LOCUS       sequence-323549-        6508 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Acute Depletion of Mau2-EYFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6508)
  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 6508)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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-Mau2_BSD_FKBPF36V_EYFP
FEATURES             Location/Qualifiers
     source          1..6508
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     127..149
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             1463..1858
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     CDS             1868..1924
                     /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             1925..1948
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1958..2278
                     /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             2285..2311
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             join(2312..2314,2315..3025)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(2354..2375)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2615..2634)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2962..2983
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     promoter        complement(4011..4029)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4050..4066)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4050..4066)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4063..4085)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4074..4090
                     /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(4098..4104,4105..4122,4123..4128))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4143..4164
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4281..4298)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4452..5040)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4532..4551)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5211..6002,6003..6071))
                     /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     5834..5853
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6072..6176)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6244..6262
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6300..6322)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6422..6441
                     /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 ggtcgacctc tgaggcctat
      241 ccagcctctg tggtgaaacc tctgccttcc tatagacagc atctactcat atggaacatc
      301 ttgtactcaa gaaagacttt tcatggagcc agagagatgg ctcggaggtt aagagtgctt
      361 gtcactcttc cagagaatcc aagcaaacat ccaggtggcc tactataact ccagctccag
      421 gtactcacac acaaagacat aaagacaaat ggatttttaa aaactgttaa gccttccctg
      481 accttcaata tattagcagc atccaacctc acgtcacgag aactagttct ttgcaaaatt
      541 tgcttccagg tgcactgaca cgcaagctta gcacagcgtt tggacccagc aagtgtggaa
      601 gcatatggtg aatgggcaaa tgcatttcat cctaggggca cagggacagc taaacccgta
      661 accgtgggaa accgccctgg ctgacccagt gcagttcgga aacgcacatt tcccacaagg
      721 caggacccaa gccaaagggg accggagggg caggagggct gtcccaggct gtccgctgtg
      781 gatgccaccg gcccgagagc agtttattgg gaaagcccac gcggttccca cccggcgccc
      841 aggcctttga gggagaccag gagcgctcct cagccacggg cgggggttgg ggggcgaacc
      901 cgagtctggg gtctcggcag accagagaag ggcacacgca acgttactgt aagtttccct
      961 ggggctgagg cgagaagtcc gcgtgtcagg tccgtgacag aaagtagggt gaacaggaga
     1021 cgtgcagtcg cgagaaccca ggcgggcggc gaagggaagt cgggggatgt tacctgcaac
     1081 atcccggttg tccatcttga gactcatcca atcccacaat gctccggcgc cacctgagaa
     1141 gagggaggct ctgttatagt gcgcttggtc tgatggtaaa tgtagtcctt ctccgacaat
     1201 ctcgtccaac ccccttctag gacaccgggc gtcttctcca cagaaacgcc ggcgtcagtt
     1261 cacgaacgtg tattactctc agttacgatg atgcagaggt atcatcctct cctcagagtc
     1321 ttcctcagtc ggagccactt cgaacacaca ccataagacc cagaagtcct tgcgtagcag
     1381 ttctcgcgag aattgcgggt cgccaggccc cgcccctcaa ggtgcttccg cctctctgtg
     1441 gaggcggctt gttgttgtcg acatggccaa gcctttgtct caagaagaat ccaccctcat
     1501 tgaaagagca acggctacaa tcaacagcat ccccatctct gaagactaca gcgtcgccag
     1561 cgcagctctc tctagcgacg gccgcatctt cactggtgtc aatgtatatc attttactgg
     1621 gggaccttgt gcagaactcg tggtgctggg cactgctgct gctgcggcag ctggcaacct
     1681 gacttgtatc gtcgcgatcg gaaatgagaa caggggcatc ttgagcccct gcggacggtg
     1741 ccgacaggtg cttctcgatc tgcatcctgg gatcaaagcc atagtgaagg acagtgatgg
     1801 acagccgacg gcagttggga ttcgtgaatt gctgccctct ggttatgtgt gggagggcgg
     1861 ttccggagcc acgaacttct ctctgttaaa gcaagcagga gacgtggaag aaaaccccgg
     1921 tcccgattac aaggatgacg atgacaaggg ttccggcggc gtgcaggtgg agactatctc
     1981 cccaggagac gggcgcacct tccccaagcg cggccagacc tgcgtggtgc actacaccgg
     2041 gatgcttgaa gatggaaaga aagttgattc ctcccgggac agaaacaagc cctttaagtt
     2101 tatgctaggc aagcaggagg tgatccgagg ctgggaagaa ggggttgccc agatgagtgt
     2161 gggtcagaga gccaaactga ctatatctcc agattatgcc tatggtgcca ctgggcaccc
