097..5117)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5097..5115)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5136..5152)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5136..5152)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5149..5171)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5160..5176
                     /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(5184..5190,5191..5208,5209..5214))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5229..5250
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
       61 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
      121 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
      181 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
      241 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
      301 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
      361 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
      421 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
      481 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
      541 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
      601 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
      661 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
      721 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
      781 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
      841 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
      901 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
      961 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     1021 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     1081 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     1141 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     1201 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     1261 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     1321 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     1381 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     1441 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     1501 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     1561 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     1621 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     1681 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     1741 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     1801 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     1861 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     1921 catgagcgga tacatatttg aatgtatagc ttgatatcga attcccgatc tagtaacata
     1981 gatgacaccg cgcgcgataa tttatcctag tttgcgcgct atattttgtt ttctatcgcg
     2041 tattaaatgt ataattgcgg gactctaatc ataaaaaccc atctcataaa taacgtcatg
     2101 cattacatgt taattattac atgcttaacg taattcaaca gaaattatat gataatcatc
     2161 gcaagaccgg caacaggatt caatcttaag aaactttatt gccaaatgtt tgaacgatcg
     2221 gggatccact agtgaagttc ctattctcta gaaagtatag gaacttcatg gcctcctccg
     2281 agaacgtcat caccgagttc atgcgcttca aggtgcgcat ggagggcacc gtgaacggcc
     2341 acgagttcga gatcgagggc gagggcgagg gccgccccta cgagggccac aacaccgtga
     2401 agctgaaggt gaccaagggc ggccccctgc ccttcgcctg ggacatcctg tccccccagt
     2461 tccagtacgg ctccaaggtg tacgtgaagc accccgccga catccccgac tacaagaagc
     2521 tgtccttccc cgagggcttc aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg
     2581 cgaccgtgac ccaggactcc tccctgcagg acggctgctt catctacaag gtgaagttca
     2641 tcggcgtgaa cttcccctcc gacggccccg tgatgcagaa gaagaccatg ggctgggagg
     2701 cctccaccga gcgcctgtac ccccgcgacg gcgtgctgaa gggcgagacc cacaaggccc
     2761 tgaagctgaa ggacggcggc cactacctgg tggagttcaa gtccatctac atggccaaga
     2821 agcccgtgca gctgcccggc tactactacg tggacgccaa gctggacatc acctcccaca
     2881 acgaggacta caccatcgtg gagcagtacg agcgcaccga gggccgccac cacctgttcc
     2941 tgtaggaagt tcctatactt tctagagaat aggaacttca tttaaatgca gaagtaacac
     3001 caaacaacag ggtgagcatc gacaaaagaa acagtaccaa gcaaataaat agcgtatgaa
     3061 ggcagggcta aaaaaatcca catatagctg ctgcatatgc catcatccaa gtatatcaag
     3121 atcaaaataa ttataaaaca tacttgttta ttataataga taggtactca aggttagagc
     3181 atatgaatag atgctgcata tgccatcatg tatatgcatc agtaaaaccc acatcaacat
     3241 gtatacctat cctagatcga tatttccatc catcttaaac tcgtaactat gaagatgtat
     3301 gacacacaca tacagttcca aaattaataa atacaccagg tagtttgaaa cagtattcta
     3361 ctccgatcta gaacgaatga acgaccgccc aaccacacca catcatcaca accaagcgaa
     3421 caaaaagcat ctctgtatat gcatcagtaa aacccgcatc aacatgtata cctatcctag
     3481 atcgatattt ccatccatca tcttcaattc gtaactatga atatgtatgg cacacacata
     3541 cagatccaaa attaataaat ccaccaggta gtttgaaaca gaattctact ccgatctaga
     3601 acgaccgccc aaccagacca catcatcaca accaagacaa aaaaaagcat gaaaagatga
     3661 cccgacaaac aagtgcacgg catatattga aataaaggaa aagggcaaac caaaccctat
     3721 gcaacgaaac aaaaaaaatc atgaaatcga tcccgtctgc ggaacggcta gagccatccc
     3781 aggattcccc aaagagaaac actggcaagt tagcaatcag aacgtgtctg acgtacaggt
     3841 cgcatccgtg tacgaacgct agcagcacgg atctaacaca aacacggatc taacacaaac
     3901 atgaacagaa gtagaactac cgggccctaa ccatggaccg gaacgccgat ctagagaagg
     3961 tagaganggg ggggggggga ggacgagcgg cgtaccttga agcggaggtg ccgacgggtg
     4021 gatttggggg agatctggtt gtgtgtgtgt gcgctccgaa caacacgagg ttggggaaag
     4081 agggtgtgga gggggtgtct atttattacg gcgggcgagg aagggaaagc gaaggagcgg
     4141 tgggaaagga atcccccgta gctgccggtg ccgtgagagg aggaggaggc cgcctgccgt
     4201 gccggctcac gtctgccgct ccgccacgca atttctggat gccgacagcg gagcaagtcc
     4261 aacggtggag cggaactctc gagaggggtc cagaggcagc gacagagatg ccgtgccgtc
     4321 tgcttcgctt ggcccgacgc gacgctgctg gttcgctggt tggtgtccgt tagactcgtc
     4381 gacggcgttt aacaggctgg cattatctac tcgaaacaag aaaaatgttt ccttagtttt
     4441 tttaatttct taaagggtat ttgtttaatt tttagtcact ttattttatt ctattttata
     4501 tctaaattat taaataaaaa aactaaaata gagttttagt tttcttaatt tagaggctaa
     4561 aatagaataa aatagatgta ctaaaaaaat tagtctataa aaaccattaa ccctaaaccc
     4621 taaatggatg tactaataaa atggatgaag tattatatag gtgaagctat ttgcaaaaaa
     4681 aaaggagaac acatgcacac taaaaagata aaactgtaga gtcctgttgt caaaatactc
     4741 aattgtcctt tagaccatgt ctaactgttc atttatatga ttctctaaaa cactgatatt
     4801 attgtagtac tatagattat attattcgta gagtaaagtt taaatatatg tataaagata
     4861 gataaactgc acttcaaaca agtgtgacaa aaaaaatatg tggtaatttt ttataactta
     4921 gacatgcaat gctcattatc tctagagagg ggcacgaccg ggtcacgctg cactgcatgc
     4981 tgcagcccgg gcgacgtcga ctgtttaaac gaggccttaa ttaagggccc aattggccac
     5041 gtacgtctag atctggcgcg cctcgaggat cgatccatgg tacccagctt ttgttccctt
     5101 tagtgagggt taattgcgcg cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     5161 tgttatccgc tcacaattcc acacaacata cgagccggga gcataaagtg taaagcctgg
     5221 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     5281 tcggg
//
