LOCUS       sequence-363805-        5864 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For the mammalian expression of the human protein APOE_HUMAN_E114I. 
            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_E114I (pBS0839)
FEATURES             Location/Qualifiers
     source          1..5864
                     /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"
     primer_bind     878..897
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        878..896
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1887..1904)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1893..2117
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2163..2591
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2250..2269)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2460..2481
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2600..2620)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2605..2934
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2785..2920
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2847..2866
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        2982..3029
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3048..3422
                     /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    3552..3673
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3589..3608)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3643..3662
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3722..3738)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3722..3738)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3735..3757)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3746..3762
                     /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(3770..3776,3777..3794,3795..3800))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(3953..3970)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4124..4712)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4204..4223)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4883..5674,5675..5743))
                     /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     5506..5525
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5744..5848)
                     /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 gctctctggc
      841 taactagaga acccactgct tactggctta tcgaaattaa tacgactcac tatagggaga
      901 cccaagctgg ctagccacca tgaaagtgtt atgggcagca ttgttggtga catttttagc
      961 tgggtgccag gccaaagtgg aacaagcagt ggaaacagaa cctgaacctg aactgagaca
     1021 gcagaccgaa tggcaaagtg gccagagatg ggaactggcc ctggggagat tttgggacta
     1081 tctgagatgg gtgcagaccc tgagcgagca ggtgcaggag gagctcctga gctctcaggt
     1141 gacacaggaa ctgagagccc tgatggacga gactatgaag gaactgaagg cctacaaatc
     1201 tgagctggaa gagcagctga cccccgtggc tgaagagacc cgcgctcgct tgtccaaaat
     1261 cctccaggcc gcccaggccc ggctgggtgc cgatatggag gacgtctgcg gccgcctggt
     1321 ccagtaccgc ggggaagtgc aggctatgct ggggcagtct acagaggagc tccgggtcag
     1381 actggcctct cacctgcgga agctgaggaa gaggttgctc cgcgacgccg acgacctcca
     1441 gaagcgcctg gctgtgtacc aggccggcgc ccgggaaggc gccgagcgcg gactctccgc
     1501 tatcagggag cggctcggcc ctctggttga acagggcaga gtcagggctg ccactgtggg
     1561 gagcctcgcc ggacagcccc tccaggagag agcccaggcc tggggcgagc gcctgcgagc
     1621 tagaatggag gagatggggt ccagaacccg cgaccgcctg gacgaagtga aggagcaggt
     1681 ggccgaggtg cgtgccaaac tggaggaaca ggcccagcag atccgcctcc aggctgaggc
     1741 cttccaggcc agactgaaga gctggtttga gcctcttgtg gaggatatgc agcggcagtg
     1801 ggccgggctg gtcgagaagg tgcaggccgc cgtgggcact tccgccgccc ctgtccccag
     1861 cgacaaccat tgaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt
     1921 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc
     1981 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg
     2041 tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga
     2101 tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc
     2161 ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     2221 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     2281 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     2341 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     2401 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     2461 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     2521 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     2581 taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     2641 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     2701 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     2761 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     2821 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct
     2881 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc
     2941 ccgggagctt gtatatccat tttcggatct gatcagcacg tgttgacaat taatcatcgg
     3001 catagtatat cggcatagta taatacgaca aggtgaggaa ctaaaccatg gccaagttga
     3061 ccagtgccgt tccggtgctc accgcgcgcg acgtcgccgg agcggtcgag ttctggaccg
     3121 accggctcgg gttctcccgg gacttcgtgg aggacgactt cgccggtgtg gtccgggacg
     3181 acgtgaccct gttcatcagc gcggtccagg accaggtggt gccggacaac accctggcct
     3241 gggtgtgggt gcgcggcctg gacgagctgt acgccgagtg gtcggaggtc gtgtccacga
     3301 acttccggga cgcctccggg ccggccatga ccgagatcgg cgagcagccg tgggggcggg
     3361 agttcgccct gcgcgacccg gccggcaact gcgtgcactt cgtggccgag gagcaggact
     3421 gacacgtgct acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
     3481 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
     3541 tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     3601 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     3661 tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat
     3721 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     3781 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactt acattaattg
     3841 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     3901 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     3961 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     4021 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     4081 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     4141 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     4201 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     4261 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     4321 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     4381 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     4441 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     4501 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     4561 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     4621 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     4681 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     4741 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     4801 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     4861 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     4921 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     4981 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     5041 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     5101 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     5161 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     5221 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     5281 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     5341 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     5401 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     5461 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     5521 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     5581 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     5641 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     5701 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     5761 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     5821 agaaaaataa acaaataggg gttccgcgca catttccccg aaaa
//
