LOCUS       sequence-145998-        3282 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Membrane targeted KOFP2 (orange) fluorophore for use with 5' 
            promoters..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3282)
  AUTHORS   Don EK, Formella I, Badrock AP, Hall TE, Morsch M, Hortle E, Hogan 
            A, Chow S, Gwee SS, Stoddart JJ, Nicholson G, Chung R, Cole NJ
  TITLE     A Tol2 Gateway-Compatible Toolbox for the Study of the Nervous 
            System and Neurodegenerative Disease.
  JOURNAL   Zebrafish. 2016 Sep 15.
  PUBMED    27631880
REFERENCE   2  (bases 1 to 3282)
  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: "Zebrafish. 
            2016 Sep 15."
COMMENT     Sequence Label: pME-mKOFP2-CAAX
FEATURES             Location/Qualifiers
     source          1..3282
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     536..553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    569..668
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     regulatory      678..687
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             684..1337
                     /codon_start=1
                     /product="monomeric Kusabira-Orange 2 fluorescent protein"
                     /label=mKO2
                     /translation="MVSVIKPEMKMRYYMDGSVNGHEFTIEGEGTGRPYEGHQEMTLRV
                     TMAEGGPMPFAFDLVSHVFCYGHRVFTKYPEEIPDYFKQAFPEGLSWERSLEFEDGGSA
                     SVSAHISLRGNTFYHKSKFTGVNFPADGPIMQNQSVDWEPSTEKITASDGVLKGDVTMY
                     LKLEGGGNHKCQMKTTYKAAKEILEMPGDHYIGHRLVRKTEGNITEQVEDAVAHS"
     protein_bind    complement(1407..1506)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1523..1542)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1524..1542)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1547..1563)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1547..1563)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1665..1684)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1676..2485
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(1743..1762)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2632..3220
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3121..3140
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac
      601 ctgttcgttg caacaaattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
      661 agcaggctga attcgccgcc accatggtga gtgtgattaa acctgagatg aagatgcgct
      721 actacatgga cggatctgtg aacggccacg agtttaccat cgagggcgag ggaaccggca
      781 gaccttacga gggacaccag gaaatgaccc tgagagtcac catggccgag ggcggaccta
      841 tgcctttcgc cttcgacctg gtgtctcacg tgttctgcta cggacacaga gtgttcacca
      901 agtaccctga ggaaatcccc gactacttca agcaggcctt ccctgaggga ctgagctggg
      961 agagaagcct ggaatttgag gacggcggaa gcgctagcgt gtccgcccac atcagcctga
     1021 gaggaaacac cttctaccac aagagcaagt tcacaggcgt gaacttccct gccgacggac
     1081 ctatcatgca gaaccagagc gtggactggg agcctagcac cgagaagatc accgccagcg
     1141 acggcgtgct gaagggcgac gtgaccatgt acctgaagct ggaaggcgga ggcaaccaca
     1201 agtgccagat gaagaccacc tacaaggccg ccaaagaaat cctggaaatg ccaggcgacc
     1261 actacatcgg acacagactg gtccgcaaga ccgagggaaa catcaccgag caggtcgagg
     1321 acgccgtggc tcacagcaag ctgaaccctc cagacgagag cggacctgga tgcatgagct
     1381 gcaaatgcgt gctctcctga actagtaccc agctttcttg tacaaagttg gcattataag
     1441 aaagcattgc ttatcaattt gttgcaacga acaggtcact atcagtcaaa ataaaatcat
     1501 tatttgccat ccagctgata tcccctatag tgagtcgtat tacatggtca tagctgtttc
     1561 ctggcagctc tggcccgtgt ctcaaaatct ctgatgttac attgcacaag ataaaaatat
     1621 atcatcatga acaataaaac tgtctgctta cataaacagt aatacaaggg gtgttatgag
     1681 ccatattcaa cgggaaacgt cgaggccgcg attaaattcc aacatggatg ctgatttata
     1741 tgggtataaa tgggctcgcg ataatgtcgg gcaatcaggt gcgacaatct atcgcttgta
     1801 tgggaagccc gatgcgccag agttgtttct gaaacatggc aaaggtagcg ttgccaatga
     1861 tgttacagat gagatggtca gactaaactg gctgacggaa tttatgcctc ttccgaccat
     1921 caagcatttt atccgtactc ctgatgatgc atggttactc accactgcga tccccggaaa
     1981 aacagcattc caggtattag aagaatatcc tgattcaggt gaaaatattg ttgatgcgct
     2041 ggcagtgttc ctgcgccggt tgcattcgat tcctgtttgt aattgtcctt ttaacagcga
     2101 tcgcgtattt cgtctcgctc aggcgcaatc acgaatgaat aacggtttgg ttgatgcgag
     2161 tgattttgat gacgagcgta atggctggcc tgttgaacaa gtctggaaag aaatgcataa
     2221 acttttgcca ttctcaccgg attcagtcgt cactcatggt gatttctcac ttgataacct
     2281 tatttttgac gaggggaaat taataggttg tattgatgtt ggacgagtcg gaatcgcaga
     2341 ccgataccag gatcttgcca tcctatggaa ctgcctcggt gagttttctc cttcattaca
     2401 gaaacggctt tttcaaaaat atggtattga taatcctgat atgaataaat tgcagtttca
     2461 tttgatgctc gatgagtttt tctaatcaga attggttaat tggttgtaac actggcagag
     2521 cattacgctg acttgacggg acggcgcaag ctcatgacca aaatccctta acgtgagtta
     2581 cgcgtcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     2641 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     2701 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     2761 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     2821 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     2881 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     2941 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     3001 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     3061 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     3121 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     3181 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     3241 tttacggttc ctggcctttt gctggccttt tgctcacatg tt
//
