LOCUS       sequence-172602-        7630 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  attB FlpStop plasmid, for insertion into MiMIC site .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7630)
  AUTHORS   Fisher YE, Yang HH, Isaacman-Beck J, Xie M, Gohl DM, Clandinin TR
  TITLE     FlpStop, a tool for conditional gene control in Drosophila.
  JOURNAL   Elife. 2017 Feb 17;6. pii: e22279. doi: 10.7554/eLife.22279.
  PUBMED    28211790
REFERENCE   2  (bases 1 to 7630)
  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: "Elife."; 
            date: "2017-02-17"; volume: "6. pii: e22279. doi"; pages: " 
            10.7554/eLife.22279"
COMMENT     Sequence Label: pFlpStop-attB-UAS-2.1-tdTom
FEATURES             Location/Qualifiers
     source          1..7630
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          1527..1592
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             1722..1742
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    2014..2148
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     intron          3157..3222
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             3352..3372
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    3644..3778
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    complement(3799..3832)
                     /label=FRT3
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision with other FRT3 sites
                     (Bosch et al., 2015)"
     protein_bind    complement(3897..3944)
                     /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)."
     promoter        complement(3974..4212)
                     /label=hsp70 promoter
                     /note="Drosophila melanogaster hsp70Bb promoter"
     primer_bind     complement(4006..4027)
                     /label=pCasper-hs
                     /note="Drosophila Hsp70 promoter, forward primer"
     protein_bind    4231..4325
                     /label=5X UAS
                     /bound_moiety="GAL4"
                     /note="five tandem copies of the ""ScaI site"" 17-mer 
                     CGGAGTACTGTCCTCCG, an upstream activating sequence (UAS) 
                     that efficiently binds yeast Gal4 (Webster et al., 1988; 
                     Pfeiffer et al., 2010)"
     protein_bind    4380..4449
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attB site for the phi-C31 integrase (Groth et al., 
                     2000)"
     primer_bind     complement(4489..4509)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4489..4507)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4528..4544)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4528..4544)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4541..4563)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4552..4568
                     /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(4576..4582,4583..4600,4601..4606))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4621..4642
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4759..4776)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4930..5518)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5010..5029)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5689..6480,6481..6549))
                     /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     6312..6331
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6550..6654)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6680..7135)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6817..6838)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7029..7048
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7262..7284
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7276..7293
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7277..7293
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7303..7322
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        7303..7321
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    7357..7426
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attB site for the phi-C31 integrase (Groth et al., 
                     2000)"
     protein_bind    7497..7530
                     /label=FRT3
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision with other FRT3 sites
                     (Bosch et al., 2015)"
     protein_bind    7581..7628
                     /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)."
ORIGIN
        1 ggccgctatg gtgtccaagg gcgaggaagt gatcaaggag ttcatgcgct tcaaagtgcg
