LOCUS       sequence-265263-        7368 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses YFP-Sp100C fusion protein with a W655Q mutation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7368)
  AUTHORS   Newhart A, Negorev DG, Rafalska-Metcalf IU, Yang T, Maul GG, Janicki
            SM
  TITLE     Sp100A promotes chromatin decondensation at a CMV-promoter regulated
            transcription site.
  JOURNAL   Mol Biol Cell. 2013 Mar 13.
  PUBMED    23485562
REFERENCE   2  (bases 1 to 7368)
  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: "Mol Biol 
            Cell. 2013 Mar 13."
COMMENT     Sequence Label: YFP-Sp100C(W655Q)-C3
FEATURES             Location/Qualifiers
     source          1..7368
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..204
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     155..175
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      243..252
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(249..251,252..254,255..965)
                     /codon_start=1
                     /product="enhanced YFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLKCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     primer_bind     complement(294..315)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(555..574)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     902..923
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3792..3913
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3829..3848)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3883..3902
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3920..4375)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4057..4078)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4269..4288
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4402..4506
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4508..4865
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4508..4699
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4531..4551)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4716..4851
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4900..5694
                     /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             4900..5694
                     /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(4954..4973)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5564..5583
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5882..5901)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5926..5973
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6302..6890
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6791..6810
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        7065..7368
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
       61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      121 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      181 tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta gcgctaccgg
      241 tcgccaccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg
      301 agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg
      361 ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct
      421 ggcccaccct cgtgaccacc ttcggctacg gcctgaagtg cttcgcccgc taccccgacc
      481 acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca
      541 ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg
      601 acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc
      661 tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc
      721 agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc
      781 agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg
      841 acaaccacta cctgagctac cagtccaagc tgagcaaaga ccccaacgag aagcgcgatc
      901 acatggtcct gctggagttc gtgaccgccg ccgggatcac tcacggcatg gacgagctgt
      961 acaagtactc agatctcgag ctcaagctta tggcaggtgg aggcggcgac ctgagcacca
     1021 ggaggctgaa tgaatgtatt tcaccagtag caaatgagat gaaccatctt cctgcacaca
     1081 gccacgattt gcaaaggatg ttcacggaag accagggtgt agatgacagg ttgctctatg
     1141 acattgtatt caagcacttc aaaagaaata aggtggagat ttcaaatgca ataaaaaaga
     1201 catttccatt cctcgagggc ctccgtgatc gtgatctcat cacaaataaa atgtttgaag
     1261 attctcaaga ttcttgtaga aacctggtcc ctgtacagag agtggtgtac aatgttctta
     1321 gtgaactgga gaagacattt aacctgccag ttctggaagc actgttcagc gatgtcaaca
     1381 tgcaggaata ccccgattta attcacattt ataaaggctt tgaaaatgta atccatgaca
     1441 aattgcctct ccaagaaagt gaagaagaag agagggagga gaggtctggc ctccaactaa
     1501 gtcttgaaca aggaactggt gaaaactctt ttcgaagcct gacttggcca ccttcgggtt
     1561 ccccatctca tgctggtaca accccacctg aaaatggact ctcagagcac ccctgtgaaa
     1621 cagaacagat aaatgcaaag agaaaagata caaccagtga caaagatgat tcgctaggaa
     1681 gccaacaaac aaatgaacaa tgtgctcaaa aggctgagcc aacagagtcc tgcgaacaaa
     1741 ttgctgtcca agtgaataat ggggatgctg gaagggagat gccctgcccg ttgccctgtg
     1801 atgaagaaag cccagaggca gagctacaca accatggaat ccaaattaat tcctgttctg
     1861 tgcgactggt ggatataaaa aaggaaaagc cattttctaa ttcaaaagtt gagtgccaag
     1921 cccaagcaag aactcatcat aaccaggcat ctgacataat agtcatcagc agtgaggact
     1981 ctgaaggatc cactgacgtt gatgagccct tagaagtctt catctcagca ccgagaagtg
     2041 agcctgtgat caataatgac aaccctttag aatcaaatga tgaaaaggag ggccaagaag
     2101 ccacttgctc acgaccccag attgtaccag agcccatgga tttcagaaaa ttatctacat
     2161 tcagagaaag ttttaagaaa agagtgatag gacaagacca cgacttttca gaatccagtg
     2221 aggaggaggc gcccgcagaa gcctcaagcg gggcactgag aagcaagcat ggtgagaagg
     2281 ctcctatgac ttctagaagt acatctactt ggagaatacc cagcaggaag agacgtttca
     2341 gcagtagtga cttttcagac ctgagtaatg gagaagagct tcaggaaacc tgcagctcat
     2401 ccctaagaag agggtcagga tcacagccac aagaacctga aaataagaag tgctcctgtg
