LOCUS       sequence-468182-        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 0..
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-ph0
FEATURES             Location/Qualifiers
     source          1..5101
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    56..177
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(93..112)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     147..166
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    184..217
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    237..336
                     /label=phage phi-C31 attP
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attachment site of phage phi-C31"
     primer_bind     complement(376..393)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(376..392)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(491..508)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(662..1250)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(742..761)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1372..2163,2164..2232))
                     /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     1995..2014
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2233..2324)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2366..2385
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2391..2410
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     protein_bind    2666..2765
                     /label=phage phi-C31 attP
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attachment site of phage phi-C31"
     protein_bind    2853..2886
                     /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    complement(3976..4009)
                     /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    4094..4127
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     regulatory      4353..4362
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4359..5036
                     /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(4489..4507)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     complement(4539..4559)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     4757..4776
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     4951..4974
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
ORIGIN
        1 cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt
       61 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
      121 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttaacc
      181 ggtataactt cgtataatgt atgctatacg aagttataga agagcactcc gtcttcaacc
      241 ccttgtgtca tgtcggcgac cctacgcccc caactgagag aactcaaagg ttaccccagt
      301 tggggcacta ctcccgaaaa ccgcttctga cctgggaagc ttgttccatc ccactcgagt
      361 acgtaggtac ctcagactgg ccgtcgtttt acaacacaga aagagtttgt agaaacgcaa
      421 aaaggccatc cgtcaggggc cttctgctta gtttgatgcc tggcagttcc ctactctcgc
      481 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
      541 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
      601 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
      661 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
      721 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
      781 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
      841 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
      901 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
      961 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     1021 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggg
     1081 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     1141 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     1201 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     1261 tgatcttttc tacggggtct gacgctcagt ggaacgacgc gcgcgtaact cacgttaagg
     1321 gattttggtc atgagcttgc gccgtcccgt caagtcagcg taatgctctg cttaccaatg
     1381 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     1441 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagcgctgc
     1501 gatgataccg cgagaaccac gctcaccggc tccggattta tcagcaataa accagccagc
     1561 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     1621 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     1681 catcgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     1741 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     1801 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     1861 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     1921 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     1981 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     2041 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     2101 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     2161 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     2221 ttgaatactc atattcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     2281 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg tcagtgttac
     2341 aaccaattaa ccaattctga acattatcgc gagcccattt atacctgaat atggctcata
     2401 acaccccttg tttgcctggc ggcagtagcg cggtggtccc acctgacccc atgccgaact
     2461 cagaagtgaa acgccgtagc gccgatggta gtgtggggac tccccatgcg agagtaggga
     2521 actgccaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgcccgggc
     2581 taattatggg gtgtcgccct tcgctgaagc aggtggaatt cttgcatgct cctaggtata
     2641 ctgcagttgc tatctattac aaagtcccag gtcagaagcg gttttcggga gtagtgcccc
     2701 aactggggta acctttgagt tctctcagtt gggggcgtag ggtcgccgac atgacacaag
     2761 gggttgaaga cttctgaagt tcctattcta tcagaagtat aggaacttca gcggctggat
     2821 tgtagctgct attagcaata tgaaacctct tagaagttcc tattctctag aaagtatagg
     2881 aacttcgaat tctggcgact tgtcgcatct ttctctgagt gggtaaggtc attgtggggt
     2941 cttaagggct aaagggtagt gatgatgaag gcatttgatg ccattgttca ataacttacc
     3001 tcgccggaac ctccgcttcc accgctacct cccttgtaca gctcatccat gcccagggtg
     3061 atgccggcgg cggtcacgaa ctccagcagc accatgtgat cgcgcttctc gttggggtcc
     3121 ttggacagca cgctctgggt gctcaggtag tggttatcgg gcagcagcac tgggccgtcg
     3181 ccgatggggg tgttctgctg gtagtgatcg gccagctgca cggagccatc ctccacattg
     3241 tggcggatct tgaagttggc cttgatgccg ttcttctgct tatcggcggt gatgtacacg
     3301 ttgtggctgt tgaagttgta ctccagcttg tggcccagga tgttgccatc ctccttgaaa
     3361 tcgatgccct tcagctcgat gcggttcacc agggtatcgc cctcgaactt cacctcggcg
     3421 cgggtcttgt aggtgccgtc atccttgaag ctgatggtgc gctcctgcac gtagccctcg
     3481 ggcatggcgc tcttgaagaa atcgtgctgc ttcatgtgat cggggtagcg gctgaagcac
     3541 tgcacgccgt aggtcagggt ggtcaccagg gtgggccagg gcacgggcag cttgccggtg
     3601 gtgcagatga acttcagggt cagcttgccg ttggtggcgt cgccctcgcc ctcgccgcgc
     3661 acgctgaact tgtggccgtt cacgtcgcca tccagctcca ccaggatggg caccacgccg
     3721 gtgaacagct cctcgccctt ggacacggat ccggatccgc cgctacctcc gcttccaccg
     3781 gaacctccca cctgcggaag agagataaat cggttgatta aaattagttc aagaataatt
     3841 agtatattta aacatgccgg ggatgggaca agtcgccatg ttggatcgac tgaagttcct
     3901 atacttctga tagaatagga acttcttgcc ttaacccaga aattatcact gttattcttt
     3961 agaatggtgc aaagagaagt tcctatactt tctagagaat aggaacttcc cgtcttcccg
     4021 cggacatatg cacacctgcg atcgtagtgc cccaactggg gtaacctttg agttctctca
     4081 gttgggggcg tagataactt cgtataatgt atgctatacg aagttatcgt acgggatcta
     4141 attcaattag agactaattc aattagagct aattcaatta ggatccaagc ttatcgattt
     4201 cgaaccctcg accgccggag tataaataga ggcgcttcgt ctacggagcg acaattcaat
     4261 tcaaacaagc aaagtgaaca cgtcgctaag cgaaagctaa gcaaataaac aagcgcagct
     4321 gaacaagcta aacaatcggc tcgaagccgg tcgccaccat ggcctcctcc gaggacgtca
     4381 tcaaggagtt catgcgcttc aaggtgcgca tggagggctc cgtgaacggc cacgagttcg
     4441 agatcgaggg cgagggcgag ggccgcccct acgagggcac ccagaccgcc aagctgaagg
     4501 tgaccaaggg cggccccctg cccttcgcct gggacatcct gtccccccag ttccagtacg
     4561 gctccaaggt gtacgtgaag caccccgccg acatccccga ctacaagaag ctgtccttcc
     4621 ccgagggctt caagtgggag cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga
     4681 cccaggactc ctccctccag gacggctcct tcatctacaa ggtgaagttc atcggcgtga
     4741 acttcccctc cgacggcccc gtaatgcaga agaagactat gggctgggag gcgtccaccg
     4801 agcgcctgta cccccgcgac ggcgtgctga agggcgagat ccacaaggcc ctgaagctga
     4861 aggacggcgg ccactacctg gtggagttca agtccatcta catggccaag aagcccgtgc
     4921 agctgcccgg ctactactac gtggactcca agctggacat cacctcccac aacgaggact
     4981 acaccatcgt ggagcagtac gagcgcgccg agggccgcca ccacctgttc ctgtaggggc
     5041 cgcgactcta gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa
     5101 a
//
