LOCUS       sequence-175676-        7986 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  FlpStop plasmid for CRISPR/HDR. Has MCS for homology arms.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7986)
  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 7986)
  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-HDR-UAS-2.1-tdTom
FEATURES             Location/Qualifiers
     source          1..7986
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    74..173
                     /label=phage phi-C31 attP
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attachment site of phage phi-C31"
     protein_bind    216..310
                     /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)"
     promoter        329..567
                     /label=hsp70 promoter
                     /note="Drosophila melanogaster hsp70Bb promoter"
     primer_bind     514..535
                     /label=pCasper-hs
                     /note="Drosophila Hsp70 promoter, forward primer"
     protein_bind    597..644
                     /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)."
     protein_bind    709..742
                     /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)"
     polyA_signal    763..897
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(1169..1189)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     intron          1319..1384
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     polyA_signal    2393..2527
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(2799..2819)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     intron          2949..3014
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     protein_bind    complement(4543..4590)
                     /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)."
     protein_bind    complement(4641..4674)
                     /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    4736..4769
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     regulatory      5004..5013
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5010..5687
                     /codon_start=1
                     /product="rapidly maturing tetrameric variant of DsRed 
                     fluorescent protein (Bevis and Glick, 2002)"
                     /label=DsRed-Express
                     /note="mammalian codon-optimized"
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGSFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGEIHK
                     ALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDSKLDITSHNEDYTIVEQYERAEGRHH
                     LFL"
     primer_bind     complement(5190..5210)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     5602..5625
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     polyA_signal    5808..5929
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    5945..5978
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    5980..6079
                     /label=phage phi-C31 attP
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attachment site of phage phi-C31"
     rep_origin      complement(6231..6819)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6311..6330)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6990..7781,7782..7850))
                     /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     7613..7632
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7851..7955)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ccaatgattc gctgaagcag gtggaattct tgcatgctag cggccgcgga catatgcaca
       61 cctgcgatcg tagcccaggt cagaagcggt tttcgggagt agtgccccaa ctggggtaac
      121 ctttgagttc tctcagttgg gggcgtaggg tcgccgacat gacacaaggg gttgacgcgt
      181 agtcgatcgc ggtaccgcag gaattccctg caggtcggag tactgtcctc cgagcggagt
      241 actgtcctcc gagcggagta ctgtcctccg agcggagtac tgtcctccga gcggagtact
      301 gtcctccgag cggagactct agcgagcgcc ggagtataaa tagaggcgct tcgtctacgg
      361 agcgacaatt caattcaaac aagcaaagtg aacacgtcgc taagcgaaag ctaagcaaat
      421 aaacaagcgc agctgaacaa gctaaacaat ctgcagtaaa gtgcaagtta aagtgaatca
      481 attaaaagta accagcaacc aagtaaatca actgcaacta ctgaaatctg ccaagaagta
      541 attattgaat acaagaagag aactctgaat agggaattgg gctcgagggt accagcgaag
      601 ttcctattcc gaagttccta ttctctagaa agtataggaa cttcgcagaa tggtagctgg
      661 attgtagctg ctattagcaa tatgaaacct cttagaagtt cctattccga agttcctatt
