LOCUS       sequence-363796-        5864 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For the mammalian expression of the human protein APOE_HUMAN_T243A. 
            This is one of the top performing targets in our apoptosis assay (it
            protected from apoptosis)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5864)
  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 5864)
  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: pST_APOE_HUMAN_T243A (pBS0818)
FEATURES             Location/Qualifiers
     source          1..5864
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(51..70)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        242..621
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        622..825
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     776..796
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     870..889
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        870..888
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1879..1896)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1885..2109
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2155..2583
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2242..2261)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2452..2473
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2592..2612)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2597..2926
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2777..2912
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2839..2858
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        2974..3021
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3040..3414
                     /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    3544..3665
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3581..3600)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3635..3654
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3714..3730)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3714..3730)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3727..3749)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3738..3754
                     /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(3762..3768,3769..3786,3787..3792))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(3945..3962)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4116..4704)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4196..4215)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4875..5666,5667..5735))
                     /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     5498..5517
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5736..5840)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tgacgtcgac ggatcgggag atctcccgat cccctatggt gcactctcag tacaatctgc
       61 tctgatgccg catagttaag ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag
      121 tagtgcgcga gcaaaattta agctacaaca aggcaaggct tgaccgacaa ttgcatgaag
      181 aatctgctta gggttaggcg ttttgcgctg cttcgcgatg tacgggccag atatacgcgt
      241 tgacattgat tattgactag ttattaatag taatcaatta cggggtcatt agttcatagc
      301 ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc
      361 aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg
      421 actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat
      481 caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc
      541 tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta catctacgta
      601 ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag
      661 cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt
      721 tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa
      781 atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctctctg gctaactaga
      841 gaacccactg cttactggct tatcgaaatt aatacgactc actataggga gacccaagct
      901 ggctagccac catgaaagtg ttatgggcag cattgttggt gacattttta gctgggtgcc
      961 aggccaaagt ggaacaagca gtggaaacag aacctgaacc tgaactgaga cagcagaccg
     1021 aatggcaaag tggccagaga tgggaactgg ccctggggag attttgggac tatctgagat
     1081 gggtgcagac cctgagcgag caggtgcagg aggagctcct gagctctcag gtgacacagg
     1141 aactgagagc cctgatggac gagactatga aggaactgaa ggcctacaaa tctgagctgg
     1201 aagagcagct gacccccgtg gctgaagaga cccgcgctcg cttgtccaaa gagctccagg
     1261 ccgcccaggc ccggctgggt gccgatatgg aggacgtctg cggccgcctg gtccagtacc
     1321 gcggggaagt gcaggctatg ctggggcagt ctacagagga gctccgggtc agactggcct
     1381 ctcacctgcg gaagctgagg aagaggttgc tccgcgacgc cgacgacctc cagaagcgcc
     1441 tggctgtgta ccaggccggc gcccgggaag gcgccgagcg cggactctcc gctatcaggg
     1501 agcggctcgg ccctctggtt gaacagggca gagtcagggc tgccactgtg gggagcctcg
     1561 ccggacagcc cctccaggag agagcccagg cctggggcga gcgcctgcga gctagaatgg
     1621 aggagatggg gtccagagcc cgcgaccgcc tggacgaagt gaaggagcag gtggccgagg
     1681 tgcgtgccaa actggaggaa caggcccagc agatccgcct ccaggctgag gccttccagg
     1741 ccagactgaa gagctggttt gagcctcttg tggaggatat gcagcggcag tgggccgggc
     1801 tggtcgagaa ggtgcaggcc gccgtgggca cttccgccgc ccctgtcccc agcgacaacc
     1861 attgaacccg ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc
     1921 cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa
     1981 atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg
     2041 ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg
     2101 gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc
     2161 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     2221 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     2281 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     2341 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
     2401 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     2461 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
     2521 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     2581 attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
     2641 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
     2701 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
     2761 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     2821 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt
     2881 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc
     2941 ttgtatatcc attttcggat ctgatcagca cgtgttgaca attaatcatc ggcatagtat
     3001 atcggcatag tataatacga caaggtgagg aactaaacca tggccaagtt gaccagtgcc
     3061 gttccggtgc tcaccgcgcg cgacgtcgcc ggagcggtcg agttctggac cgaccggctc
     3121 gggttctccc gggacttcgt ggaggacgac ttcgccggtg tggtccggga cgacgtgacc
     3181 ctgttcatca gcgcggtcca ggaccaggtg gtgccggaca acaccctggc ctgggtgtgg
     3241 gtgcgcggcc tggacgagct gtacgccgag tggtcggagg tcgtgtccac gaacttccgg
     3301 gacgcctccg ggccggccat gaccgagatc ggcgagcagc cgtgggggcg ggagttcgcc
     3361 ctgcgcgacc cggccggcaa ctgcgtgcac ttcgtggccg aggagcagga ctgacacgtg
     3421 ctacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc
     3481 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac
     3541 cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     3601 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     3661 tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag
     3721 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     3781 ataaagtgta aagcctgggg tgcctaatga gtgagctaac ttacattaat tgcgttgcgc
     3841 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     3901 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     3961 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     4021 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     4081 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     4141 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     4201 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     4261 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     4321 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     4381 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     4441 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     4501 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     4561 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     4621 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     4681 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     4741 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     4801 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     4861 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     4921 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     4981 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     5041 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     5101 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     5161 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     5221 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     5281 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     5341 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     5401 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     5461 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     5521 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     5581 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     5641 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     5701 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     5761 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     5821 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacc
//
