LOCUS       sequence-364004-        7420 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For the mammalian expression of the human protein ApoE3 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_mCh-2xFKBP (pBS1156)
FEATURES             Location/Qualifiers
     source          1..7420
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(69..88)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        260..639
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        640..843
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     794..814
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    845..863
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    866..884
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     894..918
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             2725..3045
                     /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             3052..3372
                     /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             3379..3405
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(3453..3470)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3459..3683
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3729..4157
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3816..3835)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4026..4047
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4166..4186)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4171..4500
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4351..4486
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4413..4432
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4548..4595
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4614..4988
                     /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    5118..5239
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5155..5174)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5209..5228
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5288..5304)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5288..5304)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5301..5323)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5312..5328
                     /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(5336..5342,5343..5360,5361..5366))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5381..5402
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5519..5536)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5690..6278)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5770..5789)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6449..7240,7241..7309))
                     /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     7072..7091
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7310..7414)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tccccgaaaa gtgccacctg acgtcgacgg atcgggagat ctcccgatcc cctatggtgc
       61 actctcagta caatctgctc tgatgccgca tagttaagcc agtatctgct ccctgcttgt
      121 gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg
      181 accgacaatt gcatgaagaa tctgcttagg gttaggcgtt ttgcgctgct tcgcgatgta
      241 cgggccagat atacgcgttg acattgatta ttgactagtt attaatagta atcaattacg
      301 gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc
      361 ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc
      421 atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact
      481 gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat
      541 gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact
      601 tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac
      661 atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac
      721 gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac
      781 tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga
      841 gctctcccta tcagtgatag agatctccct atcagtgata gagatcgtcg acgagctcgt
      901 ttagtgaacc gtcagatcgc ctggagacgc catccacgct gttttgacct ccatagaaga
      961 caccgggacc gatccagcct ccggactcta gcgtttaaac ttaagcttgg taccgagctc
     1021 ggatccgcca ccatgaaagt gttatgggca gcattgttgg tgacattttt agctgggtgc
     1081 caggccaaag tggaacaagc agtggaaaca gaacctgaac ctgaactgag acagcagacc
     1141 gaatggcaaa gtggccagag atgggaactg gccctgggga gattttggga ctatctgaga
     1201 tgggtgcaga ccctgagcga gcaggtgcag gaggagctcc tgagctctca ggtgacacag
     1261 gaactgagag ccctgatgga cgagactatg aaggaactga aggcctacaa atctgagctg
     1321 gaagagcagc tgacccccgt ggctgaagag acccgcgctc gcttgtccaa agagctccag
     1381 gccgcccagg cccggctggg tgccgatatg gaggacgtct gcggccgcct ggtccagtac
     1441 cgcggggaag tgcaggctat gctggggcag tctacagagg agctccgggt cagactggcc
     1501 tctcacctgc ggaagctgag gaagaggttg ctccgcgacg ccgacgacct ccagaagcgc
     1561 ctggctgtgt accaggccgg cgcccgggaa ggcgccgagc gcggactctc cgctatcagg
     1621 gagcggctcg gccctctggt tgaacagggc agagtcaggg ctgccactgt ggggagcctc
     1681 gccggacagc ccctccagga gagagcccag gcctggggcg agcgcctgcg agctagaatg
     1741 gaggagatgg ggtccagaac ccgcgaccgc ctggacgaag tgaaggagca ggtggccgag
     1801 gtgcgtgcca aactggagga acaggcccag cagatccgcc tccaggctga ggccttccag
     1861 gccagactga agagctggtt tgagcctctt gtggaggata tgcagcggca gtgggccggg
     1921 ctggtcgaga aggtgcaggc cgccgtgggc acttccgccg cccctgtccc cagcgacaac
     1981 cattccggag gcggcggatc cggcggaatg gtgtctaaag gcgaggagga taacatggct
     2041 ataatcaagg aatttatgag gttcaaggtg catatggagg gatctgtgaa cggtcacgag
     2101 ttcgaaatcg agggcgaagg cgaggggcga ccctatgaag ggacacagac ggccaaactg
     2161 aaggtgacca agggtggccc cctgcctttc gcctgggaca tcctgtcacc ccagtttatg
     2221 tacggctcta aggcttacgt taagcaccct gccgatattc cggactactt gaaactgagc
     2281 ttccctgaag ggttcaagtg ggaacgggta atgaattttg aggatggcgg tgtagtcaca
