LOCUS       sequence-323492-        5897 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Acute Depletion of RD21-EYFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5897)
  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 5897)
  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 c-RAD21_FKBPF36V_EYFP_Bsd
FEATURES             Location/Qualifiers
     source          1..5897
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     127..149
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             1107..1130
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1140..1460
                     /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             1467..1493
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             join(1494..1496,1497..2207)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(1536..1557)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1797..1816)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2144..2165
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2217..2273
                     /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             2274..2672
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     promoter        complement(3400..3418)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3439..3455)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3439..3455)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3452..3474)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3463..3479
                     /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(3487..3493,3494..3511,3512..3517))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3532..3553
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3670..3687)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3841..4429)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3921..3940)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4600..5391,5392..5460))
                     /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     5223..5242
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5461..5565)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5633..5651
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5689..5711)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5811..5830
                     /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 ggtcgacagt cctttcccag
      241 aaagcaagtc ctgttaaacc ccgaagcatg ccttggtggg aatttctcca gggctgcatt
      301 gttttaaagc atcttagtta catacatgaa gtgtcaagcc aggggtaaac ctagagcatt
      361 tagtttatgt agccatgatt atcctaaaaa acctgatgca tggtgctgtg atggggagcg
      421 cagctgcgta ctgtcttgtt tgattagcaa aagccatgtg ctaagtcata gagtagcaga
      481 agtcagcaac ctcagctagg gttccagata caggtgaggc tggagcaccc gtgacagttc
      541 tcccaagttc cttcctttct gcacatgggg atggatgata cttagggttg gaaggttatc
      601 aggggccggt ttgattttgg ttttgttttc actttaaaat ctgctgagtg tttgtttttg
      661 ctaactcaca tctctgttgt ggcgccttca cttacttctt gaactctgta tgccttaaga
      721 taatatgctt tagataataa ggcctttacc ttagtgacta gcatgggaac cacttgcgta
      781 actacacgaa tgtccagtcc cagctccagc caaacggagc tcactttgac cagtgccaaa
      841 attgcatctt ctggttacta ctttgtgcgt gagttacttg aaatcatctg ctttgttttg
      901 ttctatttca gcgagctctt gctaaaactg gagcagagtc tatcagtttg cttgagctgt
      961 gtcgaaacac aaaccgaaag caggcagcag caaagttcta cagctttttg gttcttaaga
     1021 agcagcaagc catcgagctc acacaggaag agccgtacag tgacatcatt gcaacccctg
     1081 gaccacggtt ccatattatc gtcgacgatt acaaggatga cgatgacaag ggttccggcg
     1141 gcgtgcaggt ggagactatc tccccaggag acgggcgcac cttccccaag cgcggccaga
     1201 cctgcgtggt gcactacacc gggatgcttg aagatggaaa gaaagttgat tcctcccggg
     1261 acagaaacaa gccctttaag tttatgctag gcaagcagga ggtgatccga ggctgggaag
     1321 aaggggttgc ccagatgagt gtgggtcaga gagccaaact gactatatct ccagattatg
     1381 cctatggtgc cactgggcac ccaggcatca tcccaccaca tgccactctc gtcttcgatg
     1441 tggagcttct aaaactggaa gtcgattatc cgtacgacgt accagactac gcagtgagca
     1501 agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa
     1561 acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga
     1621 ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca
     1681 ccttcggcta cggcctgcag tgcttcgccc gctaccccga ccacatgaag cagcacgact
     1741 tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg
     1801 acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca
     1861 tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt
     1921 acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg
     1981 tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc
     2041 agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagct
     2101 accagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt
     2161 tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaaggga tccggcgcaa
     2221 caaacttctc tctgctgaaa caagccggag atgtcgaaga gaatcctgga ccgatggcca
     2281 agcctttgtc tcaagaagaa tccaccctca ttgaaagagc aacggctaca atcaacagca
     2341 tccccatctc tgaagactac agcgtcgcca gcgcagctct ctctagcgac ggccgcatct
     2401 tcactggtgt caatgtatat cattttactg ggggaccttg tgcagaactc gtggtgctgg
     2461 gcactgctgc tgctgcggca gctggcaacc tgacttgtat cgtcgcgatc ggaaatgaga
     2521 acaggggcat cttgagcccc tgcggacggt gccgacaggt gcttctcgat ctgcatcctg
     2581 ggatcaaagc catagtgaag gacagtgatg gacagccgac ggcagttggg attcgtgaat
     2641 tgctgccctc tggttatgtg tgggagggct aaggtaccag ctagatgtgt tcgagctagt
     2701 gataactcac tagtacatac aaattgcccc cgtgtgcagg gcaccaaaac cctttaagaa
     2761 agtttttaga tttctgtttg tacaaaaatc tttgcctttt ctttcttctt tttcccccca
     2821 gtgtttctaa ttttgtcaac catattttta agggaaactg cttatttggg ttgggtttgt
     2881 attcctggag aaaacagtag cccaagaacc cagaagactt ttaacagttc agaacagatg
     2941 tgtgcaatat tggtgcatgt aagaatatgg agtaacagtc aaaaggcacc atttttaatg
     3001 ttagttttcc attactatgt tgaaaggaaa acctgcctag gaaaatgcct gacactttaa
     3061 gaactgtggt ttgagtccct tgacaggaag agaaaaatgt cttcccatca gtgaaaccaa
     3121 cggtctggtt aaccactgta gtagggatag tgtgtgaagc atcgtgcttg tcttcttgat
     3181 catgtttaaa gtaaatccac actgagcaag aatttgtgag gttgtgctga ctgatcttct
     3241 tcggctatta tgttctgaaa tcattccgtg agtcttagga cctctgatgc tatgcaccgc
     3301 agctcagaag gtaccgggcc ccccctcgag gtcgatcgac ggtatcgata agcttgatct
     3361 agagcggccg ccaccgcggt ggagctccag cttttgttcc ctttagtgag ggttaatttc
     3421 gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     3481 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     3541 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     3601 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     3661 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     3721 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     3781 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     3841 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     3901 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     3961 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     4021 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     4081 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     4141 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     4201 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     4261 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     4321 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     4381 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     4441 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     4501 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     4561 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     4621 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     4681 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     4741 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     4801 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     4861 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     4921 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     4981 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     5041 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     5101 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     5161 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     5221 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     5281 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     5341 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     5401 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     5461 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     5521 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     5581 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg
     5641 tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat
     5701 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg
     5761 tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga
     5821 gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag
     5881 aaaataccgc atcaggc
//
