LOCUS       sequence-323505-        6931 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Acute Depletion of Mau2-mScarlet.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6931)
  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 6931)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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_Neo_FKBPF36V_mScarlet
FEATURES             Location/Qualifiers
     source          1..6931
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     127..149
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             1463..2254
                     /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(1517..1536)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2127..2146
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             2264..2320
                     /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             2321..2347
                     /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             2321..2347
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2351..2377
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2387..2707
                     /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             2738..3433
                     /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(2871..2889)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3139..3158
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     promoter        complement(4434..4452)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4473..4489)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4473..4489)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4486..4508)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4497..4513
                     /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(4521..4527,4528..4545,4546..4551))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4566..4587
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4704..4721)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4875..5463)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4955..4974)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5634..6425,6426..6494))
                     /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     6257..6276
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6495..6599)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6667..6685
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6723..6745)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6845..6864
                     /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 acatgattga acaagatgga ttgcacgcag gttctccggc
     1501 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga
     1561 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
     1621 gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac
     1681 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
     1741 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt
     1801 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
     1861 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt
     1921 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag
     1981 gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt
     2041 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
     2101 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg
     2161 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg
     2221 catcgccttc tatcgccttc ttgacgagtt cttcggttcc ggagctacta acttcagcct
     2281 gctgaagcag gctggagacg tggaggagaa ccctggacct tacccctacg acgtgcccga
     2341 ctacgccggc tacccatacg atgttccaga ttacgctgga agcggaggag tgcaggtgga
     2401 aaccatctcc ccaggagacg ggcgcacctt ccccaagcgc ggccagacct gcgtggtgca
     2461 ctacaccggg atgcttgaag atggaaagaa agttgattcc tcccgggaca gaaacaagcc
     2521 ctttaagttt atgctaggca agcaggaggt gatccgaggc tgggaagaag gggttgccca
     2581 gatgagtgtg ggtcagagag ccaaactgac tatatctcca gattatgcct atggtgccac
     2641 tgggcaccca ggcatcatcc caccacatgc cactctcgtc ttcgatgtgg agcttctaaa
     2701 actggaaggt ggaggcggga gtggaggtgg aggatcgatg gtgagcaagg gcgaggcagt
     2761 gatcaaggag ttcatgcggt tcaaggtgca catggagggc tccatgaacg gccacgagtt
     2821 cgagatcgag ggcgagggcg agggccgccc ctacgagggc acccagaccg ccaagctgaa
     2881 ggtgaccaag ggtggccccc tgcccttctc ctgggacatc ctgtcccctc agttcatgta
     2941 cggctccagg gccttcatca agcaccccgc cgacatcccc gactactata agcagtcctt
     3001 ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag gacggcggcg ccgtgaccgt
     3061 gacccaggac acctccctgg aggacggcac cctgatctac aaggtgaagc tccgcggcac
     3121 caacttccct cctgacggcc ccgtaatgca gaagaagaca atgggctggg aagcgtccac
     3181 cgagcggttg taccccgagg acggcgtgct gaagggcgac attaagatgg ccctgcgcct
     3241 gaaggacggc ggccgctacc tggcggactt caagaccacc tacaaggcca agaagcccgt
     3301 gcagatgccc ggcgcctaca acgtggaccg caagttggac atcacctccc acaacgagga
     3361 ctacaccgtg gtggaacagt acgaacgctc cgagggccgc cactccaccg gcggcatgga
     3421 cgagctgtac aagggtggcg gtggctcggg cggtggtggg tcgggtggcg gcggatctgg
     3481 taccgcggca caggcggcgg ctgcggcgca ggcggcggct gcagctcagg ctgccgccgc
     3541 acaggccgca caggccgagg cagccgagtc gtggtatctg gctctcctgg gtttcgccga
     3601 gcacttccga acatctagcc cgcccaagat ccgcttgtgc gttcattgcc tgcaggccgt
     3661 gttccccttc aagccgcctc agcggatcga ggcccggacg cacctgcagc tcggctcggt
     3721 gctttaccac cacacgaaga acagcgagca ggcgcgcagc cacctggaga aggcggtgag
     3781 tgtgcgggcc aggcgggcgg gccgcgggta ccggggactc gatcctctcc gcctcgtcgg
     3841 gcttgggact tccgaaggcc tcgccctcac ctgcggtgac agacgcggct ccgccacgcc
     3901 ctctcacccg ggaaccggga aagttgtccc tcttcagcta cttggtgctt gcatcgggga
     3961 ccctcctagt cgggtggccg cgggaacagg acccagagcg tcagcctaga agcacctacc
     4021 taaggaagag atgcgagctc cctgacgtgt gaaagcgagg tcgcaggaaa tctgaaagtc
     4081 caggacttct gacagtcagg tgtctaggtc tggagtccca gatccttgtg gcatggatat
     4141 tggggaaggt gaggccttca gcctcttgtt gaacgttgtt gccttgtgag gacgtgaagt
     4201 gcagaaagta gcttctagag tcacttgcac ctcgggggtc tggaggtcag tctcactgtt
     4261 ggtggcttct tctcccacca cgaaatggtc catctttgaa agagtcccca ctccgcctcc
     4321 aacttgattt gtgacgtctt ggtgcttcac agggtaccct cgaggtcgat cgacggtatc
     4381 gataagcttg atctagagcg gccgccaccg cggtggagct ccagcttttg ttccctttag
     4441 tgagggttaa tttcgagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     4501 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     4561 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     4621 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     4681 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     4741 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     4801 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     4861 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     4921 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     4981 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     5041 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     5101 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     5161 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     5221 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     5281 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     5341 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     5401 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     5461 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     5521 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     5581 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     5641 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     5701 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     5761 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     5821 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     5881 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     5941 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     6001 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     6061 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     6121 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     6181 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     6241 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     6301 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     6361 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     6421 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     6481 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     6541 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     6601 acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc
     6661 tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa
     6721 aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg
     6781 agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac
     6841 tatgcggcat cagagcagat tgtactgaga gtgcaccata tgcggtgtga aataccgcac
     6901 agatgcgtaa ggagaaaata ccgcatcagg c
//
