LOCUS       sequence-324822-        6586 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Acute Depletion of BRM-EYFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6586)
  AUTHORS   Zhu Y, Denholtz M, Lu H, Murre C
  TITLE     Calcium signaling instructs NIPBL recruitment at active enhancers 
            and promoters via distinct mechanisms to reconstruct genome 
            compartmentalization.
  JOURNAL   Genes Dev. 2021 Jan 1;35(1-2):65-81. doi: 10.1101/gad.343475.120. 
            Epub 2020 Dec 17.
  PUBMED    33334824
REFERENCE   2  (bases 1 to 6586)
  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: "Genes 
            Dev."; date: "2021-01-1"; volume: "35(1-2):65-81. doi"; pages: " 
            10.1101/gad.343475.120. Epub 2020 Dec 17"
COMMENT     Sequence Label: pFlox_N-BRM_BSD_FKBPF36V_EYFPF36V
FEATURES             Location/Qualifiers
     source          1..6586
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     127..149
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             1513..1908
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     CDS             1918..1974
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             1975..1998
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2008..2328
                     /codon_start=1
                     /product="F36V mutant of FK506-binding protein FKBP12"
                     /label=DmrB
                     /note="binds synthetic ligands that do not bind wild-type 
                     FKBP"
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKVDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     CDS             2335..2361
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             join(2362..2364,2365..3075)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(2404..2425)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2665..2684)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3012..3033
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     promoter        complement(4089..4107)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4128..4144)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4128..4144)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4141..4163)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4152..4168
                     /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(4176..4182,4183..4200,4201..4206))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4221..4242
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4359..4376)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4530..5118)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4610..4629)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5289..6080,6081..6149))
                     /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     5912..5931
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6150..6254)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6322..6340
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6378..6400)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6500..6519
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc
       61 tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag
      121 ggttttccca gtcacgacgt tgtaaaacga cggccgcggc ggccgcacta gtggccgcgg
      181 cggccgcact agtggccgcg gcggccgcag cttgcctgca ggtcgacgcc aggaagctta
      241 gaggctgggt gatgggggga gtgttcagtt aagtctggag aaacagaacg cccattcgca
      301 gacacagaca gctacaccct ggcatccttg accagaggcc tagcacgaat catgagtctt
      361 agttgtccag gttttggttc taatacttaa cctaggagga ggaccccagc tttgtccatt
      421 ttgataactt gaggtccagt agctgtggtg ggtaggtgag gcaggcagct gtggttggaa
      481 ttctgctgtt gttttacatg tatgtctcat ctgactctaa gaagtatcaa gacaagaact
      541 tatctaataa ttgtgtctga atctatttga gcctatattg tatgatccct gtagcttgct
      601 tgctctctct ctctctctcc ctctctctct ctcccctttc tctctccccc tccctctccc
      661 cctctccttc cctctctccc tctccccctt cccctcccct tttcccctcc ccttccctct
      721 cccctcccac atttcctctc cctctctctg ttttgttgtt gttttgtttt tgaggcaggg
      781 tttctctgtg tagtgctggt tgtcctggaa cttgctctgt ccaccaggct ggcttggacc
      841 tcacagagct cctcctgctt ctgcctccca agtgctagga ggcacctatg tgccaccctg
      901 ttgttatttt tcttcctttt ctataattga gacttctctt tttattatca attgatggac
      961 ttatttttcc cccttttagc tatctagaaa tcttggtgaa gttttgattg cctgttggct
     1021 tgtgaaaatg agtcttggtt ggatacttat ttgcaaatgg ttttccagtc cagtggtttg
     1081 ggggagtggg ggcttaatga cttctgaggt gtctaagcca gcctataaca gagcgtatct
     1141 gttgtgttta gtagtaaatt tcagacttgt ttagattctg cttatgtgtt aaatttcagt
     1201 tgcttcagtg agtgtctcta tatccagggt gatttatcat ttggggggaa tcacttaaag
     1261 tatgtgtttt ttaaaacaag tggtcttcat aaaatctgag cccagggtta tatatcacat
     1321 tcattcaaac atctggaaga ggccagggag ttctgtttga tgttagttac agaaatggca
     1381 ccatcagatt ttaacaactg ttttatttgg accttaacat catgtttaaa acatttcttt
     1441 ctttcttttt cttcctgctc tccaacttag tgtcactctg ctaactggca gagccaggag
     1501 acctaggtcg acatggccaa gcctttgtct caagaagaat ccaccctcat tgaaagagca
     1561 acggctacaa tcaacagcat ccccatctct gaagactaca gcgtcgccag cgcagctctc
     1621 tctagcgacg gccgcatctt cactggtgtc aatgtatatc attttactgg gggaccttgt
     1681 gcagaactcg tggtgctggg cactgctgct gctgcggcag ctggcaacct gacttgtatc
     1741 gtcgcgatcg gaaatgagaa caggggcatc ttgagcccct gcggacggtg ccgacaggtg
     1801 cttctcgatc tgcatcctgg gatcaaagcc atagtgaagg acagtgatgg acagccgacg
