LOCUS       sequence-382119-        7292 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses nuclear-localized FR-MQV in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7292)
  AUTHORS   Mukherjee S, Hung ST, Douglas N, Manna P, Thomas C, Ekrem A, Palmer 
            AE, Jimenez R
  TITLE     Engineering of a Brighter Variant of the FusionRed Fluorescent 
            Protein Using Lifetime Flow Cytometry and Structure-Guided 
            Mutations.
  JOURNAL   Biochemistry. 2020 Oct 6;59(39):3669-3682. doi: 
            10.1021/acs.biochem.0c00484. Epub 2020 Sep 24.
  PUBMED    32914619
REFERENCE   2  (bases 1 to 7292)
  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"; doi: 
            "10.1021/acs.biochem.0c00484"; journalName: "Biochemistry"; date: 
            "2020-10-6- 6"; volume: "59"; issue: "39"; pages: "3669-3682"
COMMENT     Sequence Label: piggyBAC-H2B FR-MQV
FEATURES             Location/Qualifiers
     source          1..7292
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(1..105)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(131..586)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(268..289)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     480..499
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     713..735
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     727..744
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     728..744
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   874..936
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     promoter        1440..1643
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1594..1614
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1640..1664
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             1779..2156
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAITKYTSAK"
     CDS             2169..2867
                     /codon_start=1
                     /product="FusionRed fluorescent protein derivative that 
                     shows reversible photoswitching with visible light"
                     /label=rsFusionRed1
                     /translation="MVSELIKENMPMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTQRIK
                     VVEGGPLPFAFDILATSFMYGSRTFIKHPPGIPDFFKQSFPEGFTWERVTTYEDGGVLT
                     ATQDTSLQDGCLIYNVKVRGVNFPANGPVMQKKTLGWEASTETMYPADGGLEGAVDMAL
                     KLVGGGHLICNMETTYRSKKPATNLKMPGVYNVDHRLERIKEADDETYVEQHEVAVARY
                     STGGAGDGGK"
     CDS             3141..3740
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(3141..3160)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3637..3657
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(3800..3820)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    3862..3983
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3899..3918)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3953..3972
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   4746..4780
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(5271..5287)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5271..5287)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5284..5306)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5295..5311
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(5319..5325,5326..5343,5344..5349))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5364..5385
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5502..5519)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5673..6261)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5753..5772)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6432..7223,7224..7292))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7055..7074
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
       61 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
      121 agtgccacct aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
      181 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
      241 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac
      301 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa
      361 ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct
      421 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa
      481 gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc
      541 gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtccca ttcgccattc
      601 aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg
      661 gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca
      721 cgacgttgta aaacgacggc cagtgagcgc gcctcgttca ttcacgtttt tgaacccgtg
      781 gaggacgggc agactcgcgg tgcaaatgtg ttttacagcg tgatggagca gatgaagatg
      841 ctcgacacgc tgcagaacac gcagctagat taaccctaga aagataatca tattgtgacg
      901 tacgttaaag ataatcatgt gtaaaattga cgcatgtgtt ttatcggtct gtatatcgag
      961 gtttatttat taatttgaat agatattaag ttttattata tttacactta catactaata
     1021 ataaattcaa caaacaattt atttatgttt atttatttat taaaaaaaac aaaaactcaa
     1081 aatttcttct ataaagtaac aaaactttta tgagggacag ccccccccca aagcccccag
     1141 ggatgtaatt acgtccctcc cccgctaggg ggcagcagcg agccgcccgg ggctccgctc
     1201 cggtccggcg ctccccccgc atccccgagc cggcagcgtg cggggacagc ccgggcacgg
     1261 ggaaggtggc acgggatcgc tttcctctga acgcttctcg ctgctctttg agcctgcaga
     1321 cacctggggg gatacgggga aaaggcctcc acggccacta gtattatgcc cagtacatga
     1381 ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg
     1441 tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc
     1501 caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact
     1561 ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt
     1621 gggaggttta tataagcaga gctcgtttag tgaaccgtca gatcgcctgg agacgccatc
     1681 cacgctgttt tgacctccat agaagattct agagctagcg aattcgctac cggtctcgag
     1741 ctcaagcttc gaattctgca gtcgacggta ccgccaccat gccagagcca gcgaagtctg
     1801 ctcccgcccc gaaaaagggc tccaagaagg cggtgactaa ggcgcagaag aaaggcggca
     1861 agaagcgcaa gcgcagccgc aaggagagct attccatcta tgtgtacaag gttctgaagc
     1921 aggtccaccc tgacaccggc atttcgtcca aggccatggg catcatgaat tcgtttgtga
     1981 acgacatttt cgagcgcatc gcaggtgagg cttcccgcct ggcgcattac aacaagcgct
     2041 cgaccatcac ctccagggag atccagacgg ccgtgcgcct gctgctgcct ggggagttgg
