LOCUS       sequence-197858-        5647 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For knocking in FKBP F36V P2A mCherry onto YY1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5647)
  AUTHORS   Weintraub AS, Li CH, Zamudio AV, Sigova AA, Hannett NM, Day DS, 
            Abraham BJ, Cohen MA, Nabet B, Buckley DL, Guo YE, Hnisz D, Jaenisch
            R, Bradner JE, Gray NS, Young RA
  TITLE     YY1 Is a Structural Regulator of Enhancer-Promoter Loops.
  JOURNAL   Cell. 2017 Dec 14;171(7):1573-1588.e28. doi: 
            10.1016/j.cell.2017.11.008. Epub 2017 Dec 7.
  PUBMED    29224777
REFERENCE   2  (bases 1 to 5647)
  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"; doi: 
            "10.1016/j.cell.2017.11.008"; journalName: "Cell"; date: 
            "2017-12-14- 14"; volume: "171"; issue: "7"; pages: "1573-1588.e28"
COMMENT     Sequence Label: pAW62.YY1.FKBP.knock-in.mCherry
FEATURES             Location/Qualifiers
     source          1..5647
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    106..127
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(142..147,148..165,166..172)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    180..196
                     /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)."
     primer_bind     185..207
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     204..220
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     204..220
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1166..1486
                     /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             1493..1519
                     /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             1493..1519
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1523..1549
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1559..1615
                     /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             1616..2323
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     complement(1758..1776)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     complement(3251..3268)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3251..3267)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3260..3282)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(3476..3495)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3595..3617
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3655..3673)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3741..3845
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3846..3914,3915..4706)
                     /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     complement(4064..4083)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4877..5465
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5366..5385
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5619..5636
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
       61 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct
      121 cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat
      181 tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg ccaagcttgc
      241 atgcctgcag gtcgacaagc cctttcaggt agagccagta cctgttcccc acactgaaag
      301 ctagggtgct gggcagggtg gtgatatcaa gtaccatggg gcgcctgtgg gggcatctta
      361 ttcccactcc cacttccata gctctctgaa ggtctgcttg ggttcctggt tacttactgc
      421 atgtgactgc aggtgttaca tactagggga tgggagtatc tgtcgggaga aacagggtaa
      481 tgtgagtgag agaaccccgc actaaattcc tagcatgttt acagagactc agagattgta
      541 tgtttggtag tgagtagtgt gttgaattac tgagagagtg ctcataagca ctgttggcta
      601 catctccagg ggacacttgg tgctgcttat agaaaaatca agtgagtctg tgacagttgg
      661 catcactaaa tatcttaaat catctgtatt tattctactt tctggagtgt ctctgaaaat
      721 actcaggcct taaatttgca cttagtacaa cccttctgtt tttgtaagag ctaaccatgt
      781 gttcatagtg tcagagatcc ctgtgtgtca gttctaaatc acgctcagaa gggtctccag
      841 aggaggaagg agacacactc actctagtac agtctagtct agtacagtgg ttttccttcc
      901 aagcagtgag ccattgaccg cccctgattc tggtcccgtt tgccaagtgg gttgatgtct
      961 ggtttttcct tgacagtgca cattcgaagg ctgcgggaag cgcttttcac tggacttcaa
     1021 tttgcgcaca catgtgcgaa tccataccgg agacaggccc tatgtgtgcc ccttcgacgg
     1081 ttgtaataag aagtttgctc agtcaactaa cctgaaatct cacatcttaa cacacgctaa
     1141 agccaaaaac aaccagggat ccggaggagt gcaggtggaa accatctccc caggagacgg
     1201 gcgcaccttc cccaagcgcg gccagacctg cgtggtgcac tacaccggga tgcttgaaga
     1261 tggaaagaaa gttgattcct cccgggacag aaacaagccc tttaagttta tgctaggcaa
     1321 gcaggaggtg atccgaggct gggaagaagg ggttgcccag atgagtgtgg gtcagagagc
     1381 caaactgact atatctccag attatgccta tggtgccact gggcacccag gcatcatccc
     1441 accacatgcc actctcgtct tcgatgtgga gcttctaaaa ctggaaggcg gctaccccta
     1501 cgacgtgccc gactacgccg gctatccgta tgatgtcccg gactatgcag gctccggagc
