LOCUS       sequence-378064-        7196 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  lyn-tagRFPt expression under the UAS promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7196)
  AUTHORS   Moller K, Brambach M, Villani A, Gallo E, Gilmour D, Peri F
  TITLE     A role for the centrosome in regulating the rate of neuronal 
            efferocytosis by microglia in vivo.
  JOURNAL   Elife. 2022 Nov 18;11:e82094. doi: 10.7554/eLife.82094.
  PUBMED    36398880
REFERENCE   2  (bases 1 to 7196)
  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: "2022-11-18"; volume: "11:e82094. doi"; pages: " 
            10.7554/eLife.82094"
COMMENT     Sequence Label: pDEST_6xUAS:lyn-tagRFPt
FEATURES             Location/Qualifiers
     source          1..7196
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(12..33)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     misc_feature    join(382..393,394..556)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     primer_bind     929..945
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    965..985
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     primer_bind     996..1012
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1028..1122
                     /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    1047..1141
                     /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    1221..1245
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      1296..1305
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(1372..1396)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1398..2111
                     /codon_start=1
                     /product="monomeric derivative of red fluorescent protein 
                     from Entacmaea quadricolor (Merzlyak et al., 2007)"
                     /label=TagRFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTM
                     RIKVVEGGPLPFAFDILATSFMYGSRTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEANTEMLYPADGGLEGRTD
                     MALKLVGGGHLICNFKTTYRSKKPAKNLKMPGVYYVDHRLERIKEADKETYVEQHEVAV
                     ARYCDLPSKLGHKLN"
     protein_bind    complement(2113..2133)
                     /gene="mutant version of attB"
                     /label=attB3
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     primer_bind     complement(2141..2157)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    2173..2307
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2197..2216)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2251..2270
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     polyA_signal    2450..2674
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     2663..2680
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(join(2703..3416,3417..3419,3420..3422))
                     /codon_start=1
                     /product="enhanced CFP"
                     /label=ECFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2748..2769)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3097..3116
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3356..3377
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      3419..3428
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    3581..3614
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     misc_feature    join(4594..4781,4782..4793)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(4827..4843)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4880..4900)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4880..4898)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4919..4935)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4932..4954)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4943..4959
                     /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(4967..4973,4974..4991,4992..4997))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5012..5033
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5150..5167)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5321..5909)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5401..5420)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6080..6871,6872..6940))
                     /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     6703..6722
