LOCUS       sequence-364006-        7420 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For the mammalian expression of the human protein ApoE3_D125I 
            attached to 2xFKBP. The FKBP can be used for inducible dimerization 
            and condensate formation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7420)
  AUTHORS   Veling MT, Nguyen DT, Thadani NN, Oster ME, Rollins NJ, Brock KP, 
            Bethel NP, Lim S, Baker D, Way JC, Marks DS, Chang RL, Silver PA
  TITLE     Natural and Designed Proteins Inspired by Extremotolerant Organisms 
            Can Form Condensates and Attenuate Apoptosis in Human Cells.
  JOURNAL   ACS Synth Biol. 2022 Mar 18;11(3):1292-1302. doi: 
            10.1021/acssynbio.1c00572. Epub 2022 Feb 18.
  PUBMED    35176859
REFERENCE   2  (bases 1 to 7420)
  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: 
            "10.1021/acssynbio.1c00572"; journalName: "ACS Synth Biol"; date: 
            "2022-03-18- 18"; volume: "11"; issue: "3"; pages: "1292-1302"
COMMENT     Sequence Label: ApoE3_D125I_mCh-2xFKBP (pBS1157)
FEATURES             Location/Qualifiers
     source          1..7420
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(59..78)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        250..629
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        630..833
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     784..804
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    835..853
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    856..874
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     884..908
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             2715..3035
                     /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             3042..3362
                     /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             3369..3395
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(3443..3460)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3449..3673
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3719..4147
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3806..3825)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4016..4037
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4156..4176)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4161..4490
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4341..4476
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4403..4422
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4538..4585
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4604..4978
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    5108..5229
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5145..5164)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5199..5218
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5278..5294)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5278..5294)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5291..5313)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5302..5318
                     /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(5326..5332,5333..5350,5351..5356))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5371..5392
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5509..5526)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5680..6268)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5760..5779)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6439..7230,7231..7299))
                     /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     7062..7081
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7300..7404)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtgccacctg acgtcgacgg atcgggagat ctcccgatcc cctatggtgc actctcagta
       61 caatctgctc tgatgccgca tagttaagcc agtatctgct ccctgcttgt gtgttggagg
      121 tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg accgacaatt
      181 gcatgaagaa tctgcttagg gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat
      241 atacgcgttg acattgatta ttgactagtt attaatagta atcaattacg gggtcattag
      301 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
      361 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
      421 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
      481 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
      541 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
      601 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
      661 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
      721 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
      781 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctctcccta
      841 tcagtgatag agatctccct atcagtgata gagatcgtcg acgagctcgt ttagtgaacc
      901 gtcagatcgc ctggagacgc catccacgct gttttgacct ccatagaaga caccgggacc
      961 gatccagcct ccggactcta gcgtttaaac ttaagcttgg taccgagctc ggatccgcca
     1021 ccatgaaagt gttatgggca gcattgttgg tgacattttt agctgggtgc caggccaaag
     1081 tggaacaagc agtggaaaca gaacctgaac ctgaactgag acagcagacc gaatggcaaa
     1141 gtggccagag atgggaactg gccctgggga gattttggga ctatctgaga tgggtgcaga
     1201 ccctgagcga gcaggtgcag gaggagctcc tgagctctca ggtgacacag gaactgagag
     1261 ccctgatgga cgagactatg aaggaactga aggcctacaa atctgagctg gaagagcagc
     1321 tgacccccgt ggctgaagag acccgcgctc gcttgtccaa agagctccag gccgcccagg
     1381 cccggctggg tgccattatg gaggacgtct gcggccgcct ggtccagtac cgcggggaag
     1441 tgcaggctat gctggggcag tctacagagg agctccgggt cagactggcc tctcacctgc
     1501 ggaagctgag gaagaggttg ctccgcgacg ccgacgacct ccagaagcgc ctggctgtgt
     1561 accaggccgg cgcccgggaa ggcgccgagc gcggactctc cgctatcagg gagcggctcg
     1621 gccctctggt tgaacagggc agagtcaggg ctgccactgt ggggagcctc gccggacagc
     1681 ccctccagga gagagcccag gcctggggcg agcgcctgcg agctagaatg gaggagatgg
     1741 ggtccagaac ccgcgaccgc ctggacgaag tgaaggagca ggtggccgag gtgcgtgcca
     1801 aactggagga acaggcccag cagatccgcc tccaggctga ggccttccag gccagactga
     1861 agagctggtt tgagcctctt gtggaggata tgcagcggca gtgggccggg ctggtcgaga
     1921 aggtgcaggc cgccgtgggc acttccgccg cccctgtccc cagcgacaac cattccggag
