LOCUS       sequence-208091-        5865 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Codon-optimized monomeric near-infrared fluorescent protein miRFP in
            zebrafish expression plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5865)
  AUTHORS   Piatkevich KD, Jung EE, Straub C, Linghu C, Park D, Suk HJ, Hochbaum
            DR, Goodwin D, Pnevmatikakis E, Pak N, Kawashima T, Yang CT, Rhoades
            JL, Shemesh O, Asano S, Yoon YG, Freifeld L, Saulnier JL, Riegler C,
            Engert F, Hughes T, Drobizhev M, Szabo B, Ahrens MB, Flavell SW, 
            Sabatini BL, Boyden ES
  TITLE     A robotic multidimensional directed evolution approach applied to 
            fluorescent voltage reporters.
  JOURNAL   Nat Chem Biol. 2018 Feb 26. pii: 10.1038/s41589-018-0004-9. doi: 
            10.1038/s41589-018-0004-9.
  PUBMED    29483642
REFERENCE   2  (bases 1 to 5865)
  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: "Nat Chem 
            Biol. 2018 Feb 26. pii: 10.1038/s41589-018-0004-9. doi: 
            10.1038/s41589-018-0004-9."
COMMENT     Sequence Label: pTol2-UAS-zmiRFP
FEATURES             Location/Qualifiers
     source          1..5865
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    join(14..25,26..188)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     protein_bind    704..728
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(956..980)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      988..997
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      992..1001
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    3241..3322
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3278..3297)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3332..3351
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     misc_feature    join(3712..3899,3900..3911)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(3937..3954)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4108..4696)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4188..4207)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4867..5658,5659..5727))
                     /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     5490..5509
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5728..5832)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 caggagttct tgacagaggt gtaaaaagta ctcaaaaatt ttactcaagt gaaagtacaa
       61 gtacttaggg aaaattttac tcaattaaaa gtaaaagtat ctggctagaa tcttacttga
      121 gtaaaagtaa aaaagtactc cattaaaatt gtacttgagt attaaggaag taaaagtaaa
      181 agcaagaaag aaaactagag attcttgttt aagcttttaa tctcaaaaaa cattaaatga
      241 aatgcataca aggttttatc ctgctttaga actgtttgta tttaattatc aaactataag
      301 acagacaatc taatgccagt acacgctact caaagttgta aaacctcaga tttaacttca
      361 gtagaagctg attctcaaaa ttgttagtgt caagcctagc tcttttgggg ctgaaaagca
      421 atcctgcagt gctgaaaagc ctctcacagg cagccgatgc gggaagaggt gtattagtct
      481 tgatagagag gctgcaaata gcaggaaacg tgagcagaga ctccctggtg tctgaaacac
      541 aggccagatg ggccattcgt acgcttaatt tcgaatatcg atatcaaggg cccatctcga
      601 gtatccggac acctaggtag catgcatggc gccttgtcga ctagctagca tcccgggaag
      661 aattcctcga cggatccacc ggtgtggaat tctgcagata tcaacaagtt tgtacaaaaa
      721 agcaggctgg ccatcaagct taggcctcca cggtggcttc taatccgtga gtcctagcgg
      781 gtgacagccc tccgtcttca caggcggagg agagtcttcc gtagggttcc tcggagtact
      841 gtcctccgac gcgtgcaagg gtcgactcta gagggtatat aatggatccc atcgcgtctc
      901 agcctcactt tgagctcctc cacacgaatt ccctcgacct cgaagacgcg tggataccca
      961 gctttcttgt acaaagtggt cactagtgcc accatggtgg ctggacacgc tagcggaagc
     1021 cctgacttcg gaacagctga ccctagcgac tgtgagagag aggagatcca cctggctgga
     1081 agcatccagc ctcacggaac actgctggtg gtgagcgagc ctgaccacag aatcatccag
     1141 gctagcgcta acgctgctga gttcctgaac ctgggaagcg tgctgggagt gcctctggct
     1201 gagatcgacg gagacctgct gatcaagatc ctgcctcacc tggaccctac agctgaggga
     1261 atgcctgtgg ctgtgagatg tagaatcgga aaccctagca cagagtacga cggactgatg
     1321 cacagacctc ctgagggagg actgatcatc gagctggaga gagctggacc tcctatcgac
     1381 ctgagcggaa cactggctcc tgctctggag agaatcagaa cagctggaag cctgagagct
     1441 ctgtgtgacg acacagctct gctgttccag cagtgtacag gatacgacag agtgatggtg
     1501 tacagattcg acgagcaggg acacggagag gtgtacagcg agatccacgt gacaggactg
     1561 gagagctact tcggaaacag ataccctagc agcctggtgc ctcagatggc tagaagactg
     1621 tacgagagac agagagtgag agtgctggtg gacgtgagct accagcctgt gcctctggag
     1681 cctagactga gccctctgac aggaagagac ctggacatga gcggatgttt cctgagaagc
     1741 atgagcccta cacacctgca gttcctgaag aacatgggag tgagagctac actggtggtg
     1801 agcctggtgg tgggaggaaa gctgtgggga ctggtgatct gtcaccacta cctgcctaga