     2461 tcatgtgttt tccaaaaggt gtgccaagaa gccaagaagc aaggactgaa agtagtcaag
     2521 catctgacat gatggatacc atggatgttg aaaacaattc tactttggaa aaacacagtg
     2581 ggaaaagaag aaaaaagaga aggcatagat ctaaagtaaa tggtctccaa agagggagaa
     2641 agaaagacag acctagaaaa catttaactc tgaataacaa agtccaaaag aaaagatggc
     2701 aacaaagagg aagaaaagcc aacactagac ctttgaaaag aagaagaaaa agaggtccaa
     2761 gaattcccaa agatgaaaat attaatttta aacaatctga acttcctgtg acctgtggtg
     2821 aggtgaaggg cactctatat aaggagcgat tcaaacaagg aacctcaaag aagtgtatac
     2881 agagtgagga taaaaagtgg ttcactccca gggaatttga aattgaagga gaccgcggag
     2941 catccaagaa ccagaagcta agtatacgct gcggtggata taccctgaaa gtcctgatgg
     3001 agaacaaatt tctgccagaa ccaccaagca caagaaaaaa gagaatactg gaatctcaca
     3061 acaatacctt agttgaccct tgtccggaaa actcaaatat atgtgaggtg tgcaacaaat
     3121 ggggacggct gttctgctgc gacacttgtc caagatcctt tcatgagcac tgccacatcc
     3181 catccgtgga agctaacaag aacccgtgga gttgcatctt ctgcaggata aagactattc
     3241 aggaaagatg cccagaaagc caatcaggtc atcaggaatc tgaagtcctg atgaggcaga
     3301 tgctgcctga ggagcagttg aaatgtgaat tcctcctctt gaaggtctac tgtgattcga
     3361 aaagctgctt tttcgcctca gaaccgtatt ataacagaga ggggtctcag ggcccacaga
     3421 agcccatgtg gttaaacaaa gtcaagacaa gtttgaatga gcagacgtac acccgagtag
     3481 aagggtttgt gcaggacatg cgtctcatct ttcataacca caaggaattt tacagggaag
     3541 ataaattcac cagactggga attcaagtac aggacatctt tgagaagaat ttcagaaaca
     3601 tttttgcaat tcaggaaaca agcaagaaca ttataatgtt tatttagggt accgcgggcc
     3661 cgggatccac cggatctaga taactgatca taatcagcca taccacattt gtagaggttt
     3721 tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa
     3781 ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     3841 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     3901 tcaatgtatc ttaacgcgta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat
     3961 ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa
     4021 tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta
     4081 ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca
     4141 ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat
     4201 cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg
     4261 agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc
     4321 acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt
     4381 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     4441 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     4501 aagagtcctg aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt
     4561 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     4621 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     4681 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     4741 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     4801 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     4861 gcaaagatcg atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg
     4921 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag
     4981 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt
     5041 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aagacgaggc agcgcggcta
     5101 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg
     5161 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt
     5221 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat
     5281 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg
     5341 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca
     5401 gccgaactgt tcgccaggct caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc
     5461 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc
     5521 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat
     5581 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc
     5641 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga
     5701 ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt
     5761 ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga
     5821 tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccctagg gggaggctaa
     5881 ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca ataaaaagac
     5941 agaataaaac gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg tcccagggct
     6001 ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc gtttcttcct
     6061 tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg
     6121 cggcaggccc tgccatagcc tcaggttact catatatact ttagattgat ttaaaacttc
     6181 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     6241 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     6301 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     6361 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     6421 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     6481 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     6541 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     6601 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     6661 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     6721 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     6781 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     6841 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     6901 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     6961 cgttatcccc tgattctgtg gataaccgta ttaccgccat gcattagtta ttaatagtaa
     7021 tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg
     7081 gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg
     7141 tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta
     7201 cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt
     7261 gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac
     7321 tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatg
//
