LOCUS       sequence-363889-        7229 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For the mammalian expression of the synthetic protein 
            H2FLK5_CAEEL_30-800. This is one of the bottom performing targets in
            our apoptosis assay (it induced apoptosis)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7229)
  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 7229)
  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_H2FLK5_CAEEL_30-800 (pBS0864)
FEATURES             Location/Qualifiers
     source          1..7229
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(38..57)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        229..608
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        609..812
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     763..783
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     857..876
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        857..875
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3231..3248)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3237..3461
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3507..3935
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3594..3613)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3804..3825
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3944..3964)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3949..4278
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4129..4264
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4191..4210
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4326..4373
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4392..4766
                     /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    4896..5017
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4933..4952)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4987..5006
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5066..5082)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5066..5082)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5079..5101)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5090..5106
                     /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(5114..5120,5121..5138,5139..5144))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(5297..5314)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5468..6056)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5548..5567)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6227..7018,7019..7087))
                     /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     6850..6869
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7088..7192)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tcgggagatc tcccgatccc ctatggtgca ctctcagtac aatctgctct gatgccgcat
       61 agttaagcca gtatctgctc cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca
      121 aaatttaagc tacaacaagg caaggcttga ccgacaattg catgaagaat ctgcttaggg
      181 ttaggcgttt tgcgctgctt cgcgatgtac gggccagata tacgcgttga cattgattat
      241 tgactagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt
      301 tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc
      361 cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac
      421 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata
      481 tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc
      541 agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta
      601 ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac
      661 ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc
      721 aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc
      781 gtgtacggtg ggaggtctat ataagcagag ctctctggct aactagagaa cccactgctt
      841 actggcttat cgaaattaat acgactcact atagggagac ccaagctggc tagccaccat
      901 gaagaagttg gtggatgagg tgatacccgt ggcacaggga caggttggga accagcattt
      961 tgacgagcct aagtcccaga aaaccccgcc atctagcttg aatgacagct acgcagagga
     1021 gctgctcgag gcaaaattga ttaacaccga cgaatccagc aacactaccc acaataagat
     1081 gagctttatg gataaggtca aggacactgc caactctgct tacgagtccg tagaaaatac
     1141 agccgttggg gcaaaggacg gggtagtggg tgccttcaac actgtgggtg agaaaattgg
     1201 cggcgcctac gattctacaa aacaatatgc ttctgataaa gtgtctgccg ccggtgacta
     1261 tgcttcagag gcaggaaaga aggtcggaga cgccgaagac agcatgatac agggcgcgta
     1321 cgacgcaaag gatagcgcta aagaaagcgc ctctaatgcc gggaacaaga tttctaacgc
     1381 attcacctct ctgaaggact caatcaccgg cgacaacgcc cacaagactg gggaagccct
     1441 gaatgacaat tacgaaagtg tcaaggagag cgcgtcagag gctgccgata gacttaaaga
     1501 accagcacag aacacggcag acaacgccaa ggacttggca aaccagaccg gcgaggagat
     1561 gtccgatgca tttgagagcg tcaagcagct gacaagtgat acagctgaga acgccaagga
     1621 aaacgcccac gacgcggccg acaccgccaa atcttacaaa gacaaagccg gggagaaagc
     1681 ctctgggata cttgactctt tcaaagcaca cgcttccgac gcaaaggact ctgtcgaaaa
     1741 caaggcatca gacgcatata actccgccaa ggataaggcc ggtgacgcct gggattctac
     1801 gaaagacgct gcgggcgacg caaaagacaa gaccaaatcc ttcacagaaa aggtcgggga
     1861 taaaattagc ggagcctatg acagtgtcaa agagaaagcg agtgacgtgg ccgacagctt
     1921 caaggctcat agcacggaca gcaaggacaa cgtggagaat aaggcagccg acgcatataa
     1981 caccgccaag gataaggcct cagacgcatg ggacaaaaca aaggataagg caggcgaggc
     2041 gaaggataag atgggtgacg catgggacac caccaaagat aaagcagggg acgcctggga
     2101 cactaccaag gacaaggccg gagacggaaa gggtaaggca ggcgacgcct gggacacgac
     2161 caaagataaa gcatccgacg cctgggacac cacgaaggac aaggccggtg aggctaaaga
     2221 taagatgggc gaggcttggg accataccaa ggataaagca ggcgaggcta aggacaaggc
