LOCUS       sequence-468184-        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 2..
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-ph2
FEATURES             Location/Qualifiers
     source          1..5101
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(52..85)
                     /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    170..203
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     regulatory      429..438
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             435..1112
                     /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(565..583)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     complement(615..635)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     833..852
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1027..1050
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     polyA_signal    1233..1354
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1270..1289)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1324..1343
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    1361..1394
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    1414..1513
                     /label=phage phi-C31 attP
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attachment site of phage phi-C31"
     primer_bind     complement(1553..1570)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1553..1569)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1668..1685)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1839..2427)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1919..1938)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2549..3340,3341..3409))
                     /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     3172..3191
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3410..3501)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3543..3562
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3568..3587
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     protein_bind    3843..3942
                     /label=phage phi-C31 attP
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attachment site of phage phi-C31"
     protein_bind    4030..4063
                     /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 cttgccttaa cccagaaatt atcactgtta ttctttagaa tggtgcaaag agaagttcct
       61 atactttcta gagaatagga acttcccgtc ttcccgcgga catatgcaca cctgcgatcg
      121 tagtgcccca actggggtaa cctttgagtt ctctcagttg ggggcgtaga taacttcgta
      181 taatgtatgc tatacgaagt tatcgtacgg gatctaattc aattagagac taattcaatt
      241 agagctaatt caattaggat ccaagcttat cgatttcgaa ccctcgaccg ccggagtata
      301 aatagaggcg cttcgtctac ggagcgacaa ttcaattcaa acaagcaaag tgaacacgtc
      361 gctaagcgaa agctaagcaa ataaacaagc gcagctgaac aagctaaaca atcggctcga
      421 agccggtcgc caccatggcc tcctccgagg acgtcatcaa ggagttcatg cgcttcaagg
      481 tgcgcatgga gggctccgtg aacggccacg agttcgagat cgagggcgag ggcgagggcc
      541 gcccctacga gggcacccag accgccaagc tgaaggtgac caagggcggc cccctgccct
      601 tcgcctggga catcctgtcc ccccagttcc agtacggctc caaggtgtac gtgaagcacc
      661 ccgccgacat ccccgactac aagaagctgt ccttccccga gggcttcaag tgggagcgcg
      721 tgatgaactt cgaggacggc ggcgtggtga ccgtgaccca ggactcctcc ctccaggacg
      781 gctccttcat ctacaaggtg aagttcatcg gcgtgaactt cccctccgac ggccccgtaa
      841 tgcagaagaa gactatgggc tgggaggcgt ccaccgagcg cctgtacccc cgcgacggcg
      901 tgctgaaggg cgagatccac aaggccctga agctgaagga cggcggccac tacctggtgg
      961 agttcaagtc catctacatg gccaagaagc ccgtgcagct gcccggctac tactacgtgg
     1021 actccaagct ggacatcacc tcccacaacg aggactacac catcgtggag cagtacgagc
     1081 gcgccgaggg ccgccaccac ctgttcctgt aggggccgcg actctagatc ataatcagcc
     1141 ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc
     1201 tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt
     1261 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     1321 gttgtggttt gtccaaactc atcaatgtat cttaaccggt ataacttcgt ataatgtatg
     1381 ctatacgaag ttatagaaga gcactccgtc ttcaacccct tgtgtcatgt cggcgaccct
     1441 acgcccccaa ctgagagaac tcaaaggtta ccccagttgg ggcactactc ccgaaaaccg
     1501 cttctgacct gggaagcttg ttccatccca ctcgagtacg taggtacctc agactggccg