       61 catggagggc agcatgaacg gccacgagtt cgagattgag ggcgagggcg agggacgccc
      121 atatgaggga acccagaccg ccaagctgaa agtgaccaag ggcggacccc tgcccttcgc
      181 ctgggatatt ctgagccccc agtttatgta cggcagcaag gcctacgtga agcaccccgc
      241 cgatatcccc gattacaaga agctgagctt ccccgagggc ttcaagtggg agcgcgtgat
      301 gaacttcgag gatggcggcc tcgtgaccgt gacccaggat agcagtctgc aggatggcac
      361 cctgatctac aaagtgaaga tgcgcggcac caacttcccc cccgatggac ccgtgatgca
      421 gaagaagacc atgggctggg aggccagcac cgagcgcctg tatccacgcg acggcgtgct
      481 gaagggcgag attcatcagg ccctgaagct gaaggacggc ggccactacc tggtggagtt
      541 caagaccatc tacatggcca agaagcccgt gcagctgccc ggctactact acgtggacac
      601 caagctggat atcaccagcc acaacgagga ttacaccatc gtggagcagt acgagcgcag
      661 cgagggccgc caccatctgt ttctgggaca cggaaccggc agcaccggaa gcggaagtag
      721 tggaaccgcc agcagcgagg ataacaacat ggccgtgatt aaggagttta tgcgctttaa
      781 agtgcgcatg gagggatcca tgaatggaca tgagtttgag atcgagggcg agggcgaggg
      841 acgcccctac gagggcacgc agacggccaa gctgaaagtg acgaagggcg gaccgctgcc
      901 gtttgcctgg gacatcctgt cgccgcagtt tatgtatggc tcgaaggcct atgtgaagca
      961 cccggccgac attccggact ataagaagct gtcgttccca gagggattca agtgggagcg
     1021 cgtgatgaat tttgaggacg gcggactcgt gaccgtgacg caggacagtt ccctgcagga
     1081 cggaacgctg atctataaag tgaagatgcg cggaacgaat ttcccgccgg acggcccagt
     1141 gatgcagaag aagacgatgg gatgggaggc ctcgaccgag cgcctgtacc cccgcgacgg
     1201 cgtgctgaag ggcgagatcc accaggccct gaagctgaag gatggcggac gctatctggt
     1261 ggagtttaag acgatctata tggccaagaa gccagtgcag ctgccgggat attactatgt
     1321 ggatacgaag ctggacatta cctcgcacaa tgaggactat acgattgtgg agcagtatga
     1381 gcgctcggag ggacgccatc acctgtttct gtacggcatg gatgagctgt ataagtagga
     1441 ggccggcctc gatctttgtg aaggaacctt acttctgtgg tgtgacataa ttggacaaac
     1501 tacctacaga gatttaaagc tctaaggtaa atataaaatt tttaagtgta taatgtgtta
     1561 aactactgat tctaattgtt tgtgtatttt agattccaac ctatggaact gatgaatggg
     1621 agcagtggtg gaatgccttt aatgaggaaa acctgttttg ctcagaagaa atgccatcta
     1681 gtgatgatga ggctactgct gactctcaac attctactcc tccaaaaaag aagagaaagg
     1741 tagaagaccc caaggacttt ccttcagaat tgctaagttt tttgagtcat gctgtgttta
     1801 gtaatagaac tcttgcttgc tttgctattt acaccacaaa ggaaaaagct gcactgctat
     1861 acaagaaaat tatggaaaaa tatttgatgt atagtgcctt gactagagat cataatcagc
     1921 cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac
     1981 ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt
     2041 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     2101 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggatctg cgtcgcgcca
     2161 cttcaacgct cgatgggagc gtcattggtg ggcggggtaa ccgtcgaaat cagtgtttac
     2221 gcttccaatc gcaacaaaaa attcactgca acactgaaaa gcatacgaaa acgatgaaga
     2281 ttgtacgaga aaccataaag tattttatcc acaaagacac gtatagcaga aaagccaagt
     2341 taactcggcg ataagttgtg tacacaagaa taaaatcggc cagattcagt gttgtcagaa
     2401 ataagaaaac cccactatgt ttttctttgc cttttctttc tcccagcgat cattcatttc
     2461 gtggtgaaag aacggggtca ttgcacggag tttcgactgc gggaaagcag agctgccgtt
     2521 cacttcgtct ataattagcg ctttctattt tccccgattc gggccgctgc tgcgcttttc
     2581 cgcctgctgt ttgtggcaag tgtagcagca ggctgtgcac gcagtgtggc atgcacttgg
     2641 ctttccaccg ttggtatcga ttctctggga cgatgagtca ttcctttcgg ggccacagca
     2701 taatcgttgc cagctcaccg aaatggtgac ttcatttctt aactgccgtc aagcatgcga
     2761 ttgtacatac atacatattt atatatgtac atatttatgt gactatggta ggtcgatata
     2821 atagcaatca acgcaagcaa atgtgtcagt cctgcttaca ggaacgattc tatttagtaa
     2881 ttttcgttgt ataaagtaat tatgtatgta tgtaagcccc ataaatctga aaaccggtac
     2941 tagtacgaaa gtcgatccaa catggcgact tgtcccatcc ccggcatgtt taaatatact
     3001 aattattctt gaactaattt taatcaaccg atttatctct cttccgcagg tctaactaac
     3061 taacaaaatt gactaagtga gatctttgtg aaggaacctt acttctgtgg tgtgacataa
     3121 ttggacaaac tacctacaga gatttaaagc tctaaggtaa atataaaatt tttaagtgta
     3181 taatgtgtta aactactgat tctaattgtt tgtgtatttt agattccaac ctatggaact
     3241 gatgaatggg agcagtggtg gaatgccttt aatgaggaaa acctgttttg ctcagaagaa
     3301 atgccatcta gtgatgatga ggctactgct gactctcaac attctactcc tccaaaaaag
     3361 aagagaaagg tagaagaccc caaggacttt ccttcagaat tgctaagttt tttgagtcat
     3421 gctgtgttta gtaatagaac tcttgcttgc tttgctattt acaccacaaa ggaaaaagct
     3481 gcactgctat acaagaaaat tatggaaaaa tatttgatgt atagtgcctt gactagagat
     3541 cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct
     3601 ccccctgaac tcgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc
