LOCUS       sequence-323487-        6522 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Acute Depletion of BRG1-mScarlet.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6522)
  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 6522)
  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-BRG1_Neo_FKBPF36V_mScarlet
FEATURES             Location/Qualifiers
     source          1..6522
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     127..149
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             876..1667
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(930..949)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1540..1559
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             1677..1733
                     /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             1734..1760
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YPYDVPDYA"
     CDS             1734..1760
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1764..1790
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1800..2120
                     /codon_start=1
                     /product="F36V mutant of FK506-binding protein FKBP12"
                     /label=FKBP-V
                     /note="binds synthetic ligands that do not bind wild-type 
                     FKBP"
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKVDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     CDS             2151..2846
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet-I
                     /note="matures faster than mScarlet"
                     /translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFSWDILSPQFMYGSRAFIKHPADIPDYYKQSFPEGFKWERVMNFEDGG
                     AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIK
                     MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH
                     STGGMDELYK"
     primer_bind     complement(2284..2302)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2552..2571
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     promoter        complement(4025..4043)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4064..4080)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4064..4080)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4077..4099)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4088..4104
                     /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(4112..4118,4119..4136,4137..4142))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4157..4178
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4295..4312)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4466..5054)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4546..4565)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5225..6016,6017..6085))
                     /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     5848..5867
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6086..6190)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6258..6276
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6314..6336)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6436..6455
                     /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 ggtcgacgat tagcactcac
      241 tgattgtttt tgtgtgtctg tctgactgcc tgtcttgttc atgaccttca tgaacaagag
      301 aacagatgac tccacagtta aaggataatt tgagagttgc cccacagtct gctggggacc
      361 tagttgatct tggaggtaat gggaggacat tgtcccactg accacatcaa gtgtagcttc
      421 ctagagtgat tattggtcag tttccccagt ctgtcacttt atagttagat aaggctgggc
      481 aatacgcaga agtaattttg tggtagggtg gggagtgaga cagggtctcc ctatgcagtc
      541 ctggctggcc tggaactagt atagaactca aattaagaga tcctctgcct ccctatttct
      601 ggggtcatgg acatgcacca tcattccctg ctttctgagc tgatttcttt tcctgagata
      661 acattgtttt tcccaaaaag aataaaagaa aggtttggaa caaaggttag gtatcaggcc
      721 tacaagatgg ggctcagtgg tctgggtgaa cctgactgac ctttgggccc ttctttgctg
      781 ttcttgacat aactgaaccc ctgtcccaga ttctaggatg accagaattt ctttccaaca
      841 ggagaccagt gcctgtagct ccagtaaagg tcgacatgat tgaacaagat ggattgcacg
      901 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa
      961 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg
     1021 tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt
     1081 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa
     1141 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc
     1201 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
     1261 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
     1321 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
     1381 aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg
     1441 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
     1501 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg
     1561 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
     1621 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttcggt tccggagcta
     1681 ctaacttcag cctgctgaag caggctggag acgtggagga gaaccctgga ccttacccct
     1741 acgacgtgcc cgactacgcc ggctacccat acgatgttcc agattacgct ggaagcggag
     1801 gagtgcaggt ggaaaccatc tccccaggag acgggcgcac cttccccaag cgcggccaga
     1861 cctgcgtggt gcactacacc gggatgcttg aagatggaaa gaaagttgat tcctcccggg
     1921 acagaaacaa gccctttaag tttatgctag gcaagcagga ggtgatccga ggctgggaag
