LOCUS       sequence-366777-        5594 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For the mammalian expression of the tardigrade protein CAHS1_HYPDU. 
            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 5594)
  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 5594)
  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_CAHS1_HYPDU (pBS0650)
FEATURES             Location/Qualifiers
     source          1..5594
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(55..74)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        246..625
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        626..829
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     780..800
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     874..893
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        874..892
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1613..1630)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1619..1843
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1889..2317
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1976..1995)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2186..2207
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2326..2346)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2331..2660
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2511..2646
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2573..2592
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        2708..2755
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2774..3148
                     /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    3278..3399
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3315..3334)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3369..3388
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3448..3464)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3448..3464)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3461..3483)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3472..3488
                     /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(3496..3502,3503..3520,3521..3526))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(3679..3696)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3850..4438)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3930..3949)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4609..5400,5401..5469))
                     /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     5232..5251
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5470..5574)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cacctgacgt cgacggatcg ggagatctcc cgatccccta tggtgcactc tcagtacaat
       61 ctgctctgat gccgcatagt taagccagta tctgctccct gcttgtgtgt tggaggtcgc
      121 tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg acaattgcat
      181 gaagaatctg cttagggtta ggcgttttgc gctgcttcgc gatgtacggg ccagatatac
      241 gcgttgacat tgattattga ctagttatta atagtaatca attacggggt cattagttca
      301 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc
      361 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
      421 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
      481 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc
      541 cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta
      601 cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg
      661 atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt
      721 gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac
      781 gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctc tctggctaac
      841 tagagaaccc actgcttact ggcttatcga aattaatacg actcactata gggagaccca
      901 agctggctag ccaccatgtc cggacggaat gtcgaatctc atatggaacg aaatgagaaa
      961 gtcgttgtca ataattcagg gcatgctgat gtcaagaaac aacaacaaca agtcgaacat
     1021 actgaattta ctcatacgga agtcaaagcg cctttgatac atccagcgcc acctataata
     1081 agtacaggcg cagccggact tgcagaagaa attgttggac aaggttttac agccagtgcc
     1141 gcgcgcatta gtggcggtac ggcggaagta catttgcaac cttcagcagc gatgactgaa
     1201 gaagcgcgtc gcgatcaaga aagatatcgg caagaacaag aatcaattgc taaacaacaa
     1261 gaacgagaaa tggaaaagaa aactgaagca tatcgcaaaa ctgctgaagc agaagcagaa
     1321 aagattcgca aagaacttga gaaacaacat gctagagatg tcgaatttcg gaaagatctt
     1381 attgaatcaa cgattgatcg tcaaaagcgg gaagtagatt tggaagctaa aatggctaaa
     1441 cgtgaattgg atagagaagg tcaacttgca aaagaagcgc tcgaaaggag tcgcttggct
     1501 acaaatgtcg aagtcaattt tgattctgca gcaggtcata cagttagtgg cggtacaaca
     1561 atatcatcaa gcgataaaat ggaaattaaa agaaattgaa cccgctgatc agcctcgact
     1621 gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg
     1681 gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg
     1741 agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg
     1801 gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga
     1861 accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg
     1921 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     1981 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     2041 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     2101 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     2161 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     2221 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     2281 aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt
     2341 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     2401 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     2461 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     2521 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     2581 cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg
     2641 gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc
     2701 agcacgtgtt gacaattaat catcggcata gtatatcggc atagtataat acgacaaggt
     2761 gaggaactaa accatggcca agttgaccag tgccgttccg gtgctcaccg cgcgcgacgt
     2821 cgccggagcg gtcgagttct ggaccgaccg gctcgggttc tcccgggact tcgtggagga
     2881 cgacttcgcc ggtgtggtcc gggacgacgt gaccctgttc atcagcgcgg tccaggacca
     2941 ggtggtgccg gacaacaccc tggcctgggt gtgggtgcgc ggcctggacg agctgtacgc
     3001 cgagtggtcg gaggtcgtgt ccacgaactt ccgggacgcc tccgggccgg ccatgaccga
     3061 gatcggcgag cagccgtggg ggcgggagtt cgccctgcgc gacccggccg gcaactgcgt
     3121 gcacttcgtg gccgaggagc aggactgaca cgtgctacga gatttcgatt ccaccgccgc
     3181 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     3241 gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa
     3301 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     3361 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac
     3421 ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     3481 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     3541 atgagtgagc taacttacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     3601 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     3661 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     3721 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     3781 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     3841 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     3901 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     3961 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     4021 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     4081 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     4141 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     4201 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     4261 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     4321 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     4381 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     4441 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     4501 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     4561 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     4621 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     4681 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     4741 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     4801 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     4861 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     4921 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     4981 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     5041 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     5101 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     5161 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     5221 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     5281 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     5341 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     5401 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     5461 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     5521 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     5581 tccccgaaaa gtgc
//
