LOCUS       sequence-468183-        5101 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  dsDNA donor vector with 3xP3-DsRed and FlpTag-GFP for precise 
            genomic targeting and integration of the FlpTag-GFP cassette with 
            codon phase 1..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5101)
  AUTHORS   Fendl S, Vieira RM, Borst A
  TITLE     Conditional protein tagging methods reveal highly specific 
            subcellular distribution of ion channels in motion-sensing neurons.
  JOURNAL   Elife. 2020 Oct 20;9:e62953. doi: 10.7554/eLife.62953.
  PUBMED    33079061
REFERENCE   2  (bases 1 to 5101)
  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: "2020-10-20"; volume: "9:e62953. doi"; pages: " 
            10.7554/eLife.62953"
COMMENT     Sequence Label: pHD-FlpTag-DsRed-HDR-ph1
FEATURES             Location/Qualifiers
     source          1..5101
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(41..74)
                     /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)"
     protein_bind    159..192
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     regulatory      418..427
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             424..1101
                     /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(554..572)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     complement(604..624)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     822..841
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1016..1039
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     polyA_signal    1222..1343
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1259..1278)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1313..1332
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    1350..1383
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    1403..1502
                     /label=phage phi-C31 attP
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attachment site of phage phi-C31"
     primer_bind     complement(1542..1559)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1542..1558)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1657..1674)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1828..2416)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1908..1927)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2538..3329,3330..3398))
                     /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     3161..3180
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3399..3490)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3532..3551
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3557..3576
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     protein_bind    3832..3931
                     /label=phage phi-C31 attP
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attachment site of phage phi-C31"
     protein_bind    4019..4052
                     /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)"
ORIGIN
        1 ccagaaatta tcactgttat tctttagaat ggtgcaaaga gaagttccta tactttctag
       61 agaataggaa cttcccgtct tcccgcggac atatgcacac ctgcgatcgt agtgccccaa
      121 ctggggtaac ctttgagttc tctcagttgg gggcgtagat aacttcgtat aatgtatgct
      181 atacgaagtt atcgtacggg atctaattca attagagact aattcaatta gagctaattc
      241 aattaggatc caagcttatc gatttcgaac cctcgaccgc cggagtataa atagaggcgc
      301 ttcgtctacg gagcgacaat tcaattcaaa caagcaaagt gaacacgtcg ctaagcgaaa
      361 gctaagcaaa taaacaagcg cagctgaaca agctaaacaa tcggctcgaa gccggtcgcc
      421 accatggcct cctccgagga cgtcatcaag gagttcatgc gcttcaaggt gcgcatggag
      481 ggctccgtga acggccacga gttcgagatc gagggcgagg gcgagggccg cccctacgag
      541 ggcacccaga ccgccaagct gaaggtgacc aagggcggcc ccctgccctt cgcctgggac
      601 atcctgtccc cccagttcca gtacggctcc aaggtgtacg tgaagcaccc cgccgacatc
      661 cccgactaca agaagctgtc cttccccgag ggcttcaagt gggagcgcgt gatgaacttc
      721 gaggacggcg gcgtggtgac cgtgacccag gactcctccc tccaggacgg ctccttcatc
      781 tacaaggtga agttcatcgg cgtgaacttc ccctccgacg gccccgtaat gcagaagaag
      841 actatgggct gggaggcgtc caccgagcgc ctgtaccccc gcgacggcgt gctgaagggc
      901 gagatccaca aggccctgaa gctgaaggac ggcggccact acctggtgga gttcaagtcc
      961 atctacatgg ccaagaagcc cgtgcagctg cccggctact actacgtgga ctccaagctg
     1021 gacatcacct cccacaacga ggactacacc atcgtggagc agtacgagcg cgccgagggc
     1081 cgccaccacc tgttcctgta ggggccgcga ctctagatca taatcagcca taccacattt
     1141 gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa
     1201 atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc
