LOCUS       sequence-201101-        5650 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For knocking in FKBP F36V P2A BFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5650)
  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 5650)
  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: pAW63.YY1.FKBP.knock-in.BFP
FEATURES             Location/Qualifiers
     source          1..5650
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(143..148,149..166,167..173)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /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     186..208
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     CDS             1167..1487
                     /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             1494..1520
                     /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             1494..1520
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1524..1550
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1560..1616
                     /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             1617..2324
                     /codon_start=1
                     /product="enhanced monomeric blue fluorescent protein 
                     (Subach et al., 2011)"
                     /label=mTagBFP2
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMR
                     IKVVEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGV
                     LTATQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDM
                     ALKLVGGSHLIANAKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVA
                     RYCDLPSKLGHKLN"
     primer_bind     complement(3255..3272)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3255..3271)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3264..3286)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(3480..3499)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3599..3621
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3659..3677)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3745..3849
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3850..3918,3919..4710)
                     /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(4068..4087)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4881..5469
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5370..5389
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5623..5640
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagatatga ccatgattac gccaagcttg
      241 catgcctgca ggtcgacaag ccctttcagg tagagccagt acctgttccc cacactgaaa
      301 gctagggtgc tgggcagggt ggtgatatca agtaccatgg ggcgcctgtg ggggcatctt
      361 attcccactc ccacttccat agctctctga aggtctgctt gggttcctgg ttacttactg
      421 catgtgactg caggtgttac atactagggg atgggagtat ctgtcgggag aaacagggta
      481 atgtgagtga gagaaccccg cactaaattc ctagcatgtt tacagagact cagagattgt
      541 atgtttggta gtgagtagtg tgttgaatta ctgagagagt gctcataagc actgttggct
      601 acatctccag gggacacttg gtgctgctta tagaaaaatc aagtgagtct gtgacagttg
      661 gcatcactaa atatcttaaa tcatctgtat ttattctact ttctggagtg tctctgaaaa
      721 tactcaggcc ttaaatttgc acttagtaca acccttctgt ttttgtaaga gctaaccatg
      781 tgttcatagt gtcagagatc cctgtgtgtc agttctaaat cacgctcaga agggtctcca
      841 gaggaggaag gagacacact cactctagta cagtctagtc tagtacagtg gttttccttc
      901 caagcagtga gccattgacc gcccctgatt ctggtcccgt ttgccaagtg ggttgatgtc
      961 tggtttttcc ttgacagtgc acattcgaag gctgcgggaa gcgcttttca ctggacttca
     1021 atttgcgcac acatgtgcga atccataccg gagacaggcc ctatgtgtgc cccttcgacg
     1081 gttgtaataa gaagtttgct cagtcaacta acctgaaatc tcacatctta acacacgcta
     1141 aagccaaaaa caaccaggga tccggaggag tgcaggtgga aaccatctcc ccaggagacg
     1201 ggcgcacctt ccccaagcgc ggccagacct gcgtggtgca ctacaccggg atgcttgaag
     1261 atggaaagaa agttgattcc tcccgggaca gaaacaagcc ctttaagttt atgctaggca
     1321 agcaggaggt gatccgaggc tgggaagaag gggttgccca gatgagtgtg ggtcagagag
     1381 ccaaactgac tatatctcca gattatgcct atggtgccac tgggcaccca ggcatcatcc
     1441 caccacatgc cactctcgtc ttcgatgtgg agcttctaaa actggaaggc ggctacccct
     1501 acgacgtgcc cgactacgcc ggctatccgt atgatgtccc ggactatgca ggctccggag
     1561 caacaaactt ctctctgctg aaacaagccg gagatgtcga agagaatcct ggaccggtgt
     1621 ctaagggcga agagctgatt aaggagaaca tgcacatgaa gctgtacatg gagggcaccg
     1681 tggacaacca tcacttcaag tgcacatccg agggcgaagg caagccctac gagggcaccc
     1741 agaccatgag aatcaaggtg gtcgagggcg gccctctccc cttcgccttc gacatcctgg
     1801 ctactagctt cctctacggc agcaagacct tcatcaacca cacccagggc atccccgact
     1861 tcttcaagca gtccttccct gagggcttca catgggagag agtcaccaca tacgaagacg
     1921 ggggcgtgct gaccgctacc caggacacca gcctccagga cggctgcctc atctacaacg
     1981 tcaagatcag aggggtgaac ttcacatcca acggccctgt gatgcagaag aaaacactcg
     2041 gctgggaggc cttcaccgag acgctgtacc ccgctgacgg cggcctggaa ggcagaaacg
     2101 acatggccct gaagctcgtg ggcgggagcc atctgatcgc aaacgccaag accacatata
     2161 gatccaagaa acccgctaag aacctcaaga tgcctggcgt ctactatgtg gactacagac
     2221 tggaaagaat caaggaggcc aacaacgaga cctacgtcga gcagcacgag gtggcagtgg
     2281 ccagatactg cgacctccct agcaaactgg ggcacaagct taattaaaaa gaagagagaa
     2341 gaccttctcg acccgggaag cctcttcagg agtgtgattg ggaataaata tgcctctcct
     2401 ttgtatatta tttctaggaa gaattttaaa aatgaatcct acacacttaa gggacatgtt
     2461 ttgataaagt agtaaaaatt taaaaaatac tttaataaga tgacattgct aagatgctat
     2521 atcttgctct gtaatctcgt ttcaaaaaca aggtgttttt gtaaagtgtg gtcccaacag
     2581 gaggacaatt catgaacttc gcatcaaaag acaattcttt atacaacagt gctaaaaatg
     2641 ggacttcttt tcacattctt ataaatatga agctcacctg ttgcttacaa tttttttaat
     2701 tttgtatttt ccaagtgtgc atattgtaca ctttttgggg atatgcttag taatgctgtg
     2761 tgattttctg gaggttgata actttgcttg cggtagattt tctttaaaag aatgggcagt
     2821 tacatgcata cttcaaaagt atttttcctg tacaaaaaaa aagttatata ggttttgttt
     2881 gctatcttaa ttttggttgt attctttgat gttaacacat tttgtataat tgtatcgtat
     2941 agctgtattg aatcatgtag aatcaaatat tagatgtgat ttaatagtgt taatcaattt
     3001 aaacccattt tagtcacttt tttttcccca aaaaatactg ccagatgctg atgttcagtg
     3061 taatttcttt gcctgttcag ttacagaaag tggtgctcag ttgtagaatg tattgtacct
     3121 tttaacatct gatgtgtaca tccgtgtaac agaaagggca acaataaaat agcgatccta
     3181 aagaaagatt acggcagaaa gagctctgta agcacagcct tattttcgga tccccgggta
     3241 ccgagctcga attcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt
     3301 acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag
     3361 gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg gcgcctgatg
     3421 cggtattttc tccttacgca tctgtgcggt atttcacacc gcatatggtg cactctcagt
     3481 acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac
     3541 gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc
     3601 gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc
     3661 ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca
     3721 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     3781 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     3841 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     3901 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     3961 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     4021 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     4081 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     4141 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     4201 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     4261 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     4321 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     4381 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     4441 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     4501 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     4561 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     4621 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     4681 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     4741 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     4801 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     4861 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     4921 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     4981 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     5041 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     5101 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     5161 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     5221 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     5281 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg catggtcgga
     5341 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     5401 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     5461 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     5521 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     5581 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     5641 gaagcggaag
//
