LOCUS       sequence-378068-        7537 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  zfCETN4 tagged with miRFP670, expressed under the UAS promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7537)
  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 7537)
  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:miRFP670-cetn4
FEATURES             Location/Qualifiers
     source          1..7537
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    33..167
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(57..76)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     111..130
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     protein_bind    complement(938..971)
                     /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)."
     regulatory      1124..1133
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1130..1828
                     /codon_start=1
                     /product="monomeric far-red fluorescent protein (Shcherbo 
                     et al., 2009)"
                     /label=mKate2
                     /note="mammalian codon-optimized"
                     /translation="MVSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMRIK
                     AVEGGPLPFAFDILATSFMYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLT
                     ATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEASTETLYPADGGLEGRADMAL
                     KLVGGGHLICNLKTTYRSKKPAKNLKMPGVYYVDRRLERIKEADKETYVEQHEVAVARY
                     CDLPSKLGHR"
     misc_feature    join(2152..2339,2340..2351)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(2385..2401)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2438..2458)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2438..2456)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2477..2493)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2490..2512)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2501..2517
                     /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(2525..2531,2532..2549,2550..2555))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2570..2591
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2708..2725)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2879..3467)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2959..2978)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3638..4429,4430..4498))
                     /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     4261..4280
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4499..4603)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4766..4787)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     misc_feature    join(5136..5147,5148..5310)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     primer_bind     5683..5699
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5719..5739
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     primer_bind     5750..5766
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5782..5876
                     /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    5801..5895
                     /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    5975..5999
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      6005..6014
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             6011..6955
                     /codon_start=1
                     /product="bright monomeric near-infrared fluorescent 
                     protein engineered from a bacterial phytochrome, with an 
                     emission maximum of 670 nm (Shcherbakova et al., 2016)"
                     /label=miRFP670
                     /label=ORF frame 1
                     /translation="MVAGHASGSPAFGTASHSNCEHEEIHLAGSIQPHGALLVVSEHDH
                     RVIQASANAAEFLNLGSVLGVPLAEIDGDLLIKILPHLDPTAEGMPVAVRCRIGNPSTE
                     YCGLMHRPPEGGLIIELERAGPSIDLSGTLAPALERIRTAGSLRALCDDTVLLFQQCTG
                     YDRVMVYRFDEQGHGLVFSECHVPGLESYFGNRYPSSTVPQMARQLYVRQRVRVLVDVT
                     YQPVPLEPRLSPLTGRDLDMSGCFLRSMSPCHLQFLKDMGVRATLAVSLVVGGKLWGLV
                     VCHHYLPRFIRFELRAICKRLAERIATRITALES"
     protein_bind    complement(6957..6981)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
ORIGIN
        1 actgrycgtc gtwttacggt accatcgatg wwgatccaga catgataaga tacattgatg
       61 agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg
      121 atgctattgc tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt
