LOCUS       sequence-378124-        7628 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  zfRabaa tagged with mNeonGreen, expressed under the UAS promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7628)
  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 7628)
  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:mNeonGreen-Rab5a
FEATURES             Location/Qualifiers
     source          1..7628
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(336..1049)
                     /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
                     VLTATQDTSLQDGCLIYNVKIRGANFPSNGPVMQKKTLGWEANTEMLYPADGGLEGRSD
                     MALKLVGGGHLICNFKTTYRSKKPAKNLKMPGVYYVDHRLERIKEADKETYVEQHEVAV
                     ARYYDLPSKLGHKLN"
     misc_feature    join(2272..2459,2460..2471)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(2505..2521)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2558..2578)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2558..2576)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2597..2613)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2610..2632)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2621..2637
                     /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(2645..2651,2652..2669,2670..2675))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2690..2711
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2828..2845)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2999..3587)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3079..3098)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3758..4549,4550..4618))
                     /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     4381..4400
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4619..4723)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4886..4907)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     misc_feature    join(5256..5267,5268..5430)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     primer_bind     5803..5819
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5839..5859
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     primer_bind     5870..5886
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5902..5996
                     /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    5921..6015
                     /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    6095..6119
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             6122..6829
                     /codon_start=1
                     /product="bright monomeric yellow-green fluorescent protein
                     derived from LanYFP (Shaner et al., 2013)"
                     /label=mNeonGreen
                     /translation="MVSKGEEDNMASLPATHELHIFGSINGVDFDMVGQGTGNPNDGYE
                     ELNLKSTKGDLQFSPWILVPHIGYGFHQYLPYPDGMSPFQAAMVDGSGYQVHRTMQFED
                     GASLTVNYRYTYEGSHIKGEAQVKGTGFPADGPVMTNSLTAADWCRSKKTYPNDKTIIS
                     TFKWSYTTGNGKRYRSTARTTYTFAKPMAANYLKNQPMYVFRKTELKHSKTELNFKEWQ
                     KAFTDVMGMDELYK"
     regulatory      6377..6386
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(6831..6855)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(7509..7529)
                     /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(7537..7553)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    join(7569..7628,1..75)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
ORIGIN
        1 aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc
       61 tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag
      121 gtgtgggagg ttttttaaag caagtaaaac ctctacaaat gtggtatggc tgattatgat
      181 cctctagatc agatctcttg tttattgcag cttataatgg ttacaaataa agcaatagca
      241 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
      301 tcatcaatgt atcttatcat gtctggatct acgtatcaat taagtttgtg ccccagtttg
      361 ctagggaggt cgtagtatct ggccacagcc acctcgtgct gctcgacgta ggtctctttg
      421 tcggcctcct tgattctttc cagtctgtgg tccacatagt agacaccggg catcttgagg
      481 ttcttagcgg gtttcttgga tctgtatgtg gtcttgaagt tgcagatcag gtggcccccg
      541 cccacgagct tcagggccat gtcgcttctg ccttccaggc cgccgtcagc ggggtacagc