     1861 gcagttggga ttcgtgaatt gctgccctct ggttatgtgt gggagggcgg ttccggagcc
     1921 acgaacttct ctctgttaaa gcaagcagga gacgtggaag aaaaccccgg tcccgattac
     1981 aaggatgacg atgacaaggg ttccggcggc gtgcaggtgg agactatctc cccaggagac
     2041 gggcgcacct tccccaagcg cggccagacc tgcgtggtgc actacaccgg gatgcttgaa
     2101 gatggaaaga aagttgattc ctcccgggac agaaacaagc cctttaagtt tatgctaggc
     2161 aagcaggagg tgatccgagg ctgggaagaa ggggttgccc agatgagtgt gggtcagaga
     2221 gccaaactga ctatatctcc agattatgcc tatggtgcca ctgggcaccc aggcatcatc
     2281 ccaccacatg ccactctcgt cttcgatgtg gagcttctaa aactggaagt cgattatccg
     2341 tacgacgtac cagactacgc agtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc
     2401 atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc
     2461 gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg
     2521 cccgtgccct ggcccaccct cgtgaccacc ttcggctacg gcctgcagtg cttcgcccgc
     2581 taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc
     2641 caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag
     2701 ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac
     2761 ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg
     2821 gccgacaagc agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac
     2881 ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg
     2941 ctgctgcccg acaaccacta cctgagctac cagtccgccc tgagcaaaga ccccaacgag
     3001 aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg
     3061 gacgagctgt acaagggtgg aggtggatct ggtggcggcg gttcaggtac ctccacaccc
     3121 acagacccag cagcaatgcc ccatcctggg ccctccccgg ggcctggacc ctctcctgga
     3181 ccaattctgg ggcctagtcc aggaccagga ccatccccag gttctgtgca cagcatgatg
     3241 ggtcctagtc ccggacctcc cagcgtctca catcctctgt caacgatggg ctctgcagac
     3301 ttcccacagg aaggcatgca ccaattacat aaggtaagag tctgtgccct ctcccccact
     3361 ttacccgtct tctcaggcat tctgatgact ctgagctttc tcctgctagt aacataagaa
     3421 tctagctaga aatttgtgtt aacttgcatg ttattaaaaa cattcctgta atcccagcac
     3481 ttgagagact gaagcaaaag gatcatgcat taaagaccaa tctggatctt ataaaaaccc
     3541 aaaccaaacc acaaccaagc ccaatataaa gcaaaaacca tccccaaact ggggtaaaac
     3601 acttctactg ctgtattttt tactgttacg gatggcccag gaatcgagaa gttgtctttt
     3661 caaacacaaa ggtccgccat gcaatttgga caggtttggg gattggtggg ttaaaggtag
     3721 tttgtcttgg tgaacgtgta tctcagtaag ggagacttcc ttccctcacc tcccatgtac
     3781 tttatatcac tcacaaatca atatattaga aattagtcct ccctacaaat gttgaaattt
     3841 atttgatgaa actagatctg taagagtgaa ggtgaaagtt gccttctgaa ggtattgcag
     3901 ggcctggtga gagagagaga gagagagaga gagagagaga gagagagaga gagagagatc
     3961 agcacctaag agcatgtgaa gcactaccat tgggcagctc acactggggt accctcgagg
     4021 tcgatcgacg gtatcgataa gcttgatcta gagcggccgc caccgcggtg gagctccagc
     4081 ttttgttccc tttagtgagg gttaatttcg agcttggcgt aatcatggtc atagctgttt
     4141 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     4201 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     4261 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     4321 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     4381 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     4441 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     4501 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     4561 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     4621 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     4681 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     4741 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     4801 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     4861 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     4921 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     4981 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     5041 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     5101 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     5161 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     5221 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     5281 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     5341 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     5401 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     5461 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     5521 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     5581 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     5641 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     5701 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     5761 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     5821 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     5881 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     5941 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     6001 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     6061 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     6121 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     6181 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     6241 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt
     6301 atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc
     6361 ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg
     6421 taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt
     6481 cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatgcgg
     6541 tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggc
//