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6941..7045)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca
       61 aagggcgaaa aaccgtctat cagggcgatg gcccacgctg gctaagaact catcagcctc
      121 cccggtccat ctacccacgt accaatgcac caattggcca caatgacggc tactacatgg
      181 tgccattcct tcctctttat aggaatggag actacctcct gtccaacaat gctcttggat
      241 acgagtacgc ctacctgttg gacccaggtc attgcacaac accagaaatg ccctctgatc
      301 tgcaaaagac gtgaatatct gttcagacac ccatatccac tctgttccac acaggtcaga
      361 ggtttgtcca ggagttcttg acagaggtgt aaaaagtact caaaaatttt actcaagtga
      421 aagtacaagt acttagggaa aattttactc aattaaaagt aaaagtatct ggctagaatc
      481 ttacttgagt aaaagtaaaa aagtactcca ttaaaattgt acttgagtat taaggaagta
      541 aaagtaaaag caagaaagaa aactagagat tcttgtttaa gcttttaatc tcaaaaaaca
      601 ttaaatgaaa tgcatacaag gttttatcct gctttagaac tgtttgtatt taattatcaa
      661 actataagac agacaatcta atgccagtac acgctactca aagttgtaaa acctcagatt
      721 taacttcagt agaagctgat tctcaaaatt gttagtgtca agcctagctc ttttggggct
      781 gaaaagcaat cctgcagtgc tgaaaagcct ctcacaggca gccgatgcgg gaagaggtgt
      841 attagtcttg atagagaggc tgcaaatagc aggaaacgtg agcagagact ccctggtgtc
      901 tgaaacacag gccagatggg ccctcgagca ggaaacagct atgaccatga ttacgccaag
      961 ctatcaactt tgtatagaaa agttgcggcg gccgctctag aactagtgga tcccccgggc
     1021 tgcaggtcgg agtactgtcc tccgagcgga gtactgtcct ccgagcggag tactgtcctc
     1081 cgagcggagt actgtcctcc gagcggagta ctgtcctccg agcggagtac tgtcctccga
     1141 gcggagactc tagagggtat ataatggatc ccatcgcgtc tcagcctcac tttgagctcc
     1201 tccacgaatt cgccgccacc gcaagtttgt acaaaaaagc aggctggatg ggctgcatca
     1261 agagcaagcg caaggacaac ctgaacgacg acgaggccgc catgggctgc atcaagagca
     1321 agcgcaagga caacctgaac gacgacgagg caccggaaga tcccgccacc gacccagctt
     1381 tcttgtacaa agtggctatg gtgtctaagg gcgaagagct gattaaggag aacatgcaca
     1441 tgaagctgta catggagggc accgtgaaca accaccactt caagtgcaca tccgagggcg
     1501 aaggcaagcc ctacgagggc acccagacca tgagaatcaa ggtggtcgag ggcggccctc
     1561 tccccttcgc cttcgacatc ctggctacca gcttcatgta cggcagcaga accttcatca
     1621 accacaccca gggcatcccc gacttcttta agcagtcctt ccctgagggc ttcacatggg
     1681 agagagtcac cacatacgaa gacgggggcg tgctgaccgc tacccaggac accagcctcc
     1741 aggacggctg cctcatctac aacgtcaaga tcagaggggt gaacttccca tccaacggcc
     1801 ctgtgatgca gaagaaaaca ctcggctggg aggccaacac cgagatgctg taccccgctg
     1861 acggcggcct ggaaggcaga accgacatgg ccctgaagct cgtgggcggg ggccacctga
     1921 tctgcaactt caagaccaca tacagatcca agaaacccgc taagaacctc aagatgcccg
     1981 gcgtctacta tgtggaccac agactggaaa gaatcaagga ggccgacaaa gagacctacg
     2041 tcgagcagca cgaggtggct gtggccagat actgcgacct ccctagcaaa ctggggcaca
     2101 aacttaatta agcaacttta ttatacatag ttgataattc actggccgtc gttttacggt
     2161 accatcgatg atgatccaga catgataaga tacattgatg agtttggaca aaccacaact
     2221 agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta
     2281 accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag
     2341 gttcaggggg aggtgtggga ggttttttaa agcaagtaaa acctctacaa atgtggtatg
     2401 gctgattatg atcctctaga tcagatcctg gttctttccg cctcagaagc catagagccc
     2461 accgcatccc cagcatgcct gctattgtct tcccaatcct cccccttgct gtcctgcccc
     2521 accccacccc ccagaataga atgacaccta ctcagacaat gcgatgcaat ttcctcattt
     2581 tattaggaaa ggacagtggg agtggcacct tccagggtca aggaaggcac gggggagggg
     2641 caaacaacag atggctggca actagaaggc acagtcgagg ctgatcagcg ggtttggccg
     2701 ctttacttgt acagctcgtc catgccgaga gtgatcccgg cggcggtcac gaactccagc
     2761 aggaccatgt gatcgcgctt ctcgttgggg tctttgctca gggcggactg ggtgctcagg
     2821 tagtggttgt cgggcagcag cacggggccg tcgccgatgg gggtgttctg ctggtagtgg
     2881 tcggcgagct gcacgctgcc gtcctcgatg ttgtggcgga tcttgaagtt ggccttgatg
     2941 ccgttcttct gcttgtcggc ggtgatatag acgttgtggc tgatgtagtt gtactccagc
     3001 ttgtgcccca ggatgttgcc gtcctccttg aagtcgatgc ccttcagctc gatgcggttc
     3061 accagggtgt cgccctcgaa cttcacctcg gcgcgggtct tgtagttgcc gtcgtccttg
     3121 aagaagatgg tgcgctcctg gacgtagcct tcgggcatgg cggacttgaa gaagtcgtgc
     3181 tgcttcatgt ggtcggggta gcggctgaag cactgcacgc cccaggtcag ggtggtcacg
     3241 agggtgggcc agggcacggg cagcttgccg gtggtgcaga tgaacttcag ggtcagcttg
     3301 ccgtaggtgg catcgccctc gccctcgccg gacacgctga acttgtggcc gtttacgtcg
     3361 ccgtccagct cgaccaggat gggcaccacc ccggtgaaca gctcctcgcc cttgctcacc
