LOCUS       sequence-366782-        5363 bp    DNA     circular SYN 15-APR-2025
DEFINITION  For the mammalian expression of the tardigrade protein 
            CAHS1_PARRC_101-189_noLink. 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 5363)
  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 5363)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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_PARRC_101-189_noLink (pBS0653)
FEATURES             Location/Qualifiers
     source          1..5363
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(40..59)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        231..610
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        611..814
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     765..785
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     859..878
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        859..877
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      895..904
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(1367..1384)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1373..1597
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1643..2071
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1730..1749)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1940..1961
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2080..2100)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2085..2414
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2265..2400
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2327..2346
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        2462..2509
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2528..2902
                     /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    3032..3153
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3069..3088)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3123..3142
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3202..3218)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3202..3218)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3215..3237)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3226..3242
                     /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(3250..3256,3257..3274,3275..3280))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(3433..3450)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3604..4192)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3684..3703)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4363..5154,5155..5223))
                     /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     4986..5005
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5224..5328)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gatcgggaga tctcccgatc ccctatggtg cactctcagt acaatctgct ctgatgccgc
       61 atagttaagc cagtatctgc tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag
      121 caaaatttaa gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga atctgcttag
      181 ggttaggcgt tttgcgctgc ttcgcgatgt acgggccaga tatacgcgtt gacattgatt
      241 attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga
      301 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
      361 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
      421 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
      481 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
      541 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
      601 tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc
      661 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa
      721 tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag
      781 gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag aacccactgc
      841 ttactggctt atcgaaatta atacgactca ctatagggag acccaagctg gctagccacc
      901 atggccaaaa cgcgcgaaca ggagatgctg tctgctaact atcaaaaaga agttgagcgc
      961 aaaactgaag cctatcggaa gcagcaagaa gtggaggcag acaaaattcg gaaagagctg
     1021 gaaaagcagc atctgcgtaa agatattgta gagatggcaa tcgaaaacca gaaaaaaatg
     1081 attgatgtcg aaagccgtta tgcaaaaaaa gatatggacc gcgaacgtgt gaaagtccgt
     1141 atgatgctcg agcagtgatg aattgccaat tgaaatttga cggcctatgt cactgaaact
     1201 atccaaacac cccatgtcga tactgaacgt atcgacgcat acctccttcc ttgaaaacgc
     1261 acaatcatac aactgggcac ataatgcgta cgcccatcta gtacacccat ctctgtaggt
     1321 ccagttcaag agctggaaga gcaccctcca cttacccgct gatcagcctc gactgtgcct
     1381 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
     1441 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg
     1501 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac
     1561 aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc
     1621 tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     1681 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     1741 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     1801 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     1861 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     1921 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     1981 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     2041 gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt
     2101 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     2161 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     2221 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     2281 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     2341 ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     2401 taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct gatcagcacg
     2461 tgttgacaat taatcatcgg catagtatat cggcatagta taatacgaca aggtgaggaa
     2521 ctaaaccatg gccaagttga ccagtgccgt tccggtgctc accgcgcgcg acgtcgccgg
     2581 agcggtcgag ttctggaccg accggctcgg gttctcccgg gacttcgtgg aggacgactt
     2641 cgccggtgtg gtccgggacg acgtgaccct gttcatcagc gcggtccagg accaggtggt
     2701 gccggacaac accctggcct gggtgtgggt gcgcggcctg gacgagctgt acgccgagtg
     2761 gtcggaggtc gtgtccacga acttccggga cgcctccggg ccggccatga ccgagatcgg
     2821 cgagcagccg tgggggcggg agttcgccct gcgcgacccg gccggcaact gcgtgcactt
     2881 cgtggccgag gagcaggact gacacgtgct acgagatttc gattccaccg ccgccttcta
     2941 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     3001 ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta
     3061 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     3121 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag
     3181 ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     3241 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     3301 gagctaactt acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     3361 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     3421 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     3481 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     3541 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     3601 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     3661 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     3721 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     3781 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     3841 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     3901 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     3961 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     4021 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     4081 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     4141 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     4201 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     4261 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     4321 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     4381 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     4441 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     4501 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     4561 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     4621 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     4681 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     4741 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     4801 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     4861 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     4921 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     4981 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     5041 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     5101 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     5161 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     5221 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     5281 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     5341 aaaagtgcca cctgacgtcg acg
//