     1981 gcggcggatc cggcggaatg gtgtctaaag gcgaggagga taacatggct ataatcaagg
     2041 aatttatgag gttcaaggtg catatggagg gatctgtgaa cggtcacgag ttcgaaatcg
     2101 agggcgaagg cgaggggcga ccctatgaag ggacacagac ggccaaactg aaggtgacca
     2161 agggtggccc cctgcctttc gcctgggaca tcctgtcacc ccagtttatg tacggctcta
     2221 aggcttacgt taagcaccct gccgatattc cggactactt gaaactgagc ttccctgaag
     2281 ggttcaagtg ggaacgggta atgaattttg aggatggcgg tgtagtcaca gttacccagg
     2341 acagctctct tcaggacgga gaatttatct ataaggttaa acttcggggc actaacttcc
     2401 catcagacgg ccccgtgatg cagaagaaaa ctatgggctg ggaagccagc tcagaacgca
     2461 tgtatcccga ggacggagca ctgaagggag aaatcaagca gcgactcaag ctgaaggatg
     2521 ggggacatta tgatgctgag gtcaaaacca cctataaggc caagaaaccc gttcagcttc
     2581 ctggggcata caacgtgaat atcaaactgg atattacatc ccataacgaa gactatacca
     2641 tcgtggaaca gtacgagcgg gccgagggca gacatagcac agggggcatg gatgaattgt
     2701 acaaggcgac tagaggcgtc caagtcgaaa ccattagtcc cggcgatggc agaacatttc
     2761 ctaaaagggg acaaacatgt gtcgtccatt atacaggcat gttggaggac ggcaaaaagg
     2821 tggacagtag tagagatcgc aataaacctt tcaaattcat gttgggaaaa caagaagtca
     2881 ttaggggatg ggaggagggc gtggctcaaa tgtccgtcgg ccaacgcgct aagctcacca
     2941 tcagccccga ctacgcatac ggcgctaccg gacatcccgg aattattccc cctcacgcta
     3001 ccttggtgtt tgacgtcgaa ctgttgaagc tcgagactag aggagtgcag gtggagacta
     3061 tctccccagg agacgggcgc accttcccca agcgcggcca gacctgcgtg gtgcactaca
     3121 ccgggatgct tgaagatgga aagaaagttg attcctcccg ggacagaaac aagcccttta
     3181 agtttatgct aggcaagcag gaggtgatcc gaggctggga agaaggggtt gcccagatga
     3241 gtgtgggtca gagagccaaa ctgactatat ctccagatta tgcctatggt gccactgggc
     3301 acccaggcat catcccacca catgccactc tcgtcttcga tgtggagctt ctaaaactgg
     3361 aaactagtta tccgtacgac gtaccagact acgcataagc ggccgctcga gtctagaggg
     3421 cccgtttaaa cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt
     3481 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
     3541 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
     3601 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg
     3661 gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac
     3721 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     3781 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     3841 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     3901 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     3961 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     4021 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     4081 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     4141 gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     4201 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     4261 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     4321 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     4381 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc
     4441 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg
     4501 gagcttgtat atccattttc ggatctgatc agcacgtgtt gacaattaat catcggcata
     4561 gtatatcggc atagtataat acgacaaggt gaggaactaa accatggcca agttgaccag
     4621 tgccgttccg gtgctcaccg cgcgcgacgt cgccggagcg gtcgagttct ggaccgaccg
     4681 gctcgggttc tcccgggact tcgtggagga cgacttcgcc ggtgtggtcc gggacgacgt
     4741 gaccctgttc atcagcgcgg tccaggacca ggtggtgccg gacaacaccc tggcctgggt
     4801 gtgggtgcgc ggcctggacg agctgtacgc cgagtggtcg gaggtcgtgt ccacgaactt
     4861 ccgggacgcc tccgggccgg ccatgaccga gatcggcgag cagccgtggg ggcgggagtt
     4921 cgccctgcgc gacccggccg gcaactgcgt gcacttcgtg gccgaggagc aggactgaca
     4981 cgtgctacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
     5041 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     5101 ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     5161 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     5221 tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc
     5281 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     5341 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     5401 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     5461 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     5521 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     5581 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     5641 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     5701 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     5761 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     5821 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     5881 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     5941 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     6001 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     6061 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     6121 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     6181 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     6241 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     6301 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     6361 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     6421 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     6481 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     6541 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     6601 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     6661 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     6721 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     6781 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     6841 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     6901 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     6961 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     7021 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     7081 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     7141 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     7201 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     7261 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     7321 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     7381 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
//