     2281 ttccgacgct gcagatgacg ctcagggcaa aagtaagagc atgaccgaaa agattggtga
     2341 ttcaatctct ggcgcttggg agagtaccaa agataccgca gtcagcgcta aggacaagac
     2401 cgctgaggct gcaggaaaag tcggtgatag tatttccggc gcttacgaca ccgtcaagga
     2461 gaaagcttct gatatcgccg actctttcaa ggcccacagt accgactcca aagacaacgt
     2521 tgagaacaag gccgcggacg cctacaactc tgccaaggac aaagccagtg acgcgtggga
     2581 caaaacaaaa gacaaggctg gcgacgctaa agataaagct gcagacgcct gggacacaac
     2641 taaggacaaa gccggtgacg cctgggactc cacgaaagac cacgcagctg acgcaaaaga
     2701 caaggccagc gatgcagccg gggacgcgaa agataaatct aagtccctca ctgaaaaggc
     2761 gggagacgca atatcaggcg catacgactc tgtgaaggag aaagcatctg acatcgctga
     2821 ctcatttaag gcccactcca ccaacagcaa agacaacgtg gaaaacaaag caagcgacgc
     2881 gtacaactcc gccaaggaca aggcatccga cgcttgggac aaaacgaaag ataaggccgg
     2941 tgaggctaaa gacaaagctg gcgacgcttg ggataatacc aaggataaag ctgggaacgc
     3001 atgggactca acaaaggata aagcatccga cgcctgggac accacaaaag ataaggcgtc
     3061 agacgccaaa gagtctgcgg gcgacgctgc agacagtgcc aaagacaaga gtaaaagcat
     3121 tacggagacc atcggcgaca agataagcgg agcctacgat tccgtcaagg aaaaggcttc
     3181 cgacgtggcc gattccttta aagcgcatag ctcttagacc cgctgatcag cctcgactgt
     3241 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
     3301 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
     3361 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga
     3421 agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac
     3481 cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg
     3541 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     3601 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     3661 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     3721 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     3781 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     3841 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     3901 aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta
     3961 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     4021 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     4081 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     4141 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     4201 gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     4261 ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcag
     4321 cacgtgttga caattaatca tcggcatagt atatcggcat agtataatac gacaaggtga
     4381 ggaactaaac catggccaag ttgaccagtg ccgttccggt gctcaccgcg cgcgacgtcg
     4441 ccggagcggt cgagttctgg accgaccggc tcgggttctc ccgggacttc gtggaggacg
     4501 acttcgccgg tgtggtccgg gacgacgtga ccctgttcat cagcgcggtc caggaccagg
     4561 tggtgccgga caacaccctg gcctgggtgt gggtgcgcgg cctggacgag ctgtacgccg
     4621 agtggtcgga ggtcgtgtcc acgaacttcc gggacgcctc cgggccggcc atgaccgaga
     4681 tcggcgagca gccgtggggg cgggagttcg ccctgcgcga cccggccggc aactgcgtgc
     4741 acttcgtggc cgaggagcag gactgacacg tgctacgaga tttcgattcc accgccgcct
     4801 tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc
     4861 gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg
     4921 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     4981 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct
     5041 ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     5101 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     5161 gagtgagcta acttacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     5221 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     5281 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     5341 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     5401 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     5461 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     5521 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     5581 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     5641 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     5701 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     5761 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     5821 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     5881 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     5941 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     6001 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     6061 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     6121 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     6181 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     6241 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     6301 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     6361 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     6421 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     6481 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     6541 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     6601 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     6661 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     6721 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     6781 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     6841 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     6901 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     6961 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     7021 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     7081 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     7141 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     7201 cccgaaaagt gccacctgac gtcgacgga
//
