LOCUS       sequence-404997-        8076 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  This plasmid allows for inducible expression of short BRD4 isoform 
            tagged with mNeonGreen, in mammalian cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8076)
  AUTHORS   Kosno M, Currie SL, Kumar A, Xing C, Rosen MK
  TITLE     Molecular features driving condensate formation and gene expression 
            by the BRD4-NUT fusion oncoprotein are overlapping but distinct.
  JOURNAL   Sci Rep. 2023 Jul 24;13(1):11907. doi: 10.1038/s41598-023-39102-9.
  PUBMED    37488172
REFERENCE   2  (bases 1 to 8076)
  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: "Sci Rep."; 
            date: "2023-07-24"; volume: "13(1):11907. doi"; pages: " 
            10.1038/s41598-023-39102-9"
COMMENT     Sequence Label: pcDNA5-FRT/TO-mNeonGreen-BRD4
FEATURES             Location/Qualifiers
     source          1..8076
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    820..838
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    841..859
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     869..893
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             1058..1084
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     CDS             1088..1792
                     /codon_start=1
                     /product="bright monomeric yellow-green fluorescent protein
                     derived from LanYFP (Shaner et al., 2013)"
                     /label=mNeonGreen
                     /translation="VSKGEEDNMASLPATHELHIFGSINGVDFDMVGQGTGNPNDGYEE
                     LNLKSTKGDLQFSPWILVPHIGYGFHQYLPYPDGMSPFQAAMVDGSGYQVHRTMQFEDG
                     ASLTVNYRYTYEGSHIKGEAQVKGTGFPADGPVMTNSLTAADWCRSKKTYPNDKTIIST
                     FKWSYTTGNGKRYRSTARTTYTFAKPMAANYLKNQPMYVFRKTELKHSKTELNFKEWQK
                     AFTDVMGMDELYK"
     primer_bind     complement(4028..4045)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4034..4258
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4391..4410)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    4542..4589
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             4597..5619
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             4597..5619
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     polyA_signal    5749..5870
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5786..5805)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5840..5859
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5919..5935)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5919..5935)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5932..5954)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5943..5959
                     /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(5967..5973,5974..5991,5992..5997))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6012..6033
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6150..6167)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6321..6909)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6401..6420)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7080..7871,7872..7940))
                     /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     7703..7722
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7941..8045)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ccctatcagt gatagagatc
      841 tccctatcag tgatagagat cgtcgacgag ctcgtttagt gaaccgtcag atcgcctgga
      901 gacgccatcc acgctgtttt gacctccata gaagacaccg ggaccgatcc agcctccgga
      961 ctctagcgtt taaacttaag cttggtaccg agctcggatc cactagtcca gtgtggtgga
     1021 attctgcaga tatccagcac agtggcggcc gatgggaccg gctgccaagc gcgtgaaact
     1081 cgatggcgtg agcaagggcg aggaggataa catggcctct ctcccagcga cacatgagtt
     1141 acacatcttt ggctccatca acggtgtgga ctttgacatg gtgggtcagg gcaccggcaa
     1201 tccaaatgat ggttatgagg agttaaacct gaagtccacc aagggtgacc tccagttctc
     1261 cccctggatt ctggtccctc atatcgggta tggcttccat cagtacctgc cctaccctga
     1321 cgggatgtcg cctttccagg ccgcaatggt agatggctcc ggataccaag tccatcgcac
     1381 aatgcagttt gaagatggtg cctcccttac tgttaactac cgctacacct acgagggaag
     1441 ccacatcaaa ggagaggccc aggtgaaggg gactggtttc cctgctgacg gtcctgtgat
     1501 gaccaactcg ctgaccgctg cggactggtg caggtcgaag aagacttacc ccaacgacaa
     1561 aaccatcatc agtaccttta agtggagtta caccactgga aatggcaagc gctaccggag
     1621 cactgcgcgg accacctaca cctttgccaa gccaatggcg gctaactatc tgaagaacca
     1681 gccgatgtac gtgttccgta agacggaact caagcactcc aagaccgaac tcaacttcaa
     1741 ggagtggcaa aaggccttta ccgatgtgat gggcatggac gagctgtaca agggcggctc
     1801 gtacgtgtac aagtctgcgg agagcggccc tgggacgaga ttgagaaatc tgccagtaat
     1861 gggggatgga ctagaaactt cccaaatgtc tacaacacag gcccaggccc aaccccagcc
     1921 agccaacgca gccagcacca accccccgcc cccagagacc tccaacccta acaagcccaa
     1981 gaggcagacc aaccaactgc aatacctgct cagagtggtg ctcaagacac tatggaaaca
     2041 ccagtttgca tggcctttcc agcagcctgt ggatgccgtc aagctgaacc tccctgatta
     2101 ctataagatc attaaaacgc ctatggatat gggaacaata aagaagcgct tggaaaacaa
     2161 ctattactgg aatgctcagg aatgtatcca ggacttcaac actatgttta caaattgtta
     2221 catctacaac aagcctggag atgacatagt cttaatggca gaagctctgg aaaagctctt
     2281 cttgcaaaaa ataaatgagc tacccacaga agaaaccgag atcatgatag tccaggcaaa
     2341 aggaagagga cgtgggagga aagaaacagg gacagcaaaa cctggcgttt ccacggtacc
     2401 aaacacaact caagcatcga ctcctccgca gacccagacc cctcagccga atcctcctcc
     2461 tgtgcaggcc acgcctcacc ccttccctgc cgtcaccccg gacctcatcg tccagacccc
     2521 tgtcatgaca gtggtgcctc cccagccact gcagacgccc ccgccagtgc ccccccagcc
     2581 acaaccccca cccgctccag ctccccagcc cgtacagagc cacccaccca tcatcgcggc
     2641 caccccacag cctgtgaaga caaagaaggg agtgaagagg aaagcagaca ccaccacccc
     2701 caccaccatt gaccccattc acgagccacc ctcgctgccc ccggagccca agaccaccaa
     2761 gctgggccag cggcgggaga gcagccggcc tgtgaaacct ccaaagaagg acgtgcccga
     2821 ctctcagcag cacccagcac cagagaagag cagcaaggtc tcggagcagc tcaagtgctg
