LOCUS       sequence-363948-        7234 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For the mammalian expression of the human protein ApoE3_D125I_47-300
            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 7234)
  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 7234)
  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_47-300_mCh-2xFKBP (pBS1114)
FEATURES             Location/Qualifiers
     source          1..7234
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(56..75)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        247..626
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        627..830
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     781..801
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    832..850
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    853..871
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     881..905
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      1014..1023
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2526..2846
                     /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             2853..3173
                     /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             3180..3206
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(3254..3271)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3260..3484
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3530..3958
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3617..3636)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3827..3848
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3967..3987)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3972..4301
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4152..4287
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4214..4233
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4349..4396
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4415..4789
                     /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    4919..5040
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4956..4975)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5010..5029
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5089..5105)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5089..5105)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5102..5124)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5113..5129
                     /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(5137..5143,5144..5161,5162..5167))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5182..5203
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5320..5337)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5491..6079)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5571..5590)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6250..7041,7042..7110))
                     /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     6873..6892
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7111..7215)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ccacctgacg tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa
       61 tctgctctga tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg
      121 ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca
      181 tgaagaatct gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata
      241 cgcgttgaca ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc
      301 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
      361 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
      421 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
      481 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc
      541 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct
      601 acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg
      661 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
      721 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga
      781 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct ctccctatca
      841 gtgatagaga tctccctatc agtgatagag atcgtcgacg agctcgttta gtgaaccgtc
      901 agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagacac cgggaccgat
      961 ccagcctccg gactctagcg tttaaactta agcttggtac cgagctcgga tccgccacca
     1021 tggccctggg gagattttgg gactatctga gatgggtgca gaccctgagc gagcaggtgc
     1081 aggaggagct cctgagctct caggtgacac aggaactgag agccctgatg gacgagacta
     1141 tgaaggaact gaaggcctac aaatctgagc tggaagagca gctgaccccc gtggctgaag
     1201 agacccgcgc tcgcttgtcc aaagagctcc aggccgccca ggcccggctg ggtgccatta
     1261 tggaggacgt ctgcggccgc ctggtccagt accgcgggga agtgcaggct atgctggggc
     1321 agtctacaga ggagctccgg gtcagactgg cctctcacct gcggaagctg aggaagaggt
     1381 tgctccgcga cgccgacgac ctccagaagc gcctggctgt gtaccaggcc ggcgcccggg
     1441 aaggcgccga gcgcggactc tccgctatca gggagcggct cggccctctg gttgaacagg
     1501 gcagagtcag ggctgccact gtggggagcc tcgccggaca gcccctccag gagagagccc
     1561 aggcctgggg cgagcgcctg cgagctagaa tggaggagat ggggtccaga acccgcgacc
     1621 gcctggacga agtgaaggag caggtggccg aggtgcgtgc caaactggag gaacaggccc
     1681 agcagatccg cctccaggct gaggccttcc aggccagact gaagagctgg tttgagcctc
     1741 ttgtggagga tatgcagcgg cagtgggccg ggctggtcga gaagtccgga ggcggcggat
     1801 ccggcggaat ggtgtctaaa ggcgaggagg ataacatggc tataatcaag gaatttatga
     1861 ggttcaaggt gcatatggag ggatctgtga acggtcacga gttcgaaatc gagggcgaag
     1921 gcgaggggcg accctatgaa gggacacaga cggccaaact gaaggtgacc aagggtggcc
     1981 ccctgccttt cgcctgggac atcctgtcac cccagtttat gtacggctct aaggcttacg
     2041 ttaagcaccc tgccgatatt ccggactact tgaaactgag cttccctgaa gggttcaagt
     2101 gggaacgggt aatgaatttt gaggatggcg gtgtagtcac agttacccag gacagctctc
