LOCUS       sequence-366737-        5210 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For the mammalian expression of the tardigrade protein 
            CAHS2_RAMVA_94-170. This is one of the top performing targets in our
            apoptosis assay (it protected from apoptosis)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5210)
  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 5210)
  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_CAHS2_RAMVA_94-170 (pBS0295)
FEATURES             Location/Qualifiers
     source          1..5210
                     /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"
     regulatory      893..902
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1145..1168
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(1212..1229)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1218..1442
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1488..1916
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1575..1594)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1785..1806
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1925..1945)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1930..2259
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2110..2245
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2172..2191
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        2307..2354
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2373..2747
                     /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    2877..2998
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2914..2933)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2968..2987
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3047..3063)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3047..3063)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3060..3082)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3071..3087
                     /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(3095..3101,3102..3119,3120..3125))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(3278..3295)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3449..4037)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3529..3548)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4208..4999,5000..5068))
                     /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     4831..4850
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5069..5173)
                     /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 ggagagagag ctgcaggcca tcaacgagag acaccagaga gacatcgagg gcaagaccga
      961 ggcctacaga aagcaggccg agcaggaggc cgagagactg agaaaggagc tggagaagca
     1021 gcaccagaga gacatcgagt tcagaaagag cctggtgcag ggcaccatcg agaaccagaa
     1081 gagacaggtg gagctggagg cccagctggc caagagagag ctggacagag agtgaggatc
     1141 tggggactac aaggacgacg acgacaagtg aattgccaat tgaaatttga cggcctatac
     1201 ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc
     1261 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga
     1321 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga
     1381 cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat
     1441 ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag
     1501 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     1561 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     1621 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     1681 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     1741 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     1801 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     1861 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt
     1921 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     1981 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     2041 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     2101 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     2161 ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag
     2221 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata
     2281 tccattttcg gatctgatca gcacgtgttg acaattaatc atcggcatag tatatcggca
     2341 tagtataata cgacaaggtg aggaactaaa ccatggccaa gttgaccagt gccgttccgg
     2401 tgctcaccgc gcgcgacgtc gccggagcgg tcgagttctg gaccgaccgg ctcgggttct
     2461 cccgggactt cgtggaggac gacttcgccg gtgtggtccg ggacgacgtg accctgttca
     2521 tcagcgcggt ccaggaccag gtggtgccgg acaacaccct ggcctgggtg tgggtgcgcg
     2581 gcctggacga gctgtacgcc gagtggtcgg aggtcgtgtc cacgaacttc cgggacgcct
     2641 ccgggccggc catgaccgag atcggcgagc agccgtgggg gcgggagttc gccctgcgcg
     2701 acccggccgg caactgcgtg cacttcgtgg ccgaggagca ggactgacac gtgctacgag
     2761 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
     2821 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact
     2881 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     2941 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     3001 atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc
     3061 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     3121 gtaaagcctg gggtgcctaa tgagtgagct aacttacatt aattgcgttg cgctcactgc
     3181 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     3241 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     3301 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     3361 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     3421 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     3481 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     3541 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     3601 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     3661 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     3721 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     3781 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     3841 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     3901 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     3961 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     4021 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     4081 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     4141 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     4201 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     4261 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     4321 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     4381 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     4441 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     4501 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     4561 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     4621 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     4681 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     4741 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     4801 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     4861 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     4921 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     4981 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     5041 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     5101 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     5161 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga
//