     1561 aacaaacttc tctctgctga aacaagccgg agatgtcgaa gagaatcctg gaccggtgag
     1621 caagggcgag gaggataaca tggccatcat caaggagttc atgcgcttca aggtgcacat
     1681 ggagggctcc gtgaacggcc acgagttcga gatcgagggc gagggcgagg gccgccccta
     1741 cgagggcacc cagaccgcca agctgaaggt gaccaagggt ggccccctgc ccttcgcctg
     1801 ggacatcctg tcccctcagt tcatgtacgg ctccaaggcc tacgtgaagc accccgccga
     1861 catccccgac tacttgaagc tgtccttccc cgagggcttc aagtgggagc gcgtgatgaa
     1921 cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc tccctgcagg acggcgagtt
     1981 catctacaag gtgaagctgc gcggcaccaa cttcccctcc gacggccccg taatgcagaa
     2041 gaaaaccatg ggctgggagg cctcctccga gcggatgtac cccgaggacg gcgccctgaa
     2101 gggcgagatc aagcagaggc tgaagctgaa ggacggcggc cactacgacg ctgaggtcaa
     2161 gaccacctac aaggccaaga agcccgtgca gctgcccggc gcctacaacg tcaacatcaa
     2221 gttggacatc acctcccaca acgaggacta caccatcgtg gaacagtacg aacgcgccga
     2281 gggccgccac tccaccggcg gcatggacga gctgtacaag taaaaagaag agagaagacc
     2341 ttctcgaccc gggaagcctc ttcaggagtg tgattgggaa taaatatgcc tctcctttgt
     2401 atattatttc taggaagaat tttaaaaatg aatcctacac acttaaggga catgttttga
     2461 taaagtagta aaaatttaaa aaatacttta ataagatgac attgctaaga tgctatatct
     2521 tgctctgtaa tctcgtttca aaaacaaggt gtttttgtaa agtgtggtcc caacaggagg
     2581 acaattcatg aacttcgcat caaaagacaa ttctttatac aacagtgcta aaaatgggac
     2641 ttcttttcac attcttataa atatgaagct cacctgttgc ttacaatttt tttaattttg
     2701 tattttccaa gtgtgcatat tgtacacttt ttggggatat gcttagtaat gctgtgtgat
     2761 tttctggagg ttgataactt tgcttgcggt agattttctt taaaagaatg ggcagttaca
     2821 tgcatacttc aaaagtattt ttcctgtaca aaaaaaaagt tatataggtt ttgtttgcta
     2881 tcttaatttt ggttgtattc tttgatgtta acacattttg tataattgta tcgtatagct
     2941 gtattgaatc atgtagaatc aaatattaga tgtgatttaa tagtgttaat caatttaaac
     3001 ccattttagt cacttttttt tccccaaaaa atactgccag atgctgatgt tcagtgtaat
     3061 ttctttgcct gttcagttac agaaagtggt gctcagttgt agaatgtatt gtacctttta
     3121 acatctgatg tgtacatccg tgtaacagaa agggcaacaa taaaatagcg atcctaaaga
     3181 aagattacgg cagaaagagc tctgtaagca cagccttatt ttcggatccc cgggtaccga
     3241 gctcgaattc actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc
     3301 aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc
     3361 gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggcgc ctgatgcggt
     3421 attttctcct tacgcatctg tgcggtattt cacaccgcat atggtgcact ctcagtacaa
     3481 tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc
     3541 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga
     3601 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg
     3661 tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg
     3721 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     3781 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     3841 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     3901 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     3961 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     4021 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     4081 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     4141 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     4201 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     4261 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     4321 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     4381 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     4441 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     4501 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     4561 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     4621 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     4681 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     4741 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     4801 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     4861 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     4921 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     4981 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     5041 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     5101 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     5161 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     5221 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     5281 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     5341 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     5401 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     5461 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     5521 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     5581 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     5641 cggaaga
//