      721 cttcaaatag tataggaact tccctaggat cgatttaatt aagatccaga catgataaga
      781 tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt
      841 gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa acaagttaac
      901 aacaacaatt gcattcattt tatgtttcga gttcaggggg aggtgtggga ggttttttaa
      961 agcaagtaaa acctctacaa atgtggtatg gctgattatg atctctagtc aaggcactat
     1021 acatcaaata tttttccata attttcttgt atagcagtgc agctttttcc tttgtggtgt
     1081 aaatagcaaa gcaagcaaga gttctattac taaacacagc atgactcaaa aaacttagca
     1141 attctgaagg aaagtccttg gggtcttcta cctttctctt cttttttgga ggagtagaat
     1201 gttgagagtc agcagtagcc tcatcatcac tagatggcat ttcttctgag caaaacaggt
     1261 tttcctcatt aaaggcattc caccactgct cccattcatc agttccatag gttggaatct
     1321 aaaatacaca aacaattaga atcagtagtt taacacatta tacacttaaa aattttatat
     1381 ttaccttaga gctttaaatc tctgtaggta gtttgtccaa ttatgtcaca ccacagaagt
     1441 aaggttcctt cacaaagatc tcacttagtc aattttgtta gttagttaga cctgcggaag
     1501 agagataaat cggttgatta aaattagttc aagaataatt agtatattta aacatgccgg
     1561 ggatgggaca agtcgccatg ttggatcgac tttcgtacta gtaccggttt tcagatttat
     1621 ggggcttaca tacatacata attactttat acaacgaaaa ttactaaata gaatcgttcc
     1681 tgtaagcagg actgacacat ttgcttgcgt tgattgctat tatatcgacc taccatagtc
     1741 acataaatat gtacatatat aaatatgtat gtatgtacaa tcgcatgctt gacggcagtt
     1801 aagaaatgaa gtcaccattt cggtgagctg gcaacgatta tgctgtggtc ccgaaaggaa
     1861 tgactcatcg tcccagagaa tcgataccaa cggtggaaag ccaagtgcat gccacactgc
     1921 gtgcacagcc tgctgctaca cttgccacaa acagcaggcg gaaaagcgca gcagcggccc
     1981 gaatcgggga aaatagaaag cgctaattat agacgaagtg aacggcagct ctgctttccc
     2041 gcagtcgaaa ctccgtgcaa tgaccccgtt ctttcaccac gaaatgaatg atcgctggga
     2101 gaaagaaaag gcaaagaaaa acatagtggg gttttcttat ttctgacaac actgaatctg
     2161 gccgatttta ttcttgtgta cacaacttat cgccgagtta acttggcttt tctgctatac
     2221 gtgtctttgt ggataaaata ctttatggtt tctcgtacaa tcttcatcgt tttcgtatgc
     2281 ttttcagtgt tgcagtgaat tttttgttgc gattggaagc gtaaacactg atttcgacgg
     2341 ttaccccgcc caccaatgac gctcccatcg agcgttgaag tggcgcgacg cagatccaga
     2401 catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg
     2461 ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa
     2521 acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga
     2581 ggttttttaa agcaagtaaa acctctacaa atgtggtatg gctgattatg atctctagtc
     2641 aaggcactat acatcaaata tttttccata attttcttgt atagcagtgc agctttttcc
     2701 tttgtggtgt aaatagcaaa gcaagcaaga gttctattac taaacacagc atgactcaaa
     2761 aaacttagca attctgaagg aaagtccttg gggtcttcta cctttctctt cttttttgga
     2821 ggagtagaat gttgagagtc agcagtagcc tcatcatcac tagatggcat ttcttctgag
     2881 caaaacaggt tttcctcatt aaaggcattc caccactgct cccattcatc agttccatag
     2941 gttggaatct aaaatacaca aacaattaga atcagtagtt taacacatta tacacttaaa
     3001 aattttatat ttaccttaga gctttaaatc tctgtaggta gtttgtccaa ttatgtcaca
     3061 ccacagaagt aaggttcctt cacaaagatc gaggccggcc tcctacttat acagctcatc
     3121 catgccgtac agaaacaggt gatggcgtcc ctccgagcgc tcatactgct ccacaatcgt
     3181 atagtcctca ttgtgcgagg taatgtccag cttcgtatcc acatagtaat atcccggcag
     3241 ctgcactggc ttcttggcca tatagatcgt cttaaactcc accagatagc gtccgccatc
     3301 cttcagcttc agggcctggt ggatctcgcc cttcagcacg ccgtcgcggg ggtacaggcg
     3361 ctcggtcgag gcctcccatc ccatcgtctt cttctgcatc actgggccgt ccggcgggaa
     3421 attcgttccg cgcatcttca ctttatagat cagcgttccg tcctgcaggg aactgtcctg
     3481 cgtcacggtc acgagtccgc cgtcctcaaa attcatcacg cgctcccact tgaatccctc
     3541 tgggaacgac agcttcttat agtccggaat gtcggccggg tgcttcacat aggccttcga
     3601 gccatacata aactgcggcg acaggatgtc ccaggcaaac ggcagcggtc cgcccttcgt
     3661 cactttcagc ttggccgtct gcgtgccctc gtaggggcgt ccctcgccct cgccctcgat
     3721 ctcaaactca tgtccattca tggatccctc catgcgcact ttaaagcgca taaactcctt
     3781 aatcacggcc atgttgttat cctcgctgct ggcggttcca ctacttccgc ttccggtgct
     3841 gccggttccg tgtcccagaa acagatggtg gcggccctcg ctgcgctcgt actgctccac
     3901 gatggtgtaa tcctcgttgt ggctggtgat atccagcttg gtgtccacgt agtagtagcc
     3961 gggcagctgc acgggcttct tggccatgta gatggtcttg aactccacca ggtagtggcc
     4021 gccgtccttc agcttcaggg cctgatgaat ctcgcccttc agcacgccgt cgcgtggata
     4081 caggcgctcg gtgctggcct cccagcccat ggtcttcttc tgcatcacgg gtccatcggg
     4141 ggggaagttg gtgccgcgca tcttcacttt gtagatcagg gtgccatcct gcagactgct
