LOCUS       sequence-264204-        6168 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses YFP-Sp100A fusion protein with a K297A mutation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6168)
  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 6168)
  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-Sp100A(K297A)-C1
FEATURES             Location/Qualifiers
     source          1..6168
                     /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      547..556
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(553..555,556..558,559..1269)
                     /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(598..619)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(859..878)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1206..1227
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2896..3017
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2933..2952)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2987..3006
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3024..3479)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3161..3182)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3373..3392
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3506..3610
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3612..3969
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3612..3803
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3635..3655)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3820..3955
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4004..4798
                     /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             4004..4798
                     /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(4058..4077)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4668..4687
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4986..5005)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5030..5077
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5406..5994
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5895..5914
                     /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 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
      601 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
      661 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
      721 ccctggccca ccctcgtgac caccttcggc tacggcctga agtgcttcgc ccgctacccc
      781 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
      841 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
      901 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
      961 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     1021 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     1081 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1141 cccgacaacc actacctgag ctaccagtcc aagctgagca aagaccccaa cgagaagcgc
     1201 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactcacgg catggacgag
     1261 ctgtacaagt ccggactcag atctcgagct caagcttcga attctgcagt cgacggtacc
     1321 gcgggcccgg gatctatggc aggtggaggc ggcgacctga gcaccaggag gctgaatgaa
     1381 tgtatttcac cagtagcaaa tgagatgaac catcttcctg cacacagcca cgatttgcaa
     1441 aggatgttca cggaagacca gggtgtagat gacaggttgc tctatgacat tgtattcaag
     1501 cacttcaaaa gaaataaggt ggagatttca aatgcaataa aaaagacatt tccattcctc
     1561 gagggcctcc gtgatcgtga tctcatcaca aataaaatgt ttgaagattc tcaagattct
     1621 tgtagaaacc tggtccctgt acagagagtg gtgtacaatg ttcttagtga actggagaag
     1681 acatttaacc tgccagttct ggaagcactg ttcagcgatg tcaacatgca ggaatacccc
     1741 gatttaattc acatttataa aggctttgaa aatgtaatcc atgacaaatt gcctctccaa
     1801 gaaagtgaag aagaagagag ggaggagagg tctggcctcc aactaagtct tgaacaagga
     1861 actggtgaaa actcttttcg aagcctgact tggccacctt cgggttcccc atctcatgct
     1921 ggtacaaccc cacctgaaaa tggactctca gagcacccct gtgaaacaga acagataaat
     1981 gcaaagagaa aagatacaac cagtgacaaa gatgattcgc taggaagcca acaaacaaat
     2041 gaacaatgtg ctcaaaaggc tgagccaaca gagtcctgcg aacaaattgc tgtccaagtg
     2101 aataatgggg atgctggaag ggagatgccc tgcccgttgc cctgtgatga agaaagccca
     2161 gaggcagagc tacacaacca tggaatccaa attaattcct gttctgtgcg actggtggat
     2221 ataagaaagg aaaagccatt ttctaattca agagttgagt gccaagccca agcaagaact
     2281 catcataacc aggcatctga cataatagtc atcagcagtg aggactctga aggatccact
     2341 gacgttgatg agcccttaga agtcttcatc tcagcaccga gaagcgaacc agtcattaat
     2401 aatgacaacc ctttagaatc aaatgatgaa aaggagggcc aagaagccac ttgctcacga
     2461 ccccagattg taccagagcc catggatttc agaaaattat ctacattcag agaaagtttt
     2521 aagaaaagag tgataggaca agaccacgac ttttcagaat ccagtgagga ggaggcgccc
     2581 gcagaagcct caagcggggc actgagaagc aagcatggtg agaaggctcc tatgacttct
     2641 agaagtacat ctacttggag aatacccagc aggaagagac gtttcagcag tagtgacttt
     2701 tcagacctga gtaatggaga agagcttcag gaaacctgca gctcatccct aagaagaggg
     2761 tcaggtaaag aagattaggc tagataactg atcataatca gccataccac atttgtagag
     2821 gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat
     2881 gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     2941 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     3001 ctcatcaatg tatcttaagg cgtaaattgt aagcgttaat attttgttaa aattcgcgtt
     3061 aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta
     3121 taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc
     3181 actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg
     3241 cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact
     3301 aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt
     3361 ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc
     3421 ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc
     3481 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     3541 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     3601 aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga
     3661 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     3721 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     3781 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     3841 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     3901 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     3961 ttttgcaaag atcgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg
     4021 attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca
     4081 acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt
     4141 tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg
     4201 gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga
     4261 agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca
     4321 ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct
     4381 tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac
     4441 tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc
     4501 gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt
     4561 gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt
     4621 catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg
     4681 tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat
     4741 cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc
     4801 gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc
     4861 gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc
     4921 tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc tagggggagg
     4981 ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa
     5041 agacagaata aaacgcacgg tgttgggtcg tttgttcata aacgcggggt tcggtcccag
     5101 ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc ccgcgtttct
     5161 tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc
     5221 ggggcggcag gccctgccat agcctcaggt tactcatata tactttagat tgatttaaaa
     5281 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     5341 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     5401 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     5461 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     5521 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     5581 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     5641 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     5701 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     5761 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     5821 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     5881 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     5941 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     6001 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     6061 cctgcgttat cccctgattc tgtggataac cgtattaccg ccatgcatta gttattaata
     6121 gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccg
//
