LOCUS       sequence-363803-        5864 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For the mammalian expression of the human protein APOE_HUMAN_E114C. 
            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_E114C (pBS0838)
FEATURES             Location/Qualifiers
     source          1..5864
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(55..74)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        246..625
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        626..829
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     780..800
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     874..893
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        874..892
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1883..1900)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1889..2113
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2159..2587
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2246..2265)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2456..2477
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2596..2616)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2601..2930
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2781..2916
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2843..2862
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        2978..3025
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3044..3418
                     /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    3548..3669
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3585..3604)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3639..3658
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3718..3734)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3718..3734)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3731..3753)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3742..3758
                     /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(3766..3772,3773..3790,3791..3796))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(3949..3966)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4120..4708)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4200..4219)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4879..5670,5671..5739))
                     /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     5502..5521
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5740..5844)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cacctgacgt cgacggatcg ggagatctcc cgatccccta tggtgcactc tcagtacaat
       61 ctgctctgat gccgcatagt taagccagta tctgctccct gcttgtgtgt tggaggtcgc
      121 tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg acaattgcat
      181 gaagaatctg cttagggtta ggcgttttgc gctgcttcgc gatgtacggg ccagatatac
      241 gcgttgacat tgattattga ctagttatta atagtaatca attacggggt cattagttca
      301 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc
      361 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
      421 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
      481 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc
      541 cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta
      601 cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg
      661 atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt
      721 gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac
      781 gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctc tctggctaac
      841 tagagaaccc actgcttact ggcttatcga aattaatacg actcactata gggagaccca
      901 agctggctag ccaccatgaa agtgttatgg gcagcattgt tggtgacatt tttagctggg
      961 tgccaggcca aagtggaaca agcagtggaa acagaacctg aacctgaact gagacagcag
     1021 accgaatggc aaagtggcca gagatgggaa ctggccctgg ggagattttg ggactatctg
     1081 agatgggtgc agaccctgag cgagcaggtg caggaggagc tcctgagctc tcaggtgaca
     1141 caggaactga gagccctgat ggacgagact atgaaggaac tgaaggccta caaatctgag
     1201 ctggaagagc agctgacccc cgtggctgaa gagacccgcg ctcgcttgtc caaatgcctc
     1261 caggccgccc aggcccggct gggtgccgat atggaggacg tctgcggccg cctggtccag
     1321 taccgcgggg aagtgcaggc tatgctgggg cagtctacag aggagctccg ggtcagactg
     1381 gcctctcacc tgcggaagct gaggaagagg ttgctccgcg acgccgacga cctccagaag
     1441 cgcctggctg tgtaccaggc cggcgcccgg gaaggcgccg agcgcggact ctccgctatc
     1501 agggagcggc tcggccctct ggttgaacag ggcagagtca gggctgccac tgtggggagc
     1561 ctcgccggac agcccctcca ggagagagcc caggcctggg gcgagcgcct gcgagctaga
     1621 atggaggaga tggggtccag aacccgcgac cgcctggacg aagtgaagga gcaggtggcc
     1681 gaggtgcgtg ccaaactgga ggaacaggcc cagcagatcc gcctccaggc tgaggccttc
     1741 caggccagac tgaagagctg gtttgagcct cttgtggagg atatgcagcg gcagtgggcc
     1801 gggctggtcg agaaggtgca ggccgccgtg ggcacttccg ccgcccctgt ccccagcgac
     1861 aaccattgaa cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt
     1921 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
     1981 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
     2041 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg
     2101 gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac
     2161 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     2221 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     2281 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     2341 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     2401 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     2461 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     2521 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     2581 gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     2641 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     2701 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     2761 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     2821 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc
     2881 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg
     2941 gagcttgtat atccattttc ggatctgatc agcacgtgtt gacaattaat catcggcata
     3001 gtatatcggc atagtataat acgacaaggt gaggaactaa accatggcca agttgaccag
     3061 tgccgttccg gtgctcaccg cgcgcgacgt cgccggagcg gtcgagttct ggaccgaccg
     3121 gctcgggttc tcccgggact tcgtggagga cgacttcgcc ggtgtggtcc gggacgacgt
     3181 gaccctgttc atcagcgcgg tccaggacca ggtggtgccg gacaacaccc tggcctgggt
     3241 gtgggtgcgc ggcctggacg agctgtacgc cgagtggtcg gaggtcgtgt ccacgaactt
     3301 ccgggacgcc tccgggccgg ccatgaccga gatcggcgag cagccgtggg ggcgggagtt
     3361 cgccctgcgc gacccggccg gcaactgcgt gcacttcgtg gccgaggagc aggactgaca
     3421 cgtgctacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
     3481 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     3541 ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     3601 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     3661 tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc
     3721 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     3781 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taacttacat taattgcgtt
     3841 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     3901 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     3961 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     4021 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     4081 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     4141 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     4201 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     4261 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     4321 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     4381 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     4441 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     4501 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     4561 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     4621 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     4681 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     4741 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     4801 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     4861 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     4921 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     4981 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     5041 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     5101 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     5161 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     5221 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     5281 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     5341 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     5401 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     5461 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     5521 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     5581 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     5641 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     5701 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     5761 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     5821 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgc
//