     4201 atcctgggtc acggtcacga ggccgccatc ctcgaagttc atcacgcgct cccacttgaa
     4261 gccctcgggg aagctcagct tcttgtaatc ggggatatcg gcggggtgct tcacgtaggc
     4321 cttgctgccg tacataaact gggggctcag aatatcccag gcgaagggca ggggtccgcc
     4381 cttggtcact ttcagcttgg cggtctgggt tccctcatat gggcgtccct cgccctcgcc
     4441 ctcaatctcg aactcgtggc cgttcatgct gccctccatg cgcactttga agcgcatgaa
     4501 ctccttgatc acttcctcgc ccttggacac catagcggcc gcgaagttcc tatactttct
     4561 agagaatagg aacttcggaa taggaacttc ttgccttaac ccagaaatta tcactgttat
     4621 tctttgaaat ggtgcaaaga gaagttccta tactatttga agaataggaa cttcggaata
     4681 ggaacttcag gcctacgcgt attgggacat cggatccaag cttgtcgacg gtaccataac
     4741 ttcgtataat gtatgctata cgaagttatc gtacgacggt cgacggatct aattcaatta
     4801 gagactaatt caattagagc taattcaatt aggatccaag cttatcgatt tcgaaccctc
     4861 gaccgccgga gtataaatag aggcgcttcg tctacggagc gacaattcaa ttcaaacaag
     4921 caaagtgaac acgtcgctaa gcgaaagcta agcaaataaa caagcgcagc tgaacaagct
     4981 aaacaatcgg ctcgaagccg gtcgccacca tggcctcctc cgaggacgtc atcaaggagt
     5041 tcatgcgctt caaggtgcgc atggagggct ccgtgaacgg ccacgagttc gagatcgagg
     5101 gcgagggcga gggccgcccc tacgagggca cccagaccgc caagctgaag gtgaccaagg
     5161 gcggccccct gcccttcgcc tgggacatcc tgtcccccca gttccagtac ggctccaagg
     5221 tgtacgtgaa gcaccccgcc gacatccccg actacaagaa gctgtccttc cccgagggct
     5281 tcaagtggga gcgcgtgatg aacttcgagg acggcggcgt ggtgaccgtg acccaggact
     5341 cctccctcca ggacggctcc ttcatctaca aggtgaagtt catcggcgtg aacttcccct
     5401 ccgacggccc cgtaatgcag aagaagacta tgggctggga ggcgtccacc gagcgcctgt
     5461 acccccgcga cggcgtgctg aagggcgaga tccacaaggc cctgaagctg aaggacggcg
     5521 gccactacct ggtggagttc aagtccatct acatggccaa gaagcccgtg cagctgcccg
     5581 gctactacta cgtggactcc aagctggaca tcacctccca caacgaggac tacaccatcg
     5641 tggagcagta cgagcgcgcc gagggccgcc accacctgtt cctgtagggg ccgcgactct
     5701 agatcataat cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac
     5761 acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg
     5821 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     5881 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttag tcgacgcaac
     5941 cggtataact tcgtataatg tatgctatac gaagttatca accccttgtg tcatgtcggc
     6001 gaccctacgc ccccaactga gagaactcaa aggttacccc agttggggca ctactcccga
     6061 aaaccgcttc tgacctgggt atagaagagc gagctcacta gtaaagatct ccatgcataa
     6121 ggcgcgccta ggccttctgc agctcgaggc tcttccgtca tcggaaagaa catgtgagca
     6181 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     6241 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     6301 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     6361 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     6421 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     6481 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     6541 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     6601 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     6661 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     6721 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     6781 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     6841 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     6901 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     6961 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     7021 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     7081 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     7141 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     7201 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     7261 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     7321 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     7381 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     7441 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     7501 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     7561 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     7621 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     7681 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     7741 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     7801 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     7861 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     7921 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     7981 gacgtc
//