     2161 ttcaggacgg agaatttatc tataaggtta aacttcgggg cactaacttc ccatcagacg
     2221 gccccgtgat gcagaagaaa actatgggct gggaagccag ctcagaacgc atgtatcccg
     2281 aggacggagc actgaaggga gaaatcaagc agcgactcaa gctgaaggat gggggacatt
     2341 atgatgctga ggtcaaaacc acctataagg ccaagaaacc cgttcagctt cctggggcat
     2401 acaacgtgaa tatcaaactg gatattacat cccataacga agactatacc atcgtggaac
     2461 agtacgagcg ggccgagggc agacatagca cagggggcat ggatgaattg tacaaggcga
     2521 ctagaggcgt ccaagtcgaa accattagtc ccggcgatgg cagaacattt cctaaaaggg
     2581 gacaaacatg tgtcgtccat tatacaggca tgttggagga cggcaaaaag gtggacagta
     2641 gtagagatcg caataaacct ttcaaattca tgttgggaaa acaagaagtc attaggggat
     2701 gggaggaggg cgtggctcaa atgtccgtcg gccaacgcgc taagctcacc atcagccccg
     2761 actacgcata cggcgctacc ggacatcccg gaattattcc ccctcacgct accttggtgt
     2821 ttgacgtcga actgttgaag ctcgagacta gaggagtgca ggtggagact atctccccag
     2881 gagacgggcg caccttcccc aagcgcggcc agacctgcgt ggtgcactac accgggatgc
     2941 ttgaagatgg aaagaaagtt gattcctccc gggacagaaa caagcccttt aagtttatgc
     3001 taggcaagca ggaggtgatc cgaggctggg aagaaggggt tgcccagatg agtgtgggtc
     3061 agagagccaa actgactata tctccagatt atgcctatgg tgccactggg cacccaggca
     3121 tcatcccacc acatgccact ctcgtcttcg atgtggagct tctaaaactg gaaactagtt
     3181 atccgtacga cgtaccagac tacgcataag cggccgctcg agtctagagg gcccgtttaa
     3241 acccgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc
     3301 cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag
     3361 gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag
     3421 gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct
     3481 atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt
     3541 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     3601 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     3661 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     3721 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     3781 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     3841 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     3901 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa
     3961 ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
     4021 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
     4081 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
     4141 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     4201 gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga
     4261 agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta
     4321 tatccatttt cggatctgat cagcacgtgt tgacaattaa tcatcggcat agtatatcgg
     4381 catagtataa tacgacaagg tgaggaacta aaccatggcc aagttgacca gtgccgttcc
     4441 ggtgctcacc gcgcgcgacg tcgccggagc ggtcgagttc tggaccgacc ggctcgggtt
     4501 ctcccgggac ttcgtggagg acgacttcgc cggtgtggtc cgggacgacg tgaccctgtt
     4561 catcagcgcg gtccaggacc aggtggtgcc ggacaacacc ctggcctggg tgtgggtgcg
     4621 cggcctggac gagctgtacg ccgagtggtc ggaggtcgtg tccacgaact tccgggacgc
     4681 ctccgggccg gccatgaccg agatcggcga gcagccgtgg gggcgggagt tcgccctgcg
     4741 cgacccggcc ggcaactgcg tgcacttcgt ggccgaggag caggactgac acgtgctacg
     4801 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
     4861 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa
     4921 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     4981 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     5041 tcatgtctgt ataccgtcga cctctagcta tagcttggcg taatcatggt catagctgtt
     5101 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     5161 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     5221 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     5281 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     5341 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     5401 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     5461 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     5521 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     5581 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     5641 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     5701 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     5761 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     5821 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     5881 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     5941 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     6001 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     6061 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     6121 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     6181 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     6241 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     6301 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     6361 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     6421 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     6481 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     6541 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     6601 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     6661 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     6721 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     6781 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     6841 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     6901 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     6961 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     7021 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     7081 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     7141 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     7201 aataggggtt ccgcgcacat ttccccgaaa agtg
//
