LOCUS       sequence-366845-        5627 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For the mammalian expression of the C. elegans protein 
            H2FLK5_CAEEL_199-435. 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 5627)
  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 5627)
  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_199-435 (pBS0696)
FEATURES             Location/Qualifiers
     source          1..5627
                     /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(1637..1654)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1643..1867
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1913..2341
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2000..2019)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2210..2231
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2350..2370)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2355..2684
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2535..2670
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2597..2616
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        2732..2779
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2798..3172
                     /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    3302..3423
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3339..3358)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3393..3412
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3472..3488)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3472..3488)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3485..3507)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3496..3512
                     /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(3520..3526,3527..3544,3545..3550))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(3703..3720)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3874..4462)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3954..3973)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4633..5424,5425..5493))
                     /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     5256..5275
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5494..5598)
                     /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 gagataatgc gcataaaacg ggagaagctt tgaacgataa ctatgagagc
      961 gtgaaagaaa gtgccagcga agcagcagac cgattgaagg agcccgccca aaatacagcg
     1021 gataatgcaa aagatctcgc taatcaaact ggagaagaaa tgagtgacgc gttcgaatca
     1081 gtgaaacaat tgacttccga cacggccgaa aatgcgaaag agaatgcaca tgatgcagct
     1141 gatacagcaa agtcatataa ggataaggca ggagaaaagg ctagtggaat tctggatagt
     1201 tttaaggccc atgcgtctga tgcgaaagat tccgtggaga ataaagctag cgatgcctac
     1261 aattctgcaa aagacaaagc tggagatgct tgggactcaa caaaggatgc cgccggagat
     1321 gcgaaggata aaacaaagag ttttactgag aaagtgggcg acaagatatc cggggcttac
     1381 gattccgtga aggaaaaggc atccgatgtt gcagattcat ttaaagccca ctcaactgat
     1441 tccaaagata atgtcgaaaa caaagctgca gatgcgtaca atactgcaaa agacaaagct
     1501 agcgatgcct gggataagac gaaagacaaa gccggagaag ccaaagacaa aatgggagat
     1561 gcctgggata caacgaagga caaggctggc gatgcgtggg atacgacaaa agataaagcg
     1621 tgaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc
     1681 tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat
     1741 gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg
     1801 caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc
     1861 tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc
     1921 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     1981 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     2041 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     2101 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     2161 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     2221 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     2281 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     2341 taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     2401 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     2461 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     2521 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     2581 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc
     2641 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt
     2701 gtatatccat tttcggatct gatcagcacg tgttgacaat taatcatcgg catagtatat
     2761 cggcatagta taatacgaca aggtgaggaa ctaaaccatg gccaagttga ccagtgccgt
     2821 tccggtgctc accgcgcgcg acgtcgccgg agcggtcgag ttctggaccg accggctcgg
     2881 gttctcccgg gacttcgtgg aggacgactt cgccggtgtg gtccgggacg acgtgaccct
     2941 gttcatcagc gcggtccagg accaggtggt gccggacaac accctggcct gggtgtgggt
     3001 gcgcggcctg gacgagctgt acgccgagtg gtcggaggtc gtgtccacga acttccggga
     3061 cgcctccggg ccggccatga ccgagatcgg cgagcagccg tgggggcggg agttcgccct
     3121 gcgcgacccg gccggcaact gcgtgcactt cgtggccgag gagcaggact gacacgtgct
     3181 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
     3241 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
     3301 caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     3361 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     3421 ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct
     3481 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     3541 aaagtgtaaa gcctggggtg cctaatgagt gagctaactt acattaattg cgttgcgctc
     3601 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     3661 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     3721 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     3781 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     3841 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     3901 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     3961 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     4021 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     4081 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     4141 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     4201 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     4261 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     4321 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     4381 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     4441 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     4501 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     4561 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     4621 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     4681 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     4741 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     4801 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     4861 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     4921 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     4981 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     5041 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     5101 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     5161 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     5221 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     5281 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     5341 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     5401 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     5461 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     5521 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     5581 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacg
//
