LOCUS       sequence-366641-        4563 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For the mammalian expression of the tardigrade protein CAHS1_PARRC. 
            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 4563)
  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 4563)
  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: CAHS1_PARRC_Boothby pSecTag2(pm) (pBS0258)
FEATURES             Location/Qualifiers
     source          1..4563
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(1..105)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(180..199)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        371..750
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        751..954
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     905..925
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     999..1018
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        999..1017
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1034..1043
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1043..1066
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        1802..1849
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             1868..2242
                     /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    2372..2493
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2409..2428)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2463..2482
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2542..2558)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2542..2558)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2555..2577)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2566..2582
                     /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(2590..2596,2597..2614,2615..2620))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(2773..2790)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2944..3532)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3024..3043)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3703..4494,4495..4563))
                     /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     4326..4345
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
       61 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
      121 agtgccacct gacgtcgacg gatcgggaga tctcccgatc ccctatggtg cactctcagt
      181 acaatctgct ctgatgccgc atagttaagc cagtatctgc tccctgcttg tgtgttggag
      241 gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt gaccgacaat
      301 tgcatgaaga atctgcttag ggttaggcgt tttgcgctgc ttcgcgatgt acgggccaga
      361 tatacgcgtt gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta
      421 gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc
      481 tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg
      541 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg
      601 gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa
      661 tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac
      721 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg
      781 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg
      841 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca
      901 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg
      961 ctaactagag aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag
     1021 acccgaattc gccgccacca tggattacaa ggatgatgat gataagatga tgtcagcgga
     1081 agctatgaac atgaacatga accaggatgc agtttttatt cctcctccgg aaggcgaaca
     1141 gtatgaacgc aaagagaaac aagagattca acagacgtcg tacctgcaga gccaggtaaa
     1201 ggttccgctg gtcaacctgc ccgcaccttt cttctcaacc tccttctccg cgcaggagat
     1261 ccttggggaa ggtttccagg ctagtatttc tcggatttcc gccgttagtg aagaactgtc
     1321 ctctatcgaa attccggagc tggcggaaga ggcccgtcgt gactttgcgg ccaaaacgcg
     1381 cgaacaggag atgctgtctg ctaactatca aaaagaagtt gagcgcaaaa ctgaagccta
     1441 tcggaagcag caagaagtgg aggcagacaa aattcggaaa gagctggaaa agcagcatct
     1501 gcgtgacgtg gagttccgca aagatattgt agagatggca atcgaaaacc agaaaaaaat
     1561 gattgatgtc gaaagccgtt atgcaaaaaa agatatggac cgcgaacgtg tgaaagtccg
     1621 tatgatgctc gagcagcaaa aatttcatag cgacatccag gtaaatctcg attctagcgc
     1681 tgcagggacc gaaactggag gccaagttgt ttcagaatct cagaaattta ccgaacgtaa
     1741 tcgtcagatt aaacagtgag ggtgtttgtg gttttgttgt gtggtattgt gatcagcacg
     1801 tgttgacaat taatcatcgg catagtatat cggcatagta taatacgaca aggtgaggaa
     1861 ctaaaccatg gccaagttga ccagtgccgt tccggtgctc accgcgcgcg acgtcgccgg
     1921 agcggtcgag ttctggaccg accggctcgg gttctcccgg gacttcgtgg aggacgactt
     1981 cgccggtgtg gtccgggacg acgtgaccct gttcatcagc gcggtccagg accaggtggt
     2041 gccggacaac accctggcct gggtgtgggt gcgcggcctg gacgagctgt acgccgagtg
     2101 gtcggaggtc gtgtccacga acttccggga cgcctccggg ccggccatga ccgagatcgg
     2161 cgagcagccg tgggggcggg agttcgccct gcgcgacccg gccggcaact gcgtgcactt
     2221 cgtggccgag gagcaggact gacacgtgct acgagatttc gattccaccg ccgccttcta
     2281 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     2341 ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta
     2401 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     2461 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag
     2521 ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     2581 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     2641 gagctaactt acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     2701 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     2761 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     2821 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     2881 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     2941 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     3001 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     3061 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     3121 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     3181 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     3241 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     3301 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     3361 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     3421 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     3481 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     3541 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     3601 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     3661 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     3721 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     3781 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     3841 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     3901 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     3961 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     4021 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     4081 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     4141 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     4201 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     4261 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     4321 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     4381 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     4441 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     4501 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     4561 cat
//