     3661 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     3721 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggatctt
     3781 aattaaatcg atcctaggga agttcctata ctatttgaag aataggaact tcggaatagg
     3841 aacttctaag aggtttcata ttgctaatag cagctacaat ccagctacca ttctgcgaag
     3901 ttcctatact ttctagagaa taggaacttc ggaataggaa cttcgctggt accctcgagc
     3961 ccaattccct attcagagtt ctcttcttgt attcaataat tacttcttgg cagatttcag
     4021 tagttgcagt tgatttactt ggttgctggt tacttttaat tgattcactt taacttgcac
     4081 tttactgcag attgtttagc ttgttcagct gcgcttgttt atttgcttag ctttcgctta
     4141 gcgacgtgtt cactttgctt gtttgaattg aattgtcgct ccgtagacga agcgcctcta
     4201 tttatactcc ggcgctcgct agagtctccg ctcggaggac agtactccgc tcggaggaca
     4261 gtactccgct cggaggacag tactccgctc ggaggacagt actccgctcg gaggacagta
     4321 ctccgacctg cagggaattc ctgcagacta gttctagagt cgaccatcat gatggaccag
     4381 atgggtgagg tggagtacgc gcccggggag cccaagggca cgccctggca cccgcaccgc
     4441 ggcttcgaga ccgtgaccta catcatgcat ggtacccagc ttttgttccc tttagtgagg
     4501 gttaattgcg cgcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     4561 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     4621 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     4681 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     4741 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     4801 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     4861 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     4921 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     4981 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     5041 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     5101 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     5161 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     5221 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     5281 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     5341 gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa
     5401 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     5461 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     5521 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     5581 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     5641 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     5701 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     5761 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     5821 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     5881 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     5941 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     6001 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     6061 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     6121 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     6181 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     6241 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     6301 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     6361 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     6421 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     6481 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     6541 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     6601 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     6661 tccccgaaaa gtgccaccta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat
     6721 ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa
     6781 tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta
     6841 ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca
     6901 ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat
     6961 cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg
     7021 agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc
     7081 acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcccat
     7141 tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta
     7201 cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt
     7261 tcccagtcac gacgttgtaa aacgacggcc agtgagcgcg cgtaatacga ctcactatag
     7321 ggcgaattgg agctcgctag cgatgtaggt cacggtctcg aagccgcggt gcgggtgcca
     7381 gggcgtgccc ttgggctccc cgggcgcgta ctccacctca cccatctggt ccatcatgat
     7441 ggtcgacaag cttggatccg atgtcccaat acgcgtaggc ctgaagttcc tattccgaag
     7501 ttcctattct tcaaatagta taggaacttc tctttgcacc atttcaaaga ataacagtga
     7561 taatttctgg gttaaggcaa gaagttccta ttccgaagtt cctattctct agaaagtata
     7621 ggaacttcgc
//