     2221 aggcatcatc ccaccacatg ccactctcgt cttcgatgtg gagcttctaa aactggaagt
     2281 cgattatccg tacgacgtac cagactacgc agtgagcaag ggcgaggagc tgttcaccgg
     2341 ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc
     2401 cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac
     2461 cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ttcggctacg gcctgcagtg
     2521 cttcgcccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga
     2581 aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc
     2641 cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt
     2701 caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt
     2761 ctatatcatg gccgacaagc agaagaacgg catcaaggtg aacttcaaga tccgccacaa
     2821 catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga
     2881 cggccccgtg ctgctgcccg acaaccacta cctgagctac cagtccgccc tgagcaaaga
     2941 ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac
     3001 tctcggcatg gacgagctgt acaagggtgg aggtggatct ggtggcggcg gttcaggtac
     3061 cgcggcacag gcggcggctg cggcgcaggc ggcggctgca gctcaggctg ccgccgcaca
     3121 ggccgcacag gccgaggcag ccgagtcgtg gtatctggct ctcctgggtt tcgccgagca
     3181 cttccgaaca tctagcccgc ccaagatccg cttgtgcgtt cattgcctgc aggccgtgtt
     3241 ccccttcaag ccgcctcagc ggatcgaggc ccggacgcac ctgcagctcg gctcggtgct
     3301 ttaccaccac acgaagaaca gcgagcaggc gcgcagccac ctggagaagg cggtgagtgt
     3361 gcgggccagg cgggcgggcc gcgggtaccg gggactcgat cctctccgcc tcgtcgggct
     3421 tgggacttcc gaaggcctcg ccctcacctg cggtgacaga cgcggctccg ccacgccctc
     3481 tcacccggga accgggaaag ttgtccctct tcagctactt ggtgcttgca tcggggaccc
     3541 tcctagtcgg gtggccgcgg gaacaggacc cagagcgtca gcctagaagc acctacctaa
     3601 ggaagagatg cgagctccct gacgtgtgaa agcgaggtcg caggaaatct gaaagtccag
     3661 gacttctgac agtcaggtgt ctaggtctgg agtcccagat ccttgtggca tggatattgg
     3721 ggaaggtgag gccttcagcc tcttgttgaa cgttgttgcc ttgtgaggac gtgaagtgca
     3781 gaaagtagct tctagagtca cttgcacctc gggggtctgg aggtcagtct cactgttggt
     3841 ggcttcttct cccaccacga aatggtccat ctttgaaaga gtccccactc cgcctccaac
     3901 ttgatttgtg acgtcttggt gcttcacagg gtaccctcga ggtcgatcga cggtatcgat
     3961 aagcttgatc tagagcggcc gccaccgcgg tggagctcca gcttttgttc cctttagtga
     4021 gggttaattt cgagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     4081 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     4141 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     4201 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     4261 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     4321 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     4381 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     4441 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     4501 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     4561 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     4621 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     4681 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     4741 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     4801 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     4861 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     4921 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     4981 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     5041 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     5101 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     5161 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     5221 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     5281 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     5341 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     5401 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     5461 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     5521 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     5581 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     5641 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     5701 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     5761 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     5821 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     5881 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     5941 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     6001 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     6061 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     6121 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     6181 tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat
     6241 aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac
     6301 ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc
     6361 agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat
     6421 gcggcatcag agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga
     6481 tgcgtaagga gaaaataccg catcaggc
//
