LOCUS       sequence-204550-        5855 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 5855)
  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 5855)
  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..5855
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    join(18..29,30..192)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     protein_bind    708..732
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(960..984)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      992..1001
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      996..1005
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    3245..3326
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3282..3301)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3336..3355
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     misc_feature    join(3716..3903,3904..3915)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(3941..3958)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4112..4700)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4192..4211)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4871..5662,5663..5731))
                     /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     5494..5513
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5732..5836)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tgtccaggag ttcttgacag aggtgtaaaa agtactcaaa aattttactc aagtgaaagt
       61 acaagtactt agggaaaatt ttactcaatt aaaagtaaaa gtatctggct agaatcttac
      121 ttgagtaaaa gtaaaaaagt actccattaa aattgtactt gagtattaag gaagtaaaag
      181 taaaagcaag aaagaaaact agagattctt gtttaagctt ttaatctcaa aaaacattaa
      241 atgaaatgca tacaaggttt tatcctgctt tagaactgtt tgtatttaat tatcaaacta
      301 taagacagac aatctaatgc cagtacacgc tactcaaagt tgtaaaacct cagatttaac
      361 ttcagtagaa gctgattctc aaaattgtta gtgtcaagcc tagctctttt ggggctgaaa
      421 agcaatcctg cagtgctgaa aagcctctca caggcagccg atgcgggaag aggtgtatta
      481 gtcttgatag agaggctgca aatagcagga aacgtgagca gagactccct ggtgtctgaa
      541 acacaggcca gatgggccat tcgtacgctt aatttcgaat atcgatatca agggcccatc
      601 tcgagtatcc ggacacctag gtagcatgca tggcgccttg tcgactagct agcatcccgg
      661 gaagaattcc tcgacggatc caccggtgtg gaattctgca gatatcaaca agtttgtaca
      721 aaaaagcagg ctggccatca agcttaggcc tccacggtgg cttctaatcc gtgagtccta
      781 gcgggtgaca gccctccgtc ttcacaggcg gaggagagtc ttccgtaggg ttcctcggag
      841 tactgtcctc cgacgcgtgc aagggtcgac tctagagggt atataatgga tcccatcgcg
      901 tctcagcctc actttgagct cctccacacg aattccctcg acctcgaaga cgcgtggata
      961 cccagctttc ttgtacaaag tggtcactag tgccaccatg gtggctggac acgctagcgg
     1021 aagccctgac ttcggaacag ctgaccctag cgactgtgag agagaggaga tccacctggc
     1081 tggaagcatc cagcctcacg gaacactgct ggtggtgagc gagcctgacc acagaatcat
     1141 ccaggctagc gctaacgctg ctgagttcct gaacctggga agcgtgctgg gagtgcctct
     1201 ggctgagatc gacggagacc tgctgatcaa gatcctgcct cacctggacc ctacagctga
     1261 gggaatgcct gtggctgtga gatgtagaat cggaaaccct agcacagagt acgacggact
     1321 gatgcacaga cctcctgagg gaggactgat catcgagctg gagagagctg gacctcctat
     1381 cgacctgagc ggaacactgg ctcctgctct ggagagaatc agaacagctg gaagcctgag
     1441 agctctgtgt gacgacacag ctctgctgtt ccagcagtgt acaggatacg acagagtgat
     1501 ggtgtacaga ttcgacgagc agggacacgg agaggtgtac agcgagatcc acgtgacagg
     1561 actggagagc tacttcggaa acagataccc tagcagcctg gtgcctcaga tggctagaag
     1621 actgtacgag agacagagag tgagagtgct ggtggacgtg agctaccagc ctgtgcctct
     1681 ggagcctaga ctgagccctc tgacaggaag agacctggac atgagcggat gtttcctgag
     1741 aagcatgagc cctacacacc tgcagttcct gaagaacatg ggagtgagag ctacactggt
     1801 ggtgagcctg gtggtgggag gaaagctgtg gggactggtg atctgtcacc actacctgcc
     1861 tagattcatc cacttcgagc tgagagctat ctgtgagctg ctggctgagg ctatcgctac
     1921 aagaatcaca gctctgtgag gcgcgccaac ccgctcctcc tccctccctc cctccctccc
     1981 tcctctcctt tattcacttt tcaacatgca caacacgtca tcaaatcccc caatcagcac
     2041 tgaattacag ccacgggcca ccagactgaa ggtgttaaag aagacgattc ggtctcatca
     2101 gcggctaaaa cgagatgagg gtgaaccgag agagactgtt tccacagatg ttcttgaagg
     2161 agggccgggc tcgtaggtgc ggttattaca catataagct cccattcaca ccttaaaaca
