LOCUS       sequence-333606-        4669 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: N1 Cloning Vector, Excitation: 548, Emission: 561.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4669)
  AUTHORS   Karasawa S, Araki T, Nagai T, Mizuno H, Miyawaki A
  TITLE     Cyan-emitting and orange-emitting fluorescent proteins as a 
            donor/acceptor pair for fluorescence resonance energy transfer.
  JOURNAL   Biochem J. 2004 Jul 1. 381(Pt 1):307-12.
  PUBMED    15065984
REFERENCE   2  (bases 1 to 4669)
  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: "Biochem J. 
            2004 Jul 1. 381(Pt 1):307-12."
COMMENT     Sequence Label: Kusabira Orange-N1 (KO)
FEATURES             Location/Qualifiers
     source          1..4669
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        2..305
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        306..509
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     460..480
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    532..612
                     /label=MCS
                     /note="multiple cloning site"
     regulatory      614..623
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             620..1276
                     /codon_start=1
                     /product="monomeric Kusabira-Orange fluorescent protein 
                     (Karasawa et al., 2004)"
                     /label=mKO
                     /translation="MVSVIKPEMKMRYYMDGSVNGHEFTIEGEGTGRPYEGHQEMTLRV
                     TMAKGGPMPFAFDLVSHVFCYGHRPFTKYPEEIPDYFKQAFPEGLSWERSLEFEDGGSA
                     SVSAHISLRGNTFYHKSKFTGVNFPADGPIMQNQSVDWEPSTEKITASDGVLKGDVTMY
                     LKLEGGGNHKCQFKTTYKAAKKILKMPGSHYISHRLVRKTEGNITELVEDAVAHS"
     polyA_signal    1398..1519
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1435..1454)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1489..1508
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1526..1981)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1663..1684)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1875..1894
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2008..2112
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2114..2471
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2114..2305
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2137..2157)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2322..2457
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2506..3300
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             2506..3300
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2560..2579)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3170..3189
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3488..3507)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3532..3579
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3908..4496
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4397..4416
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat
       61 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc
      121 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
      181 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt
      241 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
      301 ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg
      361 gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac
      421 gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg
      481 tacggtggga ggtctatata agcagagctg gtttagtgaa ccgtcagatc cgctagcgct
      541 accggactca gatctcgagc tcaagcttcg aattctgcag tcgacggtac cgcgggcccg
      601 ggatccaccg gtcgccacca tggtgagtgt gattaaacca gagatgaaga tgaggtacta
      661 catggacggc tccgtcaatg ggcatgagtt cacaattgaa ggtgaaggca caggcagacc
      721 ttacgaggga catcaagaga tgacactacg cgtcacaatg gccaagggcg ggccaatgcc
      781 tttcgcgttt gacttagtgt cacacgtgtt ctgttacggc cacagacctt ttactaaata
      841 tccagaagag ataccagact atttcaaaca agcatttcct gaaggcctgt catgggaaag
      901 gtcgttggag ttcgaagatg gtgggtccgc ttcagtcagt gcgcatataa gccttagagg
      961 aaacaccttc taccacaaat ccaaatttac tggggttaac tttcctgccg atggtcctat
     1021 catgcaaaac caaagtgttg attgggagcc atcaaccgag aaaattactg ccagcgacgg
     1081 agttctgaag ggtgatgtta cgatgtacct aaaacttgaa ggaggcggca atcacaaatg
     1141 ccaattcaag actacttaca aggcggcaaa aaagattctt aaaatgccag gaagccatta
     1201 catcagccat cgcctcgtca ggaaaaccga aggcaacatt actgagctgg tagaagatgc
     1261 agtagctcat tcctaagcgg ccgcgactct agatcataat cagccatacc acatttgtag
     1321 aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga
     1381 atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     1441 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     1501 aactcatcaa tgtatcttaa ggcgtaaatt gtaagcgtta atattttgtt aaaattcgcg
     1561 ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct
     1621 tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt
     1681 ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat
     1741 ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca
     1801 ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac
     1861 gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta
     1921 gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg
     1981 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     2041 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     2101 aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg
     2161 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     2221 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     2281 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     2341 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     2401 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     2461 gcttttgcaa agatcgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     2521 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     2581 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     2641 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcaaga cgaggcagcg
     2701 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     2761 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     2821 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     2881 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     2941 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     3001 gcgccagccg aactgttcgc caggctcaag gcgagcatgc ccgacggcga ggatctcgtc
     3061 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     3121 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     3181 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     3241 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     3301 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt
     3361 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
     3421 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cctaggggga
     3481 ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa
     3541 aaagacagaa taaaacgcac ggtgttgggt cgtttgttca taaacgcggg gttcggtccc
     3601 agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt
     3661 cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg
     3721 tcggggcggc aggccctgcc atagcctcag gttactcata tatactttag attgatttaa
     3781 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     3841 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     3901 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     3961 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     4021 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     4081 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     4141 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     4201 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     4261 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     4321 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     4381 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     4441 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     4501 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     4561 ttcctgcgtt atcccctgat tctgtggata accgtattac cgccatgcat tagttattaa
     4621 tagtaatcaa ttacggggtc attagttcat agcccatata tggagttcc
//