      601 atctcggtgt tggcctccca gccgagtgtt ttcttctgca tcacagggcc gttggatggg
      661 aagttcgccc ctctgatctt gacgttgtag atgaggcagc cgtcctggag gctggtgtcc
      721 tgggtagcgg tcagcacgcc cccgtcttcg tatgtggtga ctctctccca tgtgaagccc
      781 tcagggaagg actgcttaaa gaagtcgggg atgccctggg tgtggttgat gaaggttctg
      841 ctgccgtaca tgaagctggt agccaggatg tcgaaggcga aggggagagg gccgccctcg
      901 accaccttga ttctcatggt ctgggtgccc tcgtagggct tgccttcgcc ctcggatgtg
      961 cacttgaagt ggtggttgtt cacggtgccc tccatgtaca gcttcatgtg catgttctcc
     1021 ttaatcagct cttcgccctt agacaccatg gtcactgtct gctttgctgt tggtctgggc
     1081 tcctgggtca ctggcttact aatggagtct ttatgtatga ggactcttat caattgttct
     1141 tctataaagg tctgcagtgt ttctgttcgt cccctacatg gacacccaga gcctcctaaa
     1201 tacaggagcc ctgataactg cacaagtgct cagattccag cagggtggaa aatgagataa
     1261 agtgtgcaga tggggagggg gacgtgaatg agagatttga gggatgaaaa ggatggatga
     1321 acgcattgaa aatagcccct ttcacaaaat aataccagta aattgtcata taattaatga
     1381 ccagaccttt acctgtaaat gatgtgatgt tcatatacag aagaatgcca acaatttaca
     1441 gatgatttta caaattctta attttccttt tctaaatgga tttgttcaca cattatctct
     1501 tcaaagcaat tgaattttct ggaaaagatt ggctgtgtgt gtaaaaggtg ctactgagga
     1561 atttaatgtg acatggaatg aagcaaacag cagcctaaga caggatggca ggaaaaatgt
     1621 cacatgttta aaataagagt gaaaccaacg acctggactt atagagactg tgtgtcctta
     1681 ggcaaagctc ttgtacaaac agtgcttgtg actgtgaaat tcggggtttg cctggattgt
     1741 gttaaatgtg tgcttggatg tatcataaaa aagaatcggt tctctgtatt tgacattttc
     1801 tgtaatttat ctaaagttac aactggctat gcctgatttt tatcactatt gaatcaatca
     1861 ttttaaagaa tgttaagtct gctgatttgg agagctcatt tacatttatt ttgaatgtct
     1921 cttatttaac acaactgatt taagctttag atctgcgaag atacggccac gggtgctctt
     1981 gatcctgtgg ctgattttgg actgtgctgc tcgcagctgc tgatgaatca catacttcct
     2041 ccattttctt ccactgattg actgttataa tttccctaat ttccaggtca aggtgctgtg
     2101 cattgtggta atagatgtga catgacgtca cttccaaagg accaatgaac atgtctgacc
     2161 aatttcatat aatgtgaaaa cgattttcat aggcagaata aataacattt aaattaaact
     2221 gggcatcagc gcaattcaat tggtttggta atagcaaggg aaaatagaat gaagtgatct
     2281 ccaaaaaata agtacttttt gactgtaaat aaaattgtaa ggagtaaaaa gtactttttt
     2341 ttctaaaaaa atgtaattaa gtaaaagtaa aagtattgat ttttaattgt actcaagtaa
     2401 agtaaaaatc cccaaaaata atacttaagt acagtaatca agtaaaatta ctcaagtact
     2461 ttacacctct ggttcttgac cccctacctt cagcaagccc agcagatcca ctagttctag
     2521 agcggccgcc accgcggtgg agctccagct tttgttccct ttagtgaggg ttaattgcgc
     2581 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     2641 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     2701 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     2761 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     2821 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     2881 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     2941 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     3001 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     3061 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     3121 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     3181 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     3241 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     3301 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     3361 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     3421 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     3481 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     3541 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     3601 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggc attttggtca