      181 gcattcattt tatgtttcag gttcaggggg aggtgtggga ggttttttaa agcaagtaaa
      241 acctctacaa atgtggtatg gctgattatg atcctctaga tcagatctcc tcgagcaatt
      301 ggtgctatca tcatgacaaa gagagaagaa atcagttatt agaaatgagt tattaaaact
      361 attatgatta gaaatatgtt tctttctctt aaacaggatt taaggggaaa atatacaggg
      421 agcgaaaaat tttgacttca tctgtatata catataataa ataatacatc aataaattcc
      481 atctaaatac gctaccatat atatatatat atgtatatac acacacacac acacaccatc
      541 taatatatgt catgctacat tattaacttc agtatgaaat ctactctgct ctggagtgta
      601 tcagcgagcg actcctctgc tcggattatc atcacgtgac ctctgacctg aacacagacc
      661 tctcttagcc ataggtatac agcctatagc atagagctcg ggctgagagg gaaagtgccg
      721 ccgtcagggc tttccattga cagtaaaatg cttacggcct tctgggcgat gcgatgaccc
      781 tgcgcttcgg cacggctgaa ctctgatgac ggcgtgtttc tattgtccgc cgcgcgccgc
      841 tgtgctgccc ttgccagcgt ataaaagctg cggcagtggt gaggaaggcc agagcgagcc
      901 agccttccca gcacagctgt agtttaaacg cgttgccata acttcgtata ggatacttta
      961 tacgaagtta tctgcatgct aggttgccgg gaacggtgca ttggaacgcg cattccccgt
     1021 gttaattaac aggtaagtgt cttcctcctg tttccttccc ctgctattct gctcaacctt
     1081 cctatcagaa actgcagtat ctgtattttt gctaggggat cccgccacca tggtgagcga
     1141 gctgattaag gagaacatgc acatgaagct gtacatggag ggcaccgtga acaaccacca
     1201 cttcaagtgc acatccgagg gcgaaggcaa gccctacgag ggcacccaga ccatgagaat
     1261 caaggcggtc gagggcggcc ctctcccctt cgccttcgac atcctggcta ccagcttcat
     1321 gtacggcagc aaaaccttca tcaaccacac ccagggcatc cccgacttct ttaagcagtc
     1381 cttccccgag ggcttcacat gggagagagt caccacatac gaagacgggg gcgtgctgac
     1441 cgctacccag gacaccagcc tccaggacgg ctgcctcatc tacaacgtca agatcagagg
     1501 ggtgaacttc ccatccaacg gccctgtgat gcagaagaaa acactcggct gggaggcctc
     1561 caccgagacc ctgtaccccg ctgacggcgg cctggaaggc agagccgaca tggccctgaa
     1621 gctcgtgggc gggggccacc tgatctgcaa cttgaagacc acatacagat ccaagaaacc
     1681 cgctaagaac ctcaagatgc ccggcgtcta ctatgtggac agaagactgg aaagaatcaa
     1741 ggaggccgac aaagagacct acgtcgagca gcacgaggtg gctgtggcca gatactgcga
     1801 cctccctagc aaactggggc acagatgaag atctgcgaag atacggccac gggtgctctt
     1861 gatcctgtgg ctgattttgg actgtgctgc tcgcagctgc tgatgaatca catacttcct
     1921 ccattttctt ccactgattg actgttataa tttccctaat ttccaggtca aggtgctgtg
     1981 cattgtggta atagatgtga catgacgtca cttccaaagg accaatgaac atgtctgacc
     2041 aatttcatat aatgtgaaaa cgattttcat aggcagaata aataacattt aaattaaact
     2101 gggcatcagc gcaattcaat tggtttggta atagcaaggg aaaatagaat gaagtgatct
     2161 ccaaaaaata agtacttttt gactgtaaat aaaattgtaa ggagtaaaaa gtactttttt
     2221 ttctaaaaaa atgtaattaa gtaaaagtaa aagtattgat ttttaattgt actcaagtaa
     2281 agtaaaaatc cccaaaaata atacttaagt acagtaatca agtaaaatta ctcaagtact
     2341 ttacacctct ggttcttgac cccctacctt cagcaagccc agcagatcca ctagttctag
     2401 agcggccgcc accgcggtgg agctccagct tttgttccct ttagtgaggg ttaattgcgc
     2461 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     2521 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     2581 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     2641 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     2701 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     2761 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     2821 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     2881 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     2941 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     3001 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     3061 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     3121 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     3181 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     3241 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     3301 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     3361 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     3421 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     3481 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggc attttggtca