     3421 atggtggcgg gatcccctag caaaaataca gatactgcag tttctgatag gaaggttgag
     3481 cagaatagca ggggaaggaa acaggaggaa gacacttacc tgttaattaa cacggggaat
     3541 gcgcgttcca atgcaccgtt cccggcaacc tagcatgcag ataacttcgt ataaagtatc
     3601 ctatacgaag ttatggcaac gcgtttaaac tacagctgtg ctgggaaggc tggctcgctc
     3661 tggccttcct caccactgcc gcagctttta tacgctggca agggcagcac agcggcgcgc
     3721 ggcggacaat agaaacacgc cgtcatcaga gttcagccgt gccgaagcgc agggtcatcg
     3781 catcgcccag aaggccgtaa gcattttact gtcaatggaa agccctgacg gcggcacttt
     3841 ccctctcagc ccgagctcta tgctataggc tgtataccta tggctaagag aggtctgtgt
     3901 tcaggtcaga ggtcacgtga tgataatccg agcagaggag tcgctcgctg atacactcca
     3961 gagcagagta gatttcatac tgaagttaat aatgtagcat gacatatatt agatggtgtg
     4021 tgtgtgtgtg tgtatataca tatatatata tatatggtag cgtatttaga tggaatttat
     4081 tgatgtatta tttattatat gtatatacag atgaagtcaa aatttttcgc tccctgtata
     4141 ttttcccctt aaatcctgtt taagagaaag aaacatattt ctaatcataa tagttttaat
     4201 aactcatttc taataactga tttcttctct ctttgtcatg atgatagcac caattgctcg
     4261 agggggggcc cgatctgcga agatacggcc acgggtgctc ttgatcctgt ggctgatttt
     4321 ggactgtgct gctcgcagct gctgatgaat cacatacttc ctccattttc ttccactgat
     4381 tgactgttat aatttcccta atttccaggt caaggtgctg tgcattgtgg taatagatgt
     4441 gacatgacgt cacttccaaa ggaccaatga acatgtctga ccaatttcat ataatgtgaa
     4501 aacgattttc ataggcagaa taaataacat ttaaattaaa ctgggcatca gcgcaattca
     4561 attggtttgg taatagcaag ggaaaataga atgaagtgat ctccaaaaaa taagtacttt
     4621 ttgactgtaa ataaaattgt aaggagtaaa aagtactttt ttttctaaaa aaatgtaatt
     4681 aagtaaaagt aaaagtattg atttttaatt gtactcaagt aaagtaaaaa tccccaaaaa
     4741 taatacttaa gtacagtaat caagtaaaat tactcaagta ctttacacct ctggttcttg
     4801 accccctacc ttcagcaagc ccagcagatc cactagttct agagcggccg ccaccgcggt
     4861 ggagctccag cttttgttcc ctttagtgag ggttaattgc gcgcttggcg taatcatggt
     4921 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     4981 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     5041 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     5101 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     5161 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     5221 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     5281 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     5341 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     5401 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     5461 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     5521 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     5581 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     5641 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     5701 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     5761 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     5821 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     5881 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     5941 acgctcagtg gaacgaaaac tcacgttaag gcattttggt catgagatta tcaaaaagga
     6001 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     6061 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     6121 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     6181 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     6241 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     6301 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     6361 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     6421 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     6481 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     6541 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     6601 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     6661 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     6721 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     6781 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     6841 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     6901 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     6961 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     7021 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct aaattgtaag
     7081 cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca
     7141 ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggt
//
