LOCUS       sequence-363815-        5300 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For the mammalian expression of the human protein 
            APOE_HUMAN_E2_255-317. 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 5300)
  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 5300)
  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_E2_255-317 (pBS0848)
FEATURES             Location/Qualifiers
     source          1..5300
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(47..66)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        238..617
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        618..821
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     772..792
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     866..885
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        866..884
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      902..911
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(1311..1328)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1317..1541
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1587..2015
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1674..1693)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1884..1905
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2024..2044)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2029..2358
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2209..2344
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2271..2290
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        2406..2453
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2472..2846
                     /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    2976..3097
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3013..3032)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3067..3086
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3146..3162)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3146..3162)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3159..3181)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3170..3186
                     /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(3194..3200,3201..3218,3219..3224))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(3377..3394)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3548..4136)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3628..3647)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4307..5098,5099..5167))
                     /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     4930..4949
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5168..5272)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
       61 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
      121 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
      181 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac
      241 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
      301 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
      361 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      421 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      481 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      541 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
      601 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
      661 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
      721 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
      781 gcggtaggcg tgtacggtgg gaggtctata taagcagagc tctctggcta actagagaac
      841 ccactgctta ctggcttatc gaaattaata cgactcacta tagggagacc caagctggct
      901 agccaccatg gccgaggtgc gtgccaaact ggaggaacag gcccagcaga tccgcctcca
      961 ggctgaggcc ttccaggcca gactgaagag ctggtttgag cctcttgtgg aggatatgca
     1021 gcggcagtgg gccgggctgg tcgagaaggt gcaggccgcc gtgggcactt ccgccgcccc
     1081 tgtccccagc gacaaccatt gatgaattgc caattgaaat ttgacggcct atgtcactga
     1141 aactatccaa acaccccatg tcgatactga acgtatcgac gcatacctcc ttccttgaaa
     1201 acgcacaatc atacaactgg gcacataatg cgtacgccca tctagtacac ccatctctgt
     1261 aggtccagtt caagagctgg aagagcaccc tccacttacc cgctgatcag cctcgactgt
     1321 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
     1381 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
     1441 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga
     1501 agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac
     1561 cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg
     1621 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     1681 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     1741 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     1801 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     1861 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     1921 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     1981 aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta
     2041 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     2101 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     2161 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     2221 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     2281 gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     2341 ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcag
     2401 cacgtgttga caattaatca tcggcatagt atatcggcat agtataatac gacaaggtga
     2461 ggaactaaac catggccaag ttgaccagtg ccgttccggt gctcaccgcg cgcgacgtcg
     2521 ccggagcggt cgagttctgg accgaccggc tcgggttctc ccgggacttc gtggaggacg
     2581 acttcgccgg tgtggtccgg gacgacgtga ccctgttcat cagcgcggtc caggaccagg
     2641 tggtgccgga caacaccctg gcctgggtgt gggtgcgcgg cctggacgag ctgtacgccg
     2701 agtggtcgga ggtcgtgtcc acgaacttcc gggacgcctc cgggccggcc atgaccgaga
     2761 tcggcgagca gccgtggggg cgggagttcg ccctgcgcga cccggccggc aactgcgtgc
     2821 acttcgtggc cgaggagcag gactgacacg tgctacgaga tttcgattcc accgccgcct
     2881 tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc
     2941 gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg
     3001 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     3061 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct
     3121 ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     3181 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     3241 gagtgagcta acttacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     3301 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     3361 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     3421 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     3481 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     3541 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     3601 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     3661 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     3721 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     3781 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     3841 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     3901 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     3961 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     4021 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     4081 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     4141 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     4201 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     4261 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     4321 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     4381 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     4441 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     4501 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     4561 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     4621 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     4681 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     4741 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     4801 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     4861 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     4921 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     4981 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     5041 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     5101 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     5161 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     5221 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     5281 cccgaaaagt gccacctgac
//
