LOCUS       sequence-366844-        5693 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For the mammalian expression of the C. elegans protein 
            H2FLK5_CAEEL_542-800. 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 5693)
  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 5693)
  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_542-800 (pBS0695)
FEATURES             Location/Qualifiers
     source          1..5693
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(46..65)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        237..616
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        617..820
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     771..791
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     865..884
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        865..883
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      901..910
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(1703..1720)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1709..1933
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1979..2407
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2066..2085)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2276..2297
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2416..2436)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2421..2750
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2601..2736
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2663..2682
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        2798..2845
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2864..3238
                     /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    3368..3489
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3405..3424)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3459..3478
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3538..3554)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3538..3554)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3551..3573)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3562..3578
                     /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(3586..3592,3593..3610,3611..3616))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(3769..3786)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3940..4528)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4020..4039)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4699..5490,5491..5559))
                     /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     5322..5341
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5560..5664)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga
       61 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg
      121 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct
      181 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca
      241 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata
      301 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
      361 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
      421 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
      481 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
      541 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
      601 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
      661 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
      721 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
      781 cggtaggcgt gtacggtggg aggtctatat aagcagagct ctctggctaa ctagagaacc
      841 cactgcttac tggcttatcg aaattaatac gactcactat agggagaccc aagctggcta
      901 gccaccatgg gggcatatga tacagtgaaa gaaaaggcat cagacattgc agattcattt
      961 aaagcgcatt ccacagattc aaaggataat gtcgaaaata aagcggctga tgcttataat
     1021 agtgcgaaag ataaggcaag cgatgcttgg gataagacta aggataaagc aggagatgcg
     1081 aaggacaagg ccgctgatgc atgggatacc acgaaagata aggcaggcga tgcttgggat
     1141 agtacaaagg atcatgccgc agatgctaag gataaagctt cagacgctgc tggcgatgca
     1201 aaggacaagt ccaaatcact gaccgagaaa gctggcgatg ctatttccgg agcgtatgat
     1261 tcagtcaaag aaaaggccag cgatattgcg gatagcttca aagcacattc tacgaatagt
     1321 aaggataatg ttgagaataa ggcgagtgat gcatataata gtgcaaaaga taaagcatct
     1381 gatgcatggg ataagactaa ggacaaagca ggcgaagcca aggataaggc cggagatgcc
     1441 tgggacaaca ctaaagacaa ggcaggaaat gcgtgggata gtactaaaga caaggcttct
     1501 gatgcatggg atacaaccaa ggacaaagcc agtgatgcta aggaaagcgc tggagatgcc
     1561 gctgattctg ctaaggataa atccaagtca ataactgaaa caattgggga taaaatctcc
     1621 ggcgcttatg actctgttaa agagaaagcc agcgatgttg ctgactcatt caaggctcac
     1681 tccagttgaa cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt
     1741 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
     1801 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
     1861 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg
     1921 gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac
     1981 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     2041 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     2101 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     2161 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     2221 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     2281 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     2341 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     2401 gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     2461 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     2521 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     2581 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     2641 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc
     2701 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg
     2761 gagcttgtat atccattttc ggatctgatc agcacgtgtt gacaattaat catcggcata
     2821 gtatatcggc atagtataat acgacaaggt gaggaactaa accatggcca agttgaccag
     2881 tgccgttccg gtgctcaccg cgcgcgacgt cgccggagcg gtcgagttct ggaccgaccg
     2941 gctcgggttc tcccgggact tcgtggagga cgacttcgcc ggtgtggtcc gggacgacgt
     3001 gaccctgttc atcagcgcgg tccaggacca ggtggtgccg gacaacaccc tggcctgggt
     3061 gtgggtgcgc ggcctggacg agctgtacgc cgagtggtcg gaggtcgtgt ccacgaactt
     3121 ccgggacgcc tccgggccgg ccatgaccga gatcggcgag cagccgtggg ggcgggagtt
     3181 cgccctgcgc gacccggccg gcaactgcgt gcacttcgtg gccgaggagc aggactgaca
     3241 cgtgctacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
     3301 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     3361 ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     3421 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     3481 tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc
     3541 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     3601 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taacttacat taattgcgtt
     3661 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     3721 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     3781 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     3841 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     3901 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     3961 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     4021 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     4081 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     4141 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     4201 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     4261 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     4321 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     4381 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     4441 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     4501 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     4561 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     4621 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     4681 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     4741 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     4801 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     4861 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     4921 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     4981 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     5041 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     5101 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     5161 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     5221 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     5281 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     5341 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     5401 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     5461 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     5521 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     5581 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     5641 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acg
//