     2341 gttacccagg acagctctct tcaggacgga gaatttatct ataaggttaa acttcggggc
     2401 actaacttcc catcagacgg ccccgtgatg cagaagaaaa ctatgggctg ggaagccagc
     2461 tcagaacgca tgtatcccga ggacggagca ctgaagggag aaatcaagca gcgactcaag
     2521 ctgaaggatg ggggacatta tgatgctgag gtcaaaacca cctataaggc caagaaaccc
     2581 gttcagcttc ctggggcata caacgtgaat atcaaactgg atattacatc ccataacgaa
     2641 gactatacca tcgtggaaca gtacgagcgg gccgagggca gacatagcac agggggcatg
     2701 gatgaattgt acaaggcgac tagaggcgtc caagtcgaaa ccattagtcc cggcgatggc
     2761 agaacatttc ctaaaagggg acaaacatgt gtcgtccatt atacaggcat gttggaggac
     2821 ggcaaaaagg tggacagtag tagagatcgc aataaacctt tcaaattcat gttgggaaaa
     2881 caagaagtca ttaggggatg ggaggagggc gtggctcaaa tgtccgtcgg ccaacgcgct
     2941 aagctcacca tcagccccga ctacgcatac ggcgctaccg gacatcccgg aattattccc
     3001 cctcacgcta ccttggtgtt tgacgtcgaa ctgttgaagc tcgagactag aggagtgcag
     3061 gtggagacta tctccccagg agacgggcgc accttcccca agcgcggcca gacctgcgtg
     3121 gtgcactaca ccgggatgct tgaagatgga aagaaagttg attcctcccg ggacagaaac
     3181 aagcccttta agtttatgct aggcaagcag gaggtgatcc gaggctggga agaaggggtt
     3241 gcccagatga gtgtgggtca gagagccaaa ctgactatat ctccagatta tgcctatggt
     3301 gccactgggc acccaggcat catcccacca catgccactc tcgtcttcga tgtggagctt
     3361 ctaaaactgg aaactagtta tccgtacgac gtaccagact acgcataagc ggccgctcga
     3421 gtctagaggg cccgtttaaa cccgctgatc agcctcgact gtgccttcta gttgccagcc
     3481 atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt
     3541 cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct
     3601 ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc
     3661 tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg
     3721 gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     3781 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     3841 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     3901 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     3961 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     4021 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     4081 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
     4141 aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     4201 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     4261 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     4321 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     4381 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct
     4441 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     4501 aagctcccgg gagcttgtat atccattttc ggatctgatc agcacgtgtt gacaattaat
     4561 catcggcata gtatatcggc atagtataat acgacaaggt gaggaactaa accatggcca
     4621 agttgaccag tgccgttccg gtgctcaccg cgcgcgacgt cgccggagcg gtcgagttct
     4681 ggaccgaccg gctcgggttc tcccgggact tcgtggagga cgacttcgcc ggtgtggtcc
     4741 gggacgacgt gaccctgttc atcagcgcgg tccaggacca ggtggtgccg gacaacaccc
     4801 tggcctgggt gtgggtgcgc ggcctggacg agctgtacgc cgagtggtcg gaggtcgtgt
     4861 ccacgaactt ccgggacgcc tccgggccgg ccatgaccga gatcggcgag cagccgtggg
     4921 ggcgggagtt cgccctgcgc gacccggccg gcaactgcgt gcacttcgtg gccgaggagc
     4981 aggactgaca cgtgctacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct
     5041 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
     5101 agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     5161 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     5221 aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt
     5281 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     5341 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     5401 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     5461 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     5521 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     5581 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     5641 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     5701 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     5761 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     5821 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     5881 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     5941 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     6001 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     6061 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     6121 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     6181 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     6241 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     6301 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     6361 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     6421 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     6481 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     6541 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     6601 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     6661 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     6721 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     6781 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     6841 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     6901 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     6961 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     7021 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     7081 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     7141 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     7201 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     7261 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     7321 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     7381 gtatttagaa aaataaacaa ataggggttc cgcgcacatt
//
