LOCUS       sequence-366865-        5216 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For the mammalian expression of the C. elegans protein 
            Q19228_CAEEL_93-127. 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 5216)
  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 5216)
  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_Q19228_CAEEL_93-127 (pBS0735)
FEATURES             Location/Qualifiers
     source          1..5216
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(43..62)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        234..613
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        614..817
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     768..788
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     862..881
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        862..880
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1223..1240)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1229..1453
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1499..1927
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1586..1605)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1796..1817
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1936..1956)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1941..2270
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2121..2256
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2183..2202
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        2318..2365
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2384..2758
                     /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    2888..3009
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2925..2944)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2979..2998
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3058..3074)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3058..3074)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3071..3093)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3082..3098
                     /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(3106..3112,3113..3130,3131..3136))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3151..3172
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3289..3306)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3460..4048)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3540..3559)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4219..5010,5011..5079))
                     /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     4842..4861
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5080..5184)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 acggatcggg agatctcccg atcccctatg gtcgactctc agtacaatct gctctgatgc
       61 cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc
      121 gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga agaatctgct
      181 tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc gttgacattg
      241 attattgact agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat
      301 ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc
      361 ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca
      421 ttgacgtcaa tgggtggact atttacggta aactgcccac ttggcagtac atcaagtgta
      481 tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta
      541 tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat
      601 cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga
      661 ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca
      721 aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg
      781 taggcgtgta cggtgggagg tctatataag cagagctctc tggctaacta gagaacccac
      841 tgcttactgg cttatcgaaa ttaatacgac tcactatagg gagacccaag ctggctagcc
      901 accatgtcat tttctcgcat gatcctgctg cctgaggacg tggacatcac cagcgtggcc
      961 actaacctgt ctaacgacgg caagctgtgc atcgaggctc ccaagctgga gtgatgaatt
     1021 gccaattgaa atttgacggc ctatgtcact gaaactatcc aaacacccca tgtcgatact
     1081 gaacgtatcg acgcatacct ccttccttga aaacgcacaa tcatacaact gggcacataa
     1141 tgcgtacgcc catctagtac acccatctct gtaggtccag ttcaagagct ggaagagcac
     1201 cctccactta cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt
     1261 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
     1321 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
     1381 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg
     1441 gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac
     1501 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     1561 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     1621 ttcgccggct ttccccgtca agctctaaat cggggcatcc ctttagggtt ccgatttagt
     1681 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     1741 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     1801 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     1861 gggattttgg ggatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     1921 gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     1981 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     2041 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     2101 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     2161 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc
     2221 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg
     2281 gagcttgtat atccattttc ggatctgatc agcacgtgtt gacaattaat catcggcata
     2341 gtatatcggc atagtataat acgacaaggt gaggaactaa accatggcca agttgaccag
     2401 tgccgttccg gtgctcaccg cgcgcgacgt cgccggagcg gtcgagttct ggaccgaccg
     2461 gctcgggttc tcccgggact tcgtggagga cgacttcgcc ggtgtggtcc gggacgacgt
     2521 gaccctgttc atcagcgcgg tccaggacca ggtggtgccg gacaacaccc tggcctgggt
     2581 gtgggtgcgc ggcctggacg agctgtacgc cgagtggtcg gaggtcgtgt ccacgaactt
     2641 ccgggacgcc tccgggccgg ccatgaccga gatcggcgag cagccgtggg ggcgggagtt
     2701 cgccctgcgc gacccggccg gcaactgcgt gcacttcgtg gccgaggagc aggactgaca
     2761 cgtgctacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
     2821 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     2881 ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     2941 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     3001 tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc
     3061 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     3121 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     3181 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     3241 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     3301 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     3361 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     3421 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     3481 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     3541 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     3601 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc
     3661 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     3721 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     3781 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     3841 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     3901 gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     3961 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     4021 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     4081 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     4141 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     4201 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     4261 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     4321 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     4381 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     4441 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     4501 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     4561 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     4621 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     4681 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     4741 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     4801 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     4861 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     4921 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     4981 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     5041 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     5101 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     5161 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcg
//