     1261 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     1321 tccaaactca tcaatgtatc ttaaccggta taacttcgta taatgtatgc tatacgaagt
     1381 tatagaagag cactccgtct tcaacccctt gtgtcatgtc ggcgacccta cgcccccaac
     1441 tgagagaact caaaggttac cccagttggg gcactactcc cgaaaaccgc ttctgacctg
     1501 ggaagcttgt tccatcccac tcgagtacgt aggtacctca gactggccgt cgttttacaa
     1561 cacagaaaga gtttgtagaa acgcaaaaag gccatccgtc aggggccttc tgcttagttt
     1621 gatgcctggc agttccctac tctcgccttc cgcttcctcg ctcactgact cgctgcgctc
     1681 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     1741 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     1801 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     1861 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     1921 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     1981 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     2041 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     2101 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     2161 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     2221 tgctacagag ttcttgaagt ggtgggctaa ctacggctac actagaagaa cagtatttgg
     2281 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     2341 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     2401 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     2461 cgacgcgcgc gtaactcacg ttaagggatt ttggtcatga gcttgcgccg tcccgtcaag
     2521 tcagcgtaat gctctgctta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     2581 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     2641 ggcttaccat ctggccccag cgctgcgatg ataccgcgag aaccacgctc accggctccg
     2701 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     2761 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     2821 gttaatagtt tgcgcaacgt tgttgccatc gctacaggca tcgtggtgtc acgctcgtcg
     2881 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     2941 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     3001 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     3061 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     3121 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     3181 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     3241 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     3301 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     3361 aagggaataa gggcgacacg gaaatgttga atactcatat tcttcctttt tcaatattat
     3421 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     3481 aataaacaaa taggggtcag tgttacaacc aattaaccaa ttctgaacat tatcgcgagc
     3541 ccatttatac ctgaatatgg ctcataacac cccttgtttg cctggcggca gtagcgcggt
     3601 ggtcccacct gaccccatgc cgaactcaga agtgaaacgc cgtagcgccg atggtagtgt
     3661 ggggactccc catgcgagag tagggaactg ccaggcatca aataaaacga aaggctcagt
     3721 cgaaagactg ggcctttcgc ccgggctaat tatggggtgt cgcccttcgc tgaagcaggt
     3781 ggaattcttg catgctccta ggtatactgc agttgctatc tattacaaag tcccaggtca
     3841 gaagcggttt tcgggagtag tgccccaact ggggtaacct ttgagttctc tcagttgggg
     3901 gcgtagggtc gccgacatga cacaaggggt tgaagacttc tgaagttcct attctatcag
     3961 aagtatagga acttcagcgg ctggattgta gctgctatta gcaatatgaa acctcttaga
     4021 agttcctatt ctctagaaag tataggaact tcgaattctg gcgacttgtc gcatctttct
     4081 ctgagtgggt aaggtcattg tggggtctta agggctaaag ggtagtgatg atgaaggcat
     4141 ttgatgccat tgttcaataa cttacctccg gaacctccgc ttccaccgct acctcccttg
     4201 tacagctcat ccatgcccag ggtgatgccg gcggcggtca cgaactccag cagcaccatg
     4261 tgatcgcgct tctcgttggg gtccttggac agcacgctct gggtgctcag gtagtggtta
     4321 tcgggcagca gcactgggcc gtcgccgatg ggggtgttct gctggtagtg atcggccagc
     4381 tgcacggagc catcctccac attgtggcgg atcttgaagt tggccttgat gccgttcttc
     4441 tgcttatcgg cggtgatgta cacgttgtgg ctgttgaagt tgtactccag cttgtggccc
     4501 aggatgttgc catcctcctt gaaatcgatg cccttcagct cgatgcggtt caccagggta
     4561 tcgccctcga acttcacctc ggcgcgggtc ttgtaggtgc cgtcatcctt gaagctgatg
     4621 gtgcgctcct gcacgtagcc ctcgggcatg gcgctcttga agaaatcgtg ctgcttcatg
     4681 tgatcggggt agcggctgaa gcactgcacg ccgtaggtca gggtggtcac cagggtgggc
     4741 cagggcacgg gcagcttgcc ggtggtgcag atgaacttca gggtcagctt gccgttggtg
     4801 gcgtcgccct cgccctcgcc gcgcacgctg aacttgtggc cgttcacgtc gccatccagc
     4861 tccaccagga tgggcaccac gccggtgaac agctcctcgc ccttggacac ggatccggat
     4921 ccgccgctac ctccgcttcc accggaacct ccgccacctg cggaagagag ataaatcggt
     4981 tgattaaaat tagttcaaga ataattagta tatttaaaca tgccggggat gggacaagtc
     5041 gccatgttgg atcgactgaa gttcctatac ttctgataga ataggaactt cttgccttaa
     5101 c
//