     2221 tccaacagca ttccaaagtt ttccatcggg gaccagggat agaaaacact gcctcttcca
     2281 ctgcctaatt taacacattc cttctagatt atctctggtc atctagatga aatcagttct
     2341 gttctgtttg ccgtgtctga aaagacagta aagagtaatt gaaaaaaaaa atgaatatag
     2401 aaaaatgggg actggagaga tatttggact agacgctcct gatccagcca gaaggaacct
     2461 ttcaggtgtc gattacactg cctgcgaaaa caacctcttc ttccattctt cagtttccag
     2521 ctgccggctg gcttcttcca ctgcccaaca tcctaacaaa cccctgacaa aaaaggagac
     2581 aagccttgaa acatcctgaa ataagcaatt cttagctgcc agggacgtct tcctgttagc
     2641 atcagtgtat ttgtttcttt tgctttgatt tgttttcttt ttttatttta ttttcatggt
     2701 aaatgtattc tgcctgagca ctttaaacga aggtgttttt gttccagtcg atgcttcgtt
     2761 ttgaacctgc ttttatattt attaactttt gttgtgtatt tttaatgttt aatctttaaa
     2821 ggaatagttc acctacacac acacacacac acacacacac acacacaaat gaaatgaagt
     2881 catcatttac tcacttttct ttcttttagc tgtggagcac aaaacgagat gttttgaaga
     2941 atgttcacgc tgttgttttt catgcaattt aagtaaacga agatcaaggg atgttgagct
     3001 atacaaatat tcattatatg ggcagcacgg tcgctggttc gagtcccggc tgggccagtt
     3061 ggcatttctg agtttgcata ttctccctgt gttggcatgg gtttcctcct ggtgctccgt
     3121 ttctcccaca gtccactaga tcataatcag ccataccaca tttgtagagg ttttacttgc
     3181 tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt
     3241 tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     3301 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     3361 atcaggcctt agggataaca gggtaatacg cgtgatctgc gaagatacgg ccacgggtgc
     3421 tcttgatcct gtggctgatt ttggactgtg ctgctcgcag ctgctgatga atcacatact
     3481 tcctccattt tcttccactg attgactgtt ataatttccc taatttccag gtcaaggtgc
     3541 tgtgcattgt ggtaatagat gtgacatgac gtcacttcca aaggaccaat gaacatgtct
     3601 gaccaatttc atataatgtg aaaacgattt tcataggcag aataaataac atttaaatta
     3661 aactgggcat cagcgcaatt caattggttt ggtaatagca agggaaaata gaatgaagtg
     3721 atctccaaaa aataagtact ttttgactgt aaataaaatt gtaaggagta aaaagtactt
     3781 ttttttctaa aaaaatgtaa ttaagtaaaa gtaaaagtat tgatttttaa ttgtactcaa
     3841 gtaaagtaaa aatccccaaa aataatactt aagtacagta atcaagtaaa attactcaag
     3901 tactttacac ctctggttct tgaggtacct cttccgcttc ctcgctcact gactcgctgc
     3961 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     4021 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     4081 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     4141 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     4201 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     4261 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     4321 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     4381 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     4441 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     4501 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat
     4561 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     4621 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     4681 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     4741 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     4801 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     4861 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     4921 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     4981 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     5041 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     5101 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     5161 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     5221 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     5281 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     5341 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     5401 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     5461 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     5521 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     5581 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     5641 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     5701 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     5761 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     5821 aaataggggt tccgcgcaca tttccccgag gtacc
//