     3661 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     3721 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     3781 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     3841 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     3901 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     3961 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     4021 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     4081 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     4141 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     4201 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     4261 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     4321 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     4381 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     4441 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     4501 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     4561 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     4621 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     4681 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     4741 tgccacctaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat
     4801 cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata
     4861 gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt
     4921 ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac gctggctaag
     4981 aactcatcag cctccccggt ccatctaccc acgtaccaat gcaccaattg gccacaatga
     5041 cggctactac atggtgccat tccttcctct ttataggaat ggagactacc tcctgtccaa
     5101 caatgctctt ggatacgagt acgcctacct gttggaccca ggtcattgca caacaccaga
     5161 aatgccctct gatctgcaaa agacgtgaat atctgttcag acacccatat ccactctgtt
     5221 ccacacaggt cagaggtttg tccaggagtt cttgacagag gtgtaaaaag tactcaaaaa
     5281 ttttactcaa gtgaaagtac aagtacttag ggaaaatttt actcaattaa aagtaaaagt
     5341 atctggctag aatcttactt gagtaaaagt aaaaaagtac tccattaaaa ttgtacttga
     5401 gtattaagga agtaaaagta aaagcaagaa agaaaactag agattcttgt ttaagctttt
     5461 aatctcaaaa aacattaaat gaaatgcata caaggtttta tcctgcttta gaactgtttg
     5521 tatttaatta tcaaactata agacagacaa tctaatgcca gtacacgcta ctcaaagttg
     5581 taaaacctca gatttaactt cagtagaagc tgattctcaa aattgttagt gtcaagccta
     5641 gctcttttgg ggctgaaaag caatcctgca gtgctgaaaa gcctctcaca ggcagccgat
     5701 gcgggaagag gtgtattagt cttgatagag aggctgcaaa tagcaggaaa cgtgagcaga
     5761 gactccctgg tgtctgaaac acaggccaga tgggccctcg agcaggaaac agctatgacc
     5821 atgattacgc caagctatca actttgtata gaaaagttgc ggcggccgct ctagaactag
     5881 tggatccccc gggctgcagg tcggagtact gtcctccgag cggagtactg tcctccgagc
     5941 ggagtactgt cctccgagcg gagtactgtc ctccgagcgg agtactgtcc tccgagcgga
     6001 gtactgtcct ccgagcggag actctagagg gtatataatg gatcccatcg cgtctcagcc
     6061 tcactttgag ctcctccacg aattcgccgc caccgcaagt ttgtacaaaa aagcaggctg
     6121 gatggtgagc aagggcgagg aggataacat ggcctctctc ccagcgacac atgagttaca
     6181 catctttggc tccatcaacg gtgtggactt tgacatggtg ggtcagggca ccggcaatcc
     6241 aaatgatggt tatgaggagt taaacctgaa gtccaccaag ggtgacctcc agttctcccc
     6301 ctggattctg gtccctcata tcgggtatgg cttccatcag tacctgccct accctgacgg
     6361 gatgtcgcct ttccaggccg ccatggtaga tggctccgga taccaagtcc atcgcacaat
     6421 gcagtttgaa gatggtgcct cccttactgt taactaccgc tacacctacg agggaagcca
     6481 catcaaagga gaggcccagg tgaaggggac tggtttccct gctgacggtc ctgtgatgac
     6541 caactcgctg accgctgcgg actggtgcag gtcgaagaag acttacccca acgacaaaac
     6601 catcatcagt acctttaagt ggagttacac cactggaaat ggcaagcgct accggagcac
     6661 tgcgcggacc acctacacct ttgccaagcc aatggcggct aactatctga agaaccagcc
     6721 gatgtacgtg ttccgtaaga cggagctcaa gcactccaag accgagctca acttcaagga
     6781 gtggcaaaag gcctttaccg atgtgatggg catggacgag ctgtacaagg acccagcttt
     6841 cttgtacaaa gtggctatgg ccaatagggg aggagcaaca cgccccaacg ggtccaacgc
     6901 gggcaataag atctgccagt ttaaactggt cttgctagga gagtcagcgg tggggaagtc
     6961 cagcctcgtt ctgcgcttcg tcaaagggca gtttcacgag tttcaggaaa gcacaatagg
     7021 agcggccttt cttacacaga cactgtgctt ggatgacacg acggtaaagt ttgagatctg
     7081 ggacacagct ggacaggagc gctaccacag tctggccccc atgtactaca gaggtgccca
     7141 ggccgccatt gtagtttatg acatcaccaa tgaggagtca tttgcaagag caaagaactg
     7201 ggttaaagag cttcagaggc aggctagccc aaacattgtc atcgctctgt ctgggaacaa
     7261 ggctgacctt gccaacaaga gagctgtgga cttccaggat gctcagtctt atgcagatga
     7321 caacagtttg ctgttcatgg agacgtcagc aaaaacctcc atgaacgtga acgagatctt
     7381 catggccatt gcgaaaaagt tgcctaagag tgagccccaa gctgccggag ccaacagcgg
     7441 gcggagtagg ggtgtggacc tcacagagac agcccaaccc actaaagccc cctgctgcag
     7501 caactaagca actttattat acatagttga taattcactg gccgtcgttt tacggtacca
     7561 tcgatgatga tccagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa
     7621 tgcagtga
//