     3541 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     3601 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     3661 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     3721 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     3781 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     3841 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     3901 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     3961 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     4021 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     4081 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     4141 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     4201 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     4261 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     4321 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     4381 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     4441 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     4501 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     4561 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     4621 tgccacctaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat
     4681 cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata
     4741 gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt
     4801 ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac gctggctaag
     4861 aactcatcag cctccccggt ccatctaccc acgtaccaat gcaccaattg gccacaatga
     4921 cggctactac atggtgccat tccttcctct ttataggaat ggagactacc tcctgtccaa
     4981 caatgctctt ggatacgagt acgcctacct gttggaccca ggtcattgca caacaccaga
     5041 aatgccctct gatctgcaaa agacgtgaat atctgttcag acacccatat ccactctgtt
     5101 ccacacaggt cagaggtttg tccaggagtt cttgacagag gtgtaaaaag tactcaaaaa
     5161 ttttactcaa gtgaaagtac aagtacttag ggaaaatttt actcaattaa aagtaaaagt
     5221 atctggctag aatcttactt gagtaaaagt aaaaaagtac tccattaaaa ttgtacttga
     5281 gtattaagga agtaaaagta aaagcaagaa agaaaactag agattcttgt ttaagctttt
     5341 aatctcaaaa aacattaaat gaaatgcata caaggtttta tcctgcttta gaactgtttg
     5401 tatttaatta tcaaactata agacagacaa tctaatgcca gtacacgcta ctcaaagttg
     5461 taaaacctca gatttaactt cagtagaagc tgattctcaa aattgttagt gtcaagccta
     5521 gctcttttgg ggctgaaaag caatcctgca gtgctgaaaa gcctctcaca ggcagccgat
     5581 gcgggaagag gtgtattagt cttgatagag aggctgcaaa tagcaggaaa cgtgagcaga
     5641 gactccctgg tgtctgaaac acaggccaga tgggccctcg agcaggaaac agctatgacc
     5701 atgattacgc caagctatca actttgtata gaaaagttgc ggcggccgct ctagaactag
     5761 tggatccccc gggctgcagg tcggagtact gtcctccgag cggagtactg tcctccgagc
     5821 ggagtactgt cctccgagcg gagtactgtc ctccgagcgg agtactgtcc tccgagcgga
     5881 gtactgtcct ccgagcggag actctagagg gtatataatg gatcccatcg cgtctcagcc
     5941 tcactttgag ctcctccacg aattcgccgc caccgcaagt ttgtacaaaa aagcaggctt
     6001 agccgccacc atggtagcag gtcatgcctc tggcagcccc gcattcggga ccgcctctca
     6061 ttcgaattgc gaacatgaag agatccacct cgccggctcg atccagccgc atggcgcgct
     6121 tctggtcgtc agcgaacatg atcatcgcgt catccaggcc agcgccaacg ccgcggaatt
     6181 tctgaatctc ggaagcgtac tcggcgttcc gctcgccgag atcgacggcg atctgttgat
     6241 caagatcctg ccgcatctcg atcccaccgc cgaaggcatg ccggtcgcgg tgcgctgccg
     6301 gatcggcaat ccctctacgg agtactgcgg tctgatgcat cggcctccgg aaggcgggct
     6361 gatcatcgaa ctcgaacgtg ccggcccgtc gatcgatctg tcaggcacgc tggcgccggc
     6421 gctggagcgg atccgcacgg cgggttcact gcgcgcgctg tgcgatgaca ccgtgctgct
     6481 gtttcagcag tgcaccggct acgaccgggt gatggtgtat cgtttcgatg agcaaggcca
     6541 cggcctggta ttctccgagt gccatgtgcc tgggctcgaa tcctatttcg gcaaccgcta
     6601 tccgtcgtcg actgtcccgc agatggcgcg gcagctgtac gtgcggcagc gcgtccgcgt
     6661 gctggtcgac gtcacctatc agccggtgcc gctggagccg cggctgtcgc cgctgaccgg
     6721 gcgcgatctc gacatgtcgg gctgcttcct gcgctcgatg tcgccgtgcc atctgcagtt
     6781 cctgaaggac atgggcgtgc gcgccaccct ggcggtgtcg ctggtggtcg gcggcaagct
     6841 gtggggcctg gttgtctgtc accattatct gccgcgcttc atccgtttcg agctgcgggc
     6901 gatctgcaaa cggctcgccg aaaggatcgc gacgcggatc accgcgcttg agagcaaccc
     6961 agctttcttg tacaaagtgg ggggtgctcg agcgatggcg tccggcttca ggaaaagcag
     7021 tgcttcagca aaccagagga agaaagcagg acccaaacca gagctgacag aagagcagaa
     7081 gcaggagatc aaagaggcgt tcgatctgtt cgacacagac ggctctggaa ccatcgatgt
     7141 caaggaactg aaggttgcga tgcgagctct ggggtttgag ccgaagaaag aagagataaa
     7201 gaagatgatc gctgatattg ataaagaagg atcgggcgta atcggcttca gcgatttcct
     7261 ttctatgatg acacagaaaa tgagtgagaa agactcaaaa gaagagatcc tgaaggcttt
     7321 ccggctgttt gacgatgact gcactgggaa aatctccttc aagaacctga agcgagtggc
     7381 taaagagctc ggagagaacc tgacggatga ggagctgcag gaaatgatcg acgaggccga
     7441 cagagacggt gacggggaga ttaatgagca ggagtttctg cggattatga agaaaaccaa
     7501 tctgtactaa wctnkwayta taswkagtyg wwwwwys
//