     1561 tcgttttaca acacagaaag agtttgtaga aacgcaaaaa ggccatccgt caggggcctt
     1621 ctgcttagtt tgatgcctgg cagttcccta ctctcgcctt ccgcttcctc gctcactgac
     1681 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     1741 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     1801 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     1861 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     1921 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     1981 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     2041 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     2101 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     2161 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     2221 tatgtaggcg gtgctacaga gttcttgaag tggtgggcta actacggcta cactagaaga
     2281 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     2341 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     2401 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     2461 gctcagtgga acgacgcgcg cgtaactcac gttaagggat tttggtcatg agcttgcgcc
     2521 gtcccgtcaa gtcagcgtaa tgctctgctt accaatgctt aatcagtgag gcacctatct
     2581 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     2641 cgatacggga gggcttacca tctggcccca gcgctgcgat gataccgcga gaaccacgct
     2701 caccggctcc ggatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     2761 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     2821 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat cgctacaggc atcgtggtgt
     2881 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     2941 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     3001 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     3061 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     3121 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     3181 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     3241 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     3301 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     3361 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ttcttccttt
     3421 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     3481 gtatttagaa aaataaacaa ataggggtca gtgttacaac caattaacca attctgaaca
     3541 ttatcgcgag cccatttata cctgaatatg gctcataaca ccccttgttt gcctggcggc
     3601 agtagcgcgg tggtcccacc tgaccccatg ccgaactcag aagtgaaacg ccgtagcgcc
     3661 gatggtagtg tggggactcc ccatgcgaga gtagggaact gccaggcatc aaataaaacg
     3721 aaaggctcag tcgaaagact gggcctttcg cccgggctaa ttatggggtg tcgcccttcg
     3781 ctgaagcagg tggaattctt gcatgctcct aggtatactg cagttgctat ctattacaaa
     3841 gtcccaggtc agaagcggtt ttcgggagta gtgccccaac tggggtaacc tttgagttct
     3901 ctcagttggg ggcgtagggt cgccgacatg acacaagggg ttgaagactt ctgaagttcc
     3961 tattctatca gaagtatagg aacttcagcg gctggattgt agctgctatt agcaatatga
     4021 aacctcttag aagttcctat tctctagaaa gtataggaac ttcgaattct ggcgacttgt
     4081 cgcatctttc tctgagtggg taaggtcatt gtggggtctt aagggctaaa gggtagtgat
     4141 gatgaaggca tttgatgcca ttgttcaata acttacctgc cggaacctcc gcttccaccg
     4201 ctacctccct tgtacagctc atccatgccc agggtgatgc cggcggcggt cacgaactcc
     4261 agcagcacca tgtgatcgcg cttctcgttg gggtccttgg acagcacgct ctgggtgctc
     4321 aggtagtggt tatcgggcag cagcactggg ccgtcgccga tgggggtgtt ctgctggtag
     4381 tgatcggcca gctgcacgga gccatcctcc acattgtggc ggatcttgaa gttggccttg
     4441 atgccgttct tctgcttatc ggcggtgatg tacacgttgt ggctgttgaa gttgtactcc
     4501 agcttgtggc ccaggatgtt gccatcctcc ttgaaatcga tgcccttcag ctcgatgcgg
     4561 ttcaccaggg tatcgccctc gaacttcacc tcggcgcggg tcttgtaggt gccgtcatcc
     4621 ttgaagctga tggtgcgctc ctgcacgtag ccctcgggca tggcgctctt gaagaaatcg
     4681 tgctgcttca tgtgatcggg gtagcggctg aagcactgca cgccgtaggt cagggtggtc
     4741 accagggtgg gccagggcac gggcagcttg ccggtggtgc agatgaactt cagggtcagc
     4801 ttgccgttgg tggcgtcgcc ctcgccctcg ccgcgcacgc tgaacttgtg gccgttcacg
     4861 tcgccatcca gctccaccag gatgggcacc acgccggtga acagctcctc gcccttggac
     4921 acggatccgg atccgccgct acctccgctt ccaccggaac ctcccgacct gcggaagaga
     4981 gataaatcgg ttgattaaaa ttagttcaag aataattagt atatttaaac atgccgggga
     5041 tgggacaagt cgccatgttg gatcgactga agttcctata cttctgatag aataggaact
     5101 t
//