     1981 aaggggttgc ccagatgagt gtgggtcaga gagccaaact gactatatct ccagattatg
     2041 cctatggtgc cactgggcac ccaggcatca tcccaccaca tgccactctc gtcttcgatg
     2101 tggagcttct aaaactggaa ggtggaggcg ggagtggagg tggaggatcg atggtgagca
     2161 agggcgaggc agtgatcaag gagttcatgc ggttcaaggt gcacatggag ggctccatga
     2221 acggccacga gttcgagatc gagggcgagg gcgagggccg cccctacgag ggcacccaga
     2281 ccgccaagct gaaggtgacc aagggtggcc ccctgccctt ctcctgggac atcctgtccc
     2341 ctcagttcat gtacggctcc agggccttca tcaagcaccc cgccgacatc cccgactact
     2401 ataagcagtc cttccccgag ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg
     2461 gcgccgtgac cgtgacccag gacacctccc tggaggacgg caccctgatc tacaaggtga
     2521 agctccgcgg caccaacttc cctcctgacg gccccgtaat gcagaagaag acaatgggct
     2581 gggaagcgtc caccgagcgg ttgtaccccg aggacggcgt gctgaagggc gacattaaga
     2641 tggccctgcg cctgaaggac ggcggccgct acctggcgga cttcaagacc acctacaagg
     2701 ccaagaagcc cgtgcagatg cccggcgcct acaacgtgga ccgcaagttg gacatcacct
     2761 cccacaacga ggactacacc gtggtggaac agtacgaacg ctccgagggc cgccactcca
     2821 ccggcggcat ggacgagctg tacaagggtg gcggtggctc gggcggtggt gggtcgggtg
     2881 gcggcggatc tggtaccttg ggtgggactc ctcggcctgg tccttcccca ggccctggtc
     2941 cttcacctgg tgcaatgctg ggtcctagcc ctggcccctc accaggttct gcccacagca
     3001 tgatggggcc aagcccagga cctccttcag caggacatcc catgcccacc caggggcctg
     3061 gagggtaccc ccaggacaac atgcatcaga tgcacaaggt aggcatcctg tgatctcagt
     3121 cctgtactct gttcctgaga cttgccttgg tgtctcatac aaagctgcca gagcctccgg
     3181 acattagagg agagaggtct gtggaggtga cttggcttag ttcagggtcc taccctaaag
     3241 tgtgcttctt tgaacaaagg aatcaggctg ttgtcctgag ccaggaggct gagcagccat
     3301 ttcactctac gtttcccatt ctccctatgc tttcttttgt taaaaggaat tcccagattc
     3361 ctctaaggac gatcctctgg ctgcactggt ggcctacata cttgaagcag tggggagtat
     3421 tgagtctgct tctgagcttg taccccagac tgccatcttc ctttggtaaa tgtgtcagca
     3481 ttgcttttgc cagcaaggct gccaggcagc aatcctgtcc ttaggccttg gtgttcctct
     3541 taggactggc gcagtagaat gtcagcattt atctgggctg acccacagag gctggggtgt
     3601 aagaagatat tggaggctgc aggagggttc accatagtgg cattgtcagt tgctcctctt
     3661 ccctgcaggg ttgtcctctg tttgcttcct aatcctgcaa gtataggttg tgttccctat
     3721 ccacaaatga ggaaggccct cacatgcctg tctcattcct cggcctggct tgtccttgta
     3781 ctctgcttcc agacatctcc atctggctcc ccagttctgg atggttgtgt catcttagta
     3841 gagtacttat cttgctcttt cccctatgaa ggaagctctg ggtcccatgt aggagcccac
     3901 cgtctggttt acagggccct ccatgtccgg taagctgcat ctcggtaccc tcgaggtcga
     3961 tcgacggtat cgataagctt gatctagagc ggccgccacc gcggtggagc tccagctttt
     4021 gttcccttta gtgagggtta atttcgagct tggcgtaatc atggtcatag ctgtttcctg
     4081 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     4141 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     4201 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     4261 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     4321 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     4381 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     4441 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     4501 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     4561 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     4621 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     4681 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     4741 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     4801 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     4861 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     4921 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     4981 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     5041 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     5101 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     5161 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     5221 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     5281 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     5341 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     5401 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     5461 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     5521 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     5581 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     5641 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     5701 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     5761 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     5821 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     5881 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     5941 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     6001 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     6061 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     6121 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     6181 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca
     6241 tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg
     6301 atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag
     6361 cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg
     6421 gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg
     6481 aaataccgca cagatgcgta aggagaaaat accgcatcag gc
//