     1861 ttcatccact tcgagctgag agctatctgt gagctgctgg ctgaggctat cgctacaaga
     1921 atcacagctc tgtgaggcgc gccaacccgc tcctcctccc tccctccctc cctccctcct
     1981 ctcctttatt cacttttcaa catgcacaac acgtcatcaa atcccccaat cagcactgaa
     2041 ttacagccac gggccaccag actgaaggtg ttaaagaaga cgattcggtc tcatcagcgg
     2101 ctaaaacgag atgagggtga accgagagag actgtttcca cagatgttct tgaaggaggg
     2161 ccgggctcgt aggtgcggtt attacacata taagctccca ttcacacctt aaaacatcca
     2221 acagcattcc aaagttttcc atcggggacc agggatagaa aacactgcct cttccactgc
     2281 ctaatttaac acattccttc tagattatct ctggtcatct agatgaaatc agttctgttc
     2341 tgtttgccgt gtctgaaaag acagtaaaga gtaattgaaa aaaaaaatga atatagaaaa
     2401 atggggactg gagagatatt tggactagac gctcctgatc cagccagaag gaacctttca
     2461 ggtgtcgatt acactgcctg cgaaaacaac ctcttcttcc attcttcagt ttccagctgc
     2521 cggctggctt cttccactgc ccaacatcct aacaaacccc tgacaaaaaa ggagacaagc
     2581 cttgaaacat cctgaaataa gcaattctta gctgccaggg acgtcttcct gttagcatca
     2641 gtgtatttgt ttcttttgct ttgatttgtt ttcttttttt attttatttt catggtaaat
     2701 gtattctgcc tgagcacttt aaacgaaggt gtttttgttc cagtcgatgc ttcgttttga
     2761 acctgctttt atatttatta acttttgttg tgtattttta atgtttaatc tttaaaggaa
     2821 tagttcacct acacacacac acacacacac acacacacac acaaatgaaa tgaagtcatc
     2881 atttactcac ttttctttct tttagctgtg gagcacaaaa cgagatgttt tgaagaatgt
     2941 tcacgctgtt gtttttcatg caatttaagt aaacgaagat caagggatgt tgagctatac
     3001 aaatattcat tatatgggca gcacggtcgc tggttcgagt cccggctggg ccagttggca
     3061 tttctgagtt tgcatattct ccctgtgttg gcatgggttt cctcctggtg ctccgtttct
     3121 cccacagtcc actagatcat aatcagccat accacatttg tagaggtttt acttgcttta
     3181 aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt
     3241 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     3301 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatca
     3361 ggccttaggg ataacagggt aatacgcgtg atctgcgaag atacggccac gggtgctctt
     3421 gatcctgtgg ctgattttgg actgtgctgc tcgcagctgc tgatgaatca catacttcct
     3481 ccattttctt ccactgattg actgttataa tttccctaat ttccaggtca aggtgctgtg
     3541 cattgtggta atagatgtga catgacgtca cttccaaagg accaatgaac atgtctgacc
     3601 aatttcatat aatgtgaaaa cgattttcat aggcagaata aataacattt aaattaaact
     3661 gggcatcagc gcaattcaat tggtttggta atagcaaggg aaaatagaat gaagtgatct
     3721 ccaaaaaata agtacttttt gactgtaaat aaaattgtaa ggagtaaaaa gtactttttt
     3781 ttctaaaaaa atgtaattaa gtaaaagtaa aagtattgat ttttaattgt actcaagtaa
     3841 agtaaaaatc cccaaaaata atacttaagt acagtaatca agtaaaatta ctcaagtact
     3901 ttacacctct ggttcttgag gtacctcttc cgcttcctcg ctcactgact cgctgcgctc
     3961 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     4021 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     4081 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     4141 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     4201 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     4261 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     4321 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     4381 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     4441 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     4501 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     4561 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     4621 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     4681 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     4741 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     4801 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     4861 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     4921 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     4981 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     5041 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     5101 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     5161 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     5221 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     5281 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     5341 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     5401 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     5461 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     5521 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     5581 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     5641 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     5701 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     5761 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     5821 aggggttccg cgcacatttc cccgaggtac cgatcggtat ctgtc
//