     2881 cagcggcatc ctcaaggaga tgtttgccaa gaagcacgcc gcctacgcct ggcccttcta
     2941 caagcctgtg gacgtggagg cactgggcct acacgactac tgtgacatca tcaagcaccc
     3001 catggacatg agcacaatca agtctaaact ggaggcccgt gagtaccgtg atgctcagga
     3061 gtttggtgct gacgtccgat tgatgttctc caactgctat aagtacaacc ctcctgacca
     3121 tgaggtggtg gccatggccc gcaagctcca ggatgtgttc gaaatgcgct ttgccaagat
     3181 gccggacgag cctgaggagc cagtggtggc cgtgtcctcc ccggcagtgc cccctcccac
     3241 caaggttgtg gccccgccct catccagcga cagcagcagc gatagctcct cggacagtga
     3301 cagttcgact gatgactctg aggaggagcg agcccagcgg ctggctgagc tccaggagca
     3361 gctcaaagcc gtgcacgagc agcttgcagc cctctctcag ccccagcaga acaaaccaaa
     3421 gaaaaaggag aaagacaaga aggaaaagaa aaaagaaaag cacaaaagga aagaggaagt
     3481 ggaagagaat aaaaaaagca aagccaagga acctcctcct aaaaagacga agaaaaataa
     3541 tagcagcaac agcaatgtga gcaagaagga gccagcgccc atgaagagca agccccctcc
     3601 cacgtatgag tcggaggaag aggacaagtg caagcctatg tcctatgagg agaagcggca
     3661 gctcagcttg gacatcaaca agctccccgg cgagaagctg ggccgcgtgg tgcacatcat
     3721 ccagtcacgg gagccctccc tgaagaattc caaccccgac gagattgaaa tcgactttga
     3781 gaccctgaag ccgtccacac tgcgtgagct ggagcgctat gtcacctcct gtttgcggaa
     3841 gaaaaggaaa cctcaagctg agaaagttga tgtgattgcc ggctcctcca agatgaaggg
     3901 cttctcgtcc tcagagtcgg agagctccag tgagtccagc tcctctgaca gcgaagactc
     3961 cgaaacaggc tcgacctgat catagtgagg ctcgagtcta gagggcccgt ttaaacccgc
     4021 tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg
     4081 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt
     4141 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc
     4201 aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct
     4261 tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc
     4321 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     4381 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     4441 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     4501 gaccccaaaa aacttgatta gggtgatggt tcacgtacct agaagttcct attccgaagt
     4561 tcctattctc tagaaagtat aggaacttcc ttggccaaaa agcctgaact caccgcgacg
     4621 tctgtcgaga agtttctgat cgaaaagttc gacagcgtct ccgacctgat gcagctctcg
     4681 gagggcgaag aatctcgtgc tttcagcttc gatgtaggag ggcgtggata tgtcctgcgg
     4741 gtaaatagct gcgccgatgg tttctacaaa gatcgttatg tttatcggca ctttgcatcg
     4801 gccgcgctcc cgattccgga agtgcttgac attggggaat tcagcgagag cctgacctat
     4861 tgcatctccc gccgtgcaca gggtgtcacg ttgcaagacc tgcctgaaac cgaactgccc
     4921 gctgttctgc agccggtcgc ggaggccatg gatgcgatcg ctgcggccga tcttagccag
     4981 acgagcgggt tcggcccatt cggaccgcaa ggaatcggtc aatacactac atggcgtgat
     5041 ttcatatgcg cgattgctga tccccatgtg tatcactggc aaactgtgat ggacgacacc
     5101 gtcagtgcgt ccgtcgcgca ggctctcgat gagctgatgc tttgggccga ggactgcccc
     5161 gaagtccggc acctcgtgca cgcggatttc ggctccaaca atgtcctgac ggacaatggc
     5221 cgcataacag cggtcattga ctggagcgag gcgatgttcg gggattccca atacgaggtc
     5281 gccaacatct tcttctggag gccgtggttg gcttgtatgg agcagcagac gcgctacttc
     5341 gagcggaggc atccggagct tgcaggatcg ccgcggctcc gggcgtatat gctccgcatt
     5401 ggtcttgacc aactctatca gagcttggtt gacggcaatt tcgatgatgc agcttgggcg
     5461 cagggtcgat gcgacgcaat cgtccgatcc ggagccggga ctgtcgggcg tacacaaatc
     5521 gcccgcagaa gcgcggccgt ctggaccgat ggctgtgtag aagtactcgc cgatagtgga
     5581 aaccgacgcc ccagcactcg tccgagggca aaggaatagc acgtactacg agatttcgat
     5641 tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg
     5701 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt
     5761 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     5821 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt
     5881 ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga
     5941 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     6001 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     6061 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     6121 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     6181 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     6241 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     6301 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     6361 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     6421 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     6481 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     6541 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     6601 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     6661 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     6721 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     6781 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     6841 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     6901 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     6961 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     7021 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     7081 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     7141 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     7201 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     7261 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     7321 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     7381 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     7441 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     7501 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     7561 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     7621 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     7681 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     7741 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     7801 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     7861 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     7921 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     7981 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     8041 ccgcgcacat ttccccgaaa agtgccacct gacgtc
//