     2101 ccaagcacgc cgtgtccgag ggtactaagg ccatcaccaa gtacaccagc gctaaggatc
     2161 ccaaaaaaat ggtgtccgag ctgattaagg agaacatgcc tatgaaactg tatatggagg
     2221 gaaccgtcaa caaccaccac ttcaaatgca caagcgaggg cgaagggaag ccttacgagg
     2281 ggacacagac tcagaggatc aaagtggtcg aaggcgggcc actgcccttc gcatttgaca
     2341 ttctggccac cagcttcatg tatgggtccc gcacttttat caagcacccc cctggcatcc
     2401 ccgacttctt taaacagtct ttccccgagg gcttcacctg ggaacgagtg accacatacg
     2461 aggatggagg cgtcctgacc gccacacagg acacaagtct ccaggatggc tgcctgatct
     2521 ataacgtgaa ggtcagagga gtgaacttcc ctgccaatgg cccagtcatg cagaagaaaa
     2581 ccctgggctg ggaggccagc actgaaacca tgtacccagc agacggagga ctggagggag
     2641 cagtcgatat ggctctgaaa ctggtgggcg ggggacacct gatctgtaat atggaaacta
     2701 cctaccggag caagaagccc gccactaacc tgaagatgcc tggcgtgtat aatgtcgatc
     2761 atcggctgga gcggatcaag gaagctgacg atgagacata tgtggaacag cacgaagtgg
     2821 cagtggcaag atactcaaca gggggggcag gcgatggagg gaaatgagcg gccgccgccc
     2881 tacctgaggc cgccatccac gccggttgag tcgcgttctg ccgcctcccg cctgtggtgc
     2941 ctcctgaact gcgtccgccg tctaggtaag tttaaagctc aggtcgagac cgggcctttg
     3001 tccggcgctc ccttggagcc tacctagact cagccggctc tccacgcttt gcctgaccct
     3061 gcttgctcaa ctctacgtct ttgtttcgtt ttctgttctg cgccgttaca gatccaagct
     3121 gtgaccggcg cctacgctag atgaccgagt acaagcccac ggtgcgcctc gccacccgcg
     3181 acgacgtccc cagggccgta cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc
     3241 gccacaccgt cgatccggac cgccacatcg agcgggtcac cgagctgcaa gaactcttcc
     3301 tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg
     3361 cggtctggac cacgccggag agcgtcgaag cgggggcggt gttcgccgag atcggcccgc
     3421 gcatggccga gttgagcggt tcccggctgg ccgcgcagca acagatggaa ggcctcctgg
     3481 cgccgcaccg gcccaaggag cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc
     3541 accagggcaa gggtctgggc agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg
     3601 ccggggtgcc cgccttcctg gagacctccg cgccccgcaa cctccccttc tacgagcggc
     3661 tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga
     3721 cccgcaagcc cggtgcctga gtcgacaatc aacctctgga ttacaaaatt tgtgaaagat
     3781 tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc
     3841 ctttgtatca tgcgttaact aaacttgttt attgcagctt ataatggtta caaataaagc
     3901 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     3961 tccaaactca tcaatgtatc ttatcatgtc tggaattgac tcaaatgatg tcaattagtc
     4021 tatcagaagc tcatctggtc tcccttccgg gggacaagac atccctgttt aatatttaaa
     4081 cagcagtgtt cccaaactgg gttcttatat cccttgctct ggtcaaccag gttgcagggt
     4141 ttcctgtcct cacaggaacg aagtccctaa agaaacagtg gcagccaggt ttagccccgg
     4201 aattgactgg attccttttt tagggcccat tggtatggct ttttccccgt atccccccag
     4261 gtgtctgcag gctcaaagag cagcgagaag cgttcagagg aaagcgatcc cgtgccacct
     4321 tccccgtgcc cgggctgtcc ccgcacgctg ccggctcggg gatgcggggg gagcgccgga
     4381 ccggagcgga gccccgggcg gctcgctgct gccccctagc gggggaggga cgtaattaca
     4441 tccctggggg ctttgggggg gggctgtccc tgatatctat aacaagaaaa tatatatata
     4501 ataagttatc acgtaagtag aacatgaaat aacaatataa ttatcgtatg agttaaatct
     4561 taaaagtcac gtaaaagata atcatgcgtc attttgactc acgcggtcgt tatagttcaa
     4621 aatcagtgac acttaccgca ttgacaagca cgcctcacgg gagctccaag cggcgactga
     4681 gatgtcctaa atgcacagcg acggattcgc gctatttaga aagagagagc aatatttcaa
     4741 gaatgcatgc gtcaatttta cgcagactat ctttctaggg ttaatctagc tgcatcagga
     4801 tcatatcgtc gggtcttttt tccggctcag tcatcgccca agctggcgct atctgggcat
     4861 cggggaggaa gaagcccgtg ccttttcccg cgaggttgaa gcggcatgga aagagtttgc
     4921 cgaggatgac tgctgctgca ttgacgttga gcgaaaacgc acgtttacca tgatgattcg
     4981 ggaaggtgtg gccatgcacg cctttaacgg tgaactgttc gttcaggcca cctgggatac
     5041 cagttcgtcg cggcttttcc ggacacagtt ccggatggtc agcccgaagc gcatcagcaa
     5101 cccgaacaat accggcgaca gccggaactg ccgtgccggt gtgcagatta atgacagcgg
     5161 tgcggcgctg ggatattacg tcagcgagga cgggtatcct ggctggatgc cgcagaaatg
     5221 gacatggata ccccgtgagt tacccggcgg gcgcgcttgg cgtaatcatg gtcatagctg
     5281 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     5341 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     5401 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     5461 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     5521 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     5581 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     5641 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     5701 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     5761 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     5821 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     5881 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     5941 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     6001 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     6061 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     6121 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     6181 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     6241 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     6301 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     6361 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     6421 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     6481 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     6541 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     6601 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     6661 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     6721 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     6781 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     6841 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     6901 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     6961 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     7021 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     7081 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     7141 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     7201 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     7261 ataagggcga cacggaaatg ttgaatactc at
//
