LOCUS       sequence-492857-        6046 bp    DNA     circular SYN 17-NOV-2025
DEFINITION  Localization: Intermediate Filaments, Excitation: 551, Emission: 
            565.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6046)
  TITLE     Davidson Orange Fluorescent Proteins
REFERENCE   2  (bases 1 to 6046)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Nov 17, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: mKO2-Vimentin-7
FEATURES             Location/Qualifiers
     source          1..6046
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1990..1999
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1996..2652
                     /codon_start=1
                     /product="monomeric Kusabira-Orange 2 fluorescent protein"
                     /label=mKO2
                     /translation="MVSVIKPEMKMRYYMDGSVNGHEFTIEGEGTGRPYEGHQEMTLRV
                     TMAEGGPMPFAFDLVSHVFCYGHRVFTKYPEEIPDYFKQAFPEGLSWERSLEFEDGGSA
                     SVSAHISLRGNTFYHKSKFTGVNFPADGPIMQNQSVDWEPSTEKITASDGVLKGDVTMY
                     LKLEGGGNHKCQMKTTYKAAKEILEMPGDHYIGHRLVRKTEGNITEQVEDAVAHS"
     polyA_signal    2774..2895
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2811..2830)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2865..2884
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2902..3357)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3039..3060)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3251..3270
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3384..3488
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3490..3847
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3490..3681
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3513..3533)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3698..3833
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3882..4676
                     /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             3882..4676
                     /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(3936..3955)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4546..4565
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4864..4883)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4908..4955
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5284..5872
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5773..5792
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      541 ccggactcag atctcgagct caagcttcga attcccatgt ccaccaggtc cgtgtcctcg
      601 tcctcctacc gcaggatgtt cggcggcccg ggcaccgcga gccggccgag ctccagccgg
      661 agctacgtga ctacgtccac ccgcacctac agcctgggca gcgcgctgcg ccccagcacc
      721 agccgcagcc tctacgcctc gtccccgggc ggcgtgtatg ccacgcgctc ctctgccgtg
      781 cgcctgcgga gcagcgtgcc cggggtgcgg ctcctgcagg actcggtgga cttctcgctg
      841 gccgacgcca tcaacaccga gttcaagaac acccgcacca acgagaaggt ggagctgcag
      901 gagctgaatg accgcttcgc caactacatc gacaaggtgc gcttcctgga gcagcagaat
      961 aagatcctgc tggccgagct cgagcagctc aagggccaag gcaagtcgcg cctgggggac
     1021 ctctacgagg aggagatgcg ggagctgcgc cggcaggtgg accagctaac caacgacaaa
     1081 gcccgcgtcg aggtggagcg cgacaacctg gccgaggaca tcatgcgcct ccgggagaaa
     1141 ttgcaggagg agatgcttca gagagaggaa gccgaaaaca ccctgcaatc tttcagacag
     1201 gatgttgaca atgcgtctct ggcacgtctt gaccttgaac gcaaagtgga atctttgcaa
     1261 gaagagattg cctttttgaa gaaactccac gaagaggaaa tccaggagct gcaggctcag
     1321 attcaggaac agcatgtcca aatcgatgtg gatgtttcca agcctgacct cacggctgcc
     1381 ctgcgtgacg tacgtcagca atatgaaagt gtggctgcca agaacctgca ggaggcagaa
     1441 gaatggtaca aatccaagtt tgctgacctc tctgaggctg ccaaccggaa caatgacgcc
     1501 ctgcgccagg caaagcagga gtccactgag taccggagac aggtgcagtc cctcacctgt
     1561 gaagtggatg cccttaaagg aaccaatgag tccctggaac gccagatgcg tgaaatggaa
     1621 gagaactttg ccgttgaagc tgctaactac caagacacta ttggccgcct gcaggatgag
     1681 attcagaata tgaaggagga aatggctcgt caccttcgtg aataccaaga cctgctcaat
     1741 gttaagatgg cccttgacat tgagattgcc acctacagga agctgctgga aggcgaggag
     1801 agcaggattt ctctgcctct tccaaacttt tcctccctga acctgaggga aactaatctg
     1861 gattcactcc ctctggttga tacccactca aaaaggacac ttctgattaa gacggttgaa
     1921 actagagatg gacaggttat caacgaaact tctcagcatc acgatgacct tgaaggggat
     1981 ccaccggtcg ccaccatggt gagtgtgatt aaaccagaga tgaagatgag gtactacatg
     2041 gacggctccg tcaatgggca tgagttcaca attgaaggtg aaggcacagg cagaccttac
     2101 gagggacatc aagagatgac actacgcgtc acaatggccg agggcgggcc aatgcctttc
     2161 gcgtttgact tagtgtcaca cgtgttctgt tacggccaca gagtatttac taaatatcca
     2221 gaagagatac cagactattt caaacaagca tttcctgaag gcctgtcatg ggaaaggtcg
     2281 ttggagttcg aagatggtgg gtccgcttca gtcagtgcgc atataagcct tagaggaaac
     2341 accttctacc acaaatccaa atttactggg gttaactttc ctgccgatgg tcctatcatg
     2401 caaaaccaaa gtgttgattg ggagccatca accgagaaaa ttactgccag cgacggagtt
     2461 ctgaagggtg atgttacgat gtacctaaaa cttgaaggag gcggcaatca caaatgccaa
     2521 atgaagacta cttacaaggc ggcaaaagag attcttgaaa tgccaggaga ccattacatc
     2581 ggccatcgcc tcgtcaggaa aaccgaaggc aacattactg agcaggtaga agatgcagta
     2641 gctcatagct aagcggccgc gactctagat cataatcagc cataccacat ttgtagaggt
     2701 tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc
     2761 aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     2821 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     2881 catcaatgta tcttaaggcg taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa
     2941 atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata
     3001 aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac
     3061 tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc
     3121 cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa
     3181 atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg
     3241 cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg
     3301 tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcag
     3361 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     3421 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     3481 ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa
     3541 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     3601 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     3661 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     3721 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     3781 cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     3841 ttgcaaagat cgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat
     3901 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
     3961 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
     4021 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaagacgag gcagcgcggc
     4081 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
     4141 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
     4201 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
     4261 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
     4321 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
     4381 cagccgaact gttcgccagg ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga
     4441 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     4501 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
     4561 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     4621 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg
     4681 gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga
     4741 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     4801 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccta gggggaggct
     4861 aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag
     4921 acagaataaa acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg
     4981 ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc
     5041 cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg
     5101 ggcggcaggc cctgccatag cctcaggtta ctcatatata ctttagattg atttaaaact
     5161 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     5221 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     5281 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     5341 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     5401 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     5461 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     5521 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     5581 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     5641 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     5701 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     5761 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     5821 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     5881 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     5941 tgcgttatcc cctgattctg tggataaccg tattaccgcc atgcattagt tattaatagt
     6001 aatcaattac ggggtcatta gttcatagcc catatatgga gttccg
//
