LOCUS       sequence-201099-        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    115..136
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(151..156,157..174,175..181)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    189..205
                     /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     194..216
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     213..229
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     213..229
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1175..1495
                     /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             1502..1528
                     /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             1502..1528
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1532..1558
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1568..1624
                     /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             1625..2332
                     /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(1767..1785)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     complement(3260..3277)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3260..3276)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3269..3291)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(3485..3504)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3604..3626
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3664..3682)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3750..3854
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3855..3923,3924..4715)
                     /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(4073..4092)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4886..5474
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5375..5394
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5628..5645
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg
       61 cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt
      121 gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt
      181 gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc
      241 caagcttgca tgcctgcagg tcgacaagcc ctttcaggta gagccagtac ctgttcccca
      301 cactgaaagc tagggtgctg ggcagggtgg tgatatcaag taccatgggg cgcctgtggg
      361 ggcatcttat tcccactccc acttccatag ctctctgaag gtctgcttgg gttcctggtt
      421 acttactgca tgtgactgca ggtgttacat actaggggat gggagtatct gtcgggagaa
      481 acagggtaat gtgagtgaga gaaccccgca ctaaattcct agcatgttta cagagactca
      541 gagattgtat gtttggtagt gagtagtgtg ttgaattact gagagagtgc tcataagcac
      601 tgttggctac atctccaggg gacacttggt gctgcttata gaaaaatcaa gtgagtctgt
      661 gacagttggc atcactaaat atcttaaatc atctgtattt attctacttt ctggagtgtc
      721 tctgaaaata ctcaggcctt aaatttgcac ttagtacaac ccttctgttt ttgtaagagc
      781 taaccatgtg ttcatagtgt cagagatccc tgtgtgtcag ttctaaatca cgctcagaag
      841 ggtctccaga ggaggaagga gacacactca ctctagtaca gtctagtcta gtacagtggt
      901 tttccttcca agcagtgagc cattgaccgc ccctgattct ggtcccgttt gccaagtggg
      961 ttgatgtctg gtttttcctt gacagtgcac attcgaaggc tgcgggaagc gcttttcact
     1021 ggacttcaat ttgcgcacac atgtgcgaat ccataccgga gacaggccct atgtgtgccc
     1081 cttcgacggt tgtaataaga agtttgctca gtcaactaac ctgaaatctc acatcttaac
     1141 acacgctaaa gccaaaaaca accagggatc cggaggagtg caggtggaaa ccatctcccc
     1201 aggagacggg cgcaccttcc ccaagcgcgg ccagacctgc gtggtgcact acaccgggat
     1261 gcttgaagat ggaaagaaag ttgattcctc ccgggacaga aacaagccct ttaagtttat
     1321 gctaggcaag caggaggtga tccgaggctg ggaagaaggg gttgcccaga tgagtgtggg
     1381 tcagagagcc aaactgacta tatctccaga ttatgcctat ggtgccactg ggcacccagg
     1441 catcatccca ccacatgcca ctctcgtctt cgatgtggag cttctaaaac tggaaggcgg
     1501 ctacccctac gacgtgcccg actacgccgg ctatccgtat gatgtcccgg actatgcagg
     1561 ctccggagca acaaacttct ctctgctgaa acaagccgga gatgtcgaag agaatcctgg
     1621 accggtgagc aagggcgagg aggataacat ggccatcatc aaggagttca tgcgcttcaa
     1681 ggtgcacatg gagggctccg tgaacggcca cgagttcgag atcgagggcg agggcgaggg
     1741 ccgcccctac gagggcaccc agaccgccaa gctgaaggtg accaagggtg gccccctgcc
     1801 cttcgcctgg gacatcctgt cccctcagtt catgtacggc tccaaggcct acgtgaagca
     1861 ccccgccgac atccccgact acttgaagct gtccttcccc gagggcttca agtgggagcg
     1921 cgtgatgaac ttcgaggacg gcggcgtggt gaccgtgacc caggactcct ccctgcagga
     1981 cggcgagttc atctacaagg tgaagctgcg cggcaccaac ttcccctccg acggccccgt
     2041 aatgcagaag aaaaccatgg gctgggaggc ctcctccgag cggatgtacc ccgaggacgg
     2101 cgccctgaag ggcgagatca agcagaggct gaagctgaag gacggcggcc actacgacgc
     2161 tgaggtcaag accacctaca aggccaagaa gcccgtgcag ctgcccggcg cctacaacgt
     2221 caacatcaag ttggacatca cctcccacaa cgaggactac accatcgtgg aacagtacga
     2281 acgcgccgag ggccgccact ccaccggcgg catggacgag ctgtacaagt aaaaagaaga
     2341 gagaagacct tctcgacccg ggaagcctct tcaggagtgt gattgggaat aaatatgcct
     2401 ctcctttgta tattatttct aggaagaatt ttaaaaatga atcctacaca cttaagggac
     2461 atgttttgat aaagtagtaa aaatttaaaa aatactttaa taagatgaca ttgctaagat
     2521 gctatatctt gctctgtaat ctcgtttcaa aaacaaggtg tttttgtaaa gtgtggtccc
     2581 aacaggagga caattcatga acttcgcatc aaaagacaat tctttataca acagtgctaa
     2641 aaatgggact tcttttcaca ttcttataaa tatgaagctc acctgttgct tacaattttt
     2701 ttaattttgt attttccaag tgtgcatatt gtacactttt tggggatatg cttagtaatg
     2761 ctgtgtgatt ttctggaggt tgataacttt gcttgcggta gattttcttt aaaagaatgg
     2821 gcagttacat gcatacttca aaagtatttt tcctgtacaa aaaaaaagtt atataggttt
     2881 tgtttgctat cttaattttg gttgtattct ttgatgttaa cacattttgt ataattgtat
     2941 cgtatagctg tattgaatca tgtagaatca aatattagat gtgatttaat agtgttaatc
     3001 aatttaaacc cattttagtc actttttttt ccccaaaaaa tactgccaga tgctgatgtt
     3061 cagtgtaatt tctttgcctg ttcagttaca gaaagtggtg ctcagttgta gaatgtattg
     3121 taccttttaa catctgatgt gtacatccgt gtaacagaaa gggcaacaat aaaatagcga
     3181 tcctaaagaa agattacggc agaaagagct ctgtaagcac agccttattt tcggatcccc
     3241 gggtaccgag ctcgaattca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg
     3301 gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg
     3361 aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatggcgcc
     3421 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata tggtgcactc
     3481 tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg
     3541 ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg
     3601 tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa
     3661 agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga
     3721 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     3781 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     3841 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     3901 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     3961 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     4021 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     4081 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     4141 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     4201 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     4261 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     4321 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     4381 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     4441 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     4501 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     4561 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     4621 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     4681 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     4741 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     4801 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     4861 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     4921 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     4981 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     5041 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     5101 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     5161 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     5221 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     5281 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     5341 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     5401 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     5461 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     5521 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     5581 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     5641 gcgagga
//
