LOCUS       sequence-366774-        5864 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For the mammalian expression of the human protein APOE_HUMAN_E3. 
            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_E3 (pBS0648)
FEATURES             Location/Qualifiers
     source          1..5864
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1872..1889)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1878..2102
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2148..2576
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2235..2254)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2445..2466
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2585..2605)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2590..2919
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2770..2905
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2832..2851
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        2967..3014
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3033..3407
                     /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    3537..3658
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3574..3593)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3628..3647
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3707..3723)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3707..3723)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3720..3742)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3731..3747
                     /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(3755..3761,3762..3779,3780..3785))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(3938..3955)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4109..4697)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4189..4208)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4868..5659,5660..5728))
                     /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     5491..5510
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5729..5833)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 caccatgaaa gtgttatggg cagcattgtt ggtgacattt ttagctgggt gccaggccaa
      961 agtggaacaa gcagtggaaa cagaacctga acctgaactg agacagcaga ccgaatggca
     1021 aagtggccag agatgggaac tggccctggg gagattttgg gactatctga gatgggtgca
     1081 gaccctgagc gagcaggtgc aggaggagct cctgagctct caggtgacac aggaactgag
     1141 agccctgatg gacgagacta tgaaggaact gaaggcctac aaatctgagc tggaagagca
     1201 gctgaccccc gtggctgaag agacccgcgc tcgcttgtcc aaagagctcc aggccgccca
     1261 ggcccggctg ggtgccgata tggaggacgt ctgcggccgc ctggtccagt accgcgggga
     1321 agtgcaggct atgctggggc agtctacaga ggagctccgg gtcagactgg cctctcacct
     1381 gcggaagctg aggaagaggt tgctccgcga cgccgacgac ctccagaagc gcctggctgt
     1441 gtaccaggcc ggcgcccggg aaggcgccga gcgcggactc tccgctatca gggagcggct
     1501 cggccctctg gttgaacagg gcagagtcag ggctgccact gtggggagcc tcgccggaca
     1561 gcccctccag gagagagccc aggcctgggg cgagcgcctg cgagctagaa tggaggagat
     1621 ggggtccaga acccgcgacc gcctggacga agtgaaggag caggtggccg aggtgcgtgc
     1681 caaactggag gaacaggccc agcagatccg cctccaggct gaggccttcc aggccagact
     1741 gaagagctgg tttgagcctc ttgtggagga tatgcagcgg cagtgggccg ggctggtcga
     1801 gaaggtgcag gccgccgtgg gcacttccgc cgcccctgtc cccagcgaca accattgaac
     1861 ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc
     1921 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga
     1981 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga
     2041 cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat
     2101 ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag
     2161 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     2221 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     2281 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     2341 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     2401 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     2461 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     2521 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt
     2581 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     2641 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     2701 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     2761 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     2821 ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag
     2881 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata
     2941 tccattttcg gatctgatca gcacgtgttg acaattaatc atcggcatag tatatcggca
     3001 tagtataata cgacaaggtg aggaactaaa ccatggccaa gttgaccagt gccgttccgg
     3061 tgctcaccgc gcgcgacgtc gccggagcgg tcgagttctg gaccgaccgg ctcgggttct
     3121 cccgggactt cgtggaggac gacttcgccg gtgtggtccg ggacgacgtg accctgttca
     3181 tcagcgcggt ccaggaccag gtggtgccgg acaacaccct ggcctgggtg tgggtgcgcg
     3241 gcctggacga gctgtacgcc gagtggtcgg aggtcgtgtc cacgaacttc cgggacgcct
     3301 ccgggccggc catgaccgag atcggcgagc agccgtgggg gcgggagttc gccctgcgcg
     3361 acccggccgg caactgcgtg cacttcgtgg ccgaggagca ggactgacac gtgctacgag
     3421 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
     3481 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact
     3541 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     3601 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     3661 atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc
     3721 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     3781 gtaaagcctg gggtgcctaa tgagtgagct aacttacatt aattgcgttg cgctcactgc
     3841 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     3901 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     3961 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     4021 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     4081 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     4141 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     4201 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     4261 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     4321 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     4381 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     4441 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     4501 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     4561 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     4621 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     4681 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     4741 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     4801 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     4861 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     4921 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     4981 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     5041 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     5101 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     5161 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     5221 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     5281 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     5341 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     5401 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     5461 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     5521 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     5581 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     5641 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     5701 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     5761 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     5821 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtc
//
