LOCUS       sequence-175001-        5591 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Nuclear - Nucleolus, Excitation: 548, Emission: 561.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5591)
  AUTHORS   Rizzo MA, Davidson MW, Piston DW
  TITLE     Fluorescent protein tracking and detection: fluorescent protein 
            structure and color variants.
  JOURNAL   Cold Spring Harb Protoc. 2009 Dec;2009(12):pdb.top63. doi: 
            10.1101/pdb.top63.
  PUBMED    20150100
REFERENCE   2  (bases 1 to 5591)
  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: "Cold Spring
            Harb Protoc."; date: "2009-12"; volume: "2009(12):pdb.top63. doi"; 
            pages: " 10.1101/pdb.top63"
COMMENT     Sequence Label: mKO-Fibrillarin-7
FEATURES             Location/Qualifiers
     source          1..5591
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1..654
                     /codon_start=1
                     /product="monomeric Kusabira-Orange fluorescent protein 
                     (Karasawa et al., 2004)"
                     /label=mKO
                     /translation="MVSVIKPEMKMRYYMDGSVNGHEFTIEGEGTGRPYEGHQEMTLRV
                     TMAKGGPMPFAFDLVSHVFCYGHRPFTKYPEEIPDYFKQAFPEGLSWERSLEFEDGGSA
                     SVSAHISLRGNTFYHKSKFTGVNFPADGPIMQNQSVDWEPSTEKITASDGVLKGDVTMY
                     LKLEGGGNHKCQFKTTYKAAKKILKMPGSHYISHRLVRKTEGNITELVEDAVAHS"
     CDS             complement(1450..1461)
                     /codon_start=1
                     /gene="cspA"
                     /product="translation enhancing element for E. coli (Qing 
                     et al., 2004)"
                     /label=TEE
                     /translation="NHKV"
     polyA_signal    1771..1892
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1808..1827)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1862..1881
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1899..2354)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2036..2057)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2248..2267
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2381..2485
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2487..2844
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2487..2678
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2510..2530)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2695..2830
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2879..3673
                     /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             2879..3673
                     /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(2933..2952)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3543..3562
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3861..3880)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3905..3952
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4281..4869
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4770..4789
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        5044..5347
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5348..5551
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5502..5522
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 atggtgagtg tgattaaacc agagatgaag atgaggtact acatggacgg ctccgtcaat
       61 gggcatgagt tcacaattga aggtgaaggc acaggcagac cttacgaggg acatcaagag
      121 atgacactac gcgtcacaat ggccaagggc gggccaatgc ctttcgcgtt tgacttagtg
      181 tcacacgtgt tctgttacgg ccacagacct tttactaaat atccagaaga gataccagac
      241 tatttcaaac aagcatttcc tgaaggcctg tcatgggaaa ggtcgttgga gttcgaagat
      301 ggtgggtccg cttcagtcag tgcgcatata agccttagag gaaacacctt ctaccacaaa
      361 tccaaattta ctggggttaa ctttcctgcc gatggtccta tcatgcaaaa ccaaagtgtt
      421 gattgggagc catcaaccga gaaaattact gccagcgacg gagttctgaa gggtgatgtt
      481 acgatgtacc taaaacttga aggaggcggc aatcacaaat gccaattcaa gactacttac
      541 aaggcggcaa aaaagattct taaaatgcca ggaagccatt acatcagcca tcgcctcgtc
      601 aggaaaaccg aaggcaacat tactgagctg gtagaagatg cagtagctca ttcctccgga
      661 ctcagatctc gagccatgaa gccaggattc agtccccgtg ggggtggctt tggcggccga
      721 gggggctttg gtgaccgtgg tggtcgtgga ggccgagggg gctttggcgg gggccgaggc
      781 caaggcggag gctttagagg tcgtggacga ggaggaggtg gaggcggcgg cggcggtgga
      841 ggaggaggaa gaggtggtgg aggcttccat tttggtggca accggggtcg tggtcgggga
      901 ggaaaaagag gaaaccagtc ggggaagaat gtgatggtgg agccgcatcg gcatgagggt
      961 gtcttcattt gtcggggaaa ggaagatgca ctggtcacca agaacctggt ccctggggaa
     1021 tcagtttatg gagagaagag agtctcgatt tcggaaggag atgacaaaat tgagtaccga
     1081 gcctggaacc ccttccgctc caagctagca gcagcaatcc tgggtggtgt ggaccagatc
     1141 cacatcaaac cgggggctaa ggttctctac ctcggggctg cctcgggcac cacggtctcc
     1201 catgtctctg acatcgttgg tccggatggt ctagtctatg cagtcgagtt ctcccaccgc
     1261 tctggccgtg acctcattaa cttggccaag aagaggacca acatcattcc tgtgatcgag
     1321 gatgctcgac acccacacaa ataccgcatg ctcatcgcaa tggtggatgt gatctttgct
     1381 gatgtggccc agccagacca gacccggatt gtggccctga atgcccacac cttcctgcgt
     1441 aatggaggac actttgtgat ttccattaag gccaactgca ttgactccac agcctcagcc
     1501 gaggccgtgt ttgcctccga agtgaaaaag atgcaacagg agaacatgaa gccgcaggag
     1561 cagttgaccc ttgagccata tgaaagagac catgccgtgg tcgtgggagt gtacaggcca
     1621 ccccccaagg tgaagaactg aggatccacc ggatctagat aactgatcat aatcagccat
     1681 accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg
     1741 aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac
     1801 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     1861 tgtggtttgt ccaaactcat caatgtatct taacgcgtaa attgtaagcg ttaatatttt
     1921 gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat
     1981 cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt
     2041 ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt
     2101 ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag
     2161 gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg
     2221 aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc
     2281 gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc
     2341 gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     2401 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     2461 tcaataatat tgaaaaagga agagtcctga ggcggaaaga accagctgtg gaatgtgtgt
     2521 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     2581 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     2641 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     2701 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     2761 tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt
     2821 tttggaggcc taggcttttg caaagatcga tcaagagaca ggatgaggat cgtttcgcat
     2881 gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg
     2941 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc
     3001 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca
     3061 agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct
     3121 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga
     3181 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg
     3241 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat
     3301 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga
     3361 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg
     3421 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg
     3481 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat
     3541 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct
     3601 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga
     3661 cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg
     3721 ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt
     3781 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc
     3841 caccctaggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct
     3901 atgacggcaa taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc
     3961 ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa
     4021 tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg
     4081 ctcgcagcca acgtcggggc ggcaggccct gccatagcct caggttactc atatatactt
     4141 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     4201 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     4261 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     4321 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     4381 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     4441 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     4501 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     4561 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     4621 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     4681 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     4741 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     4801 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     4861 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     4921 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccatg
     4981 cattagttat taatagtaat caattacggg gtcattagtt catagcccat atatggagtt
     5041 ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc
     5101 attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg
     5161 tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat
     5221 gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca
     5281 gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat
     5341 taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg
     5401 gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca
     5461 acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg
     5521 tgtacggtgg gaggtctata taagcagagc tggtttagtg aaccgtcaga tccgctagcg
     5581 ctaccggtac c
//
