LOCUS       sequence-359085-        4910 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  For the mammalian expression of the tardigrade protein CAHS6_HYPDU. 
            This is one of the top performing targets in our apoptosis assay (it
            protected from apoptosis)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 4910)
  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 4910)
  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_CAHS6_HYPDU (pBS0331)
FEATURES             Location/Qualifiers
     source          1..4910
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     complement(14..31)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    20..244
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      290..718
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(377..396)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     587..608
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(727..747)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        732..1061
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      912..1047
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     974..993
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        1109..1156
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             1175..1549
                     /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    1679..1800
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1716..1735)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1770..1789
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1849..1865)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1849..1865)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1862..1884)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1873..1889
                     /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(1897..1903,1904..1921,1922..1927))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1942..1963
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2080..2097)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2251..2839)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2331..2350)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3010..3801,3802..3870))
                     /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     3633..3652
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3871..3975)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4050..4069)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4241..4620
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4621..4824
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4775..4795
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4869..4888
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4869..4887
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 acccgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc
       61 cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag
      121 gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag
      181 gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct
      241 atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt
      301 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
      361 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
      421 tttccccgtc aagctctaaa tcggggcatc cctttagggt tccgatttag tgctttacgg
      481 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
      541 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
      601 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
      661 gggatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa
      721 ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
      781 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
      841 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
      901 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
      961 gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga
     1021 agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta
     1081 tatccatttt cggatctgat cagcacgtgt tgacaattaa tcatcggcat agtatatcgg
     1141 catagtataa tacgacaagg tgaggaacta aaccatggcc aagttgacca gtgccgttcc
     1201 ggtgctcacc gcgcgcgacg tcgccggagc ggtcgagttc tggaccgacc ggctcgggtt
     1261 ctcccgggac ttcgtggagg acgacttcgc cggtgtggtc cgggacgacg tgaccctgtt
     1321 catcagcgcg gtccaggacc aggtggtgcc ggacaacacc ctggcctggg tgtgggtgcg
     1381 cggcctggac gagctgtacg ccgagtggtc ggaggtcgtg tccacgaact tccgggacgc
     1441 ctccgggccg gccatgaccg agatcggcga gcagccgtgg gggcgggagt tcgccctgcg
     1501 cgacccggcc ggcaactgcg tgcacttcgt ggccgaggag caggactgac acgtgctacg
     1561 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
     1621 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa
     1681 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     1741 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     1801 tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt
     1861 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     1921 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     1981 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     2041 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     2101 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     2161 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     2221 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     2281 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     2341 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     2401 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcaatgct cacgctgtag
     2461 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     2521 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     2581 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     2641 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt
     2701 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     2761 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     2821 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     2881 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     2941 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     3001 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     3061 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     3121 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     3181 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     3241 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     3301 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     3361 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     3421 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     3481 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     3541 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     3601 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     3661 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     3721 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     3781 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     3841 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     3901 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     3961 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga
     4021 tctcccgatc ccctatggtc gactctcagt acaatctgct ctgatgccgc atagttaagc
     4081 cagtatctgc tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa
     4141 gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga atctgcttag ggttaggcgt
     4201 tttgcgctgc ttcgcgatgt acgggccaga tatacgcgtt gacattgatt attgactagt
     4261 tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt
     4321 acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg
     4381 tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg
     4441 gtggactatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt
     4501 acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg
     4561 accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg
     4621 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
     4681 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
     4741 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
     4801 tgggaggtct atataagcag agctctctgg ctaactagag aacccactgc ttactggctt
     4861 atcgaaatta atacgactca ctatagggag acccaagctg gctagccacc
//
