LOCUS       sequence-363965-        6739 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For the mammalian expression of the tardigrade protein 
            CAHS1_PARRC_101-189 attached to 2xFKBP. The FKBP can be used for 
            inducible dimerization and condensate formation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6739)
  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 6739)
  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_101-189_mCh-2xFKBP (pBS1136)
FEATURES             Location/Qualifiers
     source          1..6739
                     /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"
     protein_bind    816..834
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    837..855
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     865..889
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      998..1007
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2015..2335
                     /codon_start=1
                     /product="F36V mutant of FK506-binding protein FKBP12"
                     /label=DmrB
                     /note="binds synthetic ligands that do not bind wild-type 
                     FKBP"
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKVDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     CDS             2342..2662
                     /codon_start=1
                     /product="F36V mutant of FK506-binding protein FKBP12"
                     /label=DmrB
                     /note="binds synthetic ligands that do not bind wild-type 
                     FKBP"
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKVDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     CDS             2669..2695
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(2743..2760)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2749..2973
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3019..3447
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3106..3125)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3316..3337
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3456..3476)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3461..3790
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3641..3776
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3703..3722
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3838..3885
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3904..4278
                     /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    4408..4529
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4445..4464)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4499..4518
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4578..4594)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4578..4594)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4591..4613)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4602..4618
                     /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(4626..4632,4633..4650,4651..4656))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4671..4692
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4809..4826)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4980..5568)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5060..5079)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5739..6530,6531..6599))
                     /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     6362..6381
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6600..6704)
                     /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 agctctccct atcagtgata gagatctccc
      841 tatcagtgat agagatcgtc gacgagctcg tttagtgaac cgtcagatcg cctggagacg
      901 ccatccacgc tgttttgacc tccatagaag acaccgggac cgatccagcc tccggactct
      961 agcgtttaaa cttaagcttg gtaccgagct cggatccgcc accatggcca aaacgcgcga
     1021 acaggagatg ctgtctgcta actatcaaaa agaagttgag cgcaaaactg aagcctatcg
     1081 gaagcagcaa gaagtggagg cagacaaaat tcggaaagag ctggaaaagc agcatctgcg
     1141 tgacgtggag ttccgcaaag atattgtaga gatggcaatc gaaaaccaga aaaaaatgat
     1201 tgatgtcgaa agccgttatg caaaaaaaga tatggaccgc gaacgtgtga aagtccgtat
     1261 gatgctcgag cagtccggag gcggcggatc cggcggaatg gtgtctaaag gcgaggagga
     1321 taacatggct ataatcaagg aatttatgag gttcaaggtg catatggagg gatctgtgaa
     1381 cggtcacgag ttcgaaatcg agggcgaagg cgaggggcga ccctatgaag ggacacagac
     1441 ggccaaactg aaggtgacca agggtggccc cctgcctttc gcctgggaca tcctgtcacc
     1501 ccagtttatg tacggctcta aggcttacgt taagcaccct gccgatattc cggactactt
     1561 gaaactgagc ttccctgaag ggttcaagtg ggaacgggta atgaattttg aggatggcgg
     1621 tgtagtcaca gttacccagg acagctctct tcaggacgga gaatttatct ataaggttaa
     1681 acttcggggc actaacttcc catcagacgg ccccgtgatg cagaagaaaa ctatgggctg
     1741 ggaagccagc tcagaacgca tgtatcccga ggacggagca ctgaagggag aaatcaagca
     1801 gcgactcaag ctgaaggatg ggggacatta tgatgctgag gtcaaaacca cctataaggc
     1861 caagaaaccc gttcagcttc ctggggcata caacgtgaat atcaaactgg atattacatc
     1921 ccataacgaa gactatacca tcgtggaaca gtacgagcgg gccgagggca gacatagcac
     1981 agggggcatg gatgaattgt acaaggcgac tagaggcgtc caagtcgaaa ccattagtcc
     2041 cggcgatggc agaacatttc ctaaaagggg acaaacatgt gtcgtccatt atacaggcat
     2101 gttggaggac ggcaaaaagg tggacagtag tagagatcgc aataaacctt tcaaattcat
     2161 gttgggaaaa caagaagtca ttaggggatg ggaggagggc gtggctcaaa tgtccgtcgg
     2221 ccaacgcgct aagctcacca tcagccccga ctacgcatac ggcgctaccg gacatcccgg
     2281 aattattccc cctcacgcta ccttggtgtt tgacgtcgaa ctgttgaagc tcgagactag
     2341 aggagtgcag gtggagacta tctccccagg agacgggcgc accttcccca agcgcggcca
     2401 gacctgcgtg gtgcactaca ccgggatgct tgaagatgga aagaaagttg attcctcccg
     2461 ggacagaaac aagcccttta agtttatgct aggcaagcag gaggtgatcc gaggctggga
     2521 agaaggggtt gcccagatga gtgtgggtca gagagccaaa ctgactatat ctccagatta
     2581 tgcctatggt gccactgggc acccaggcat catcccacca catgccactc tcgtcttcga
     2641 tgtggagctt ctaaaactgg aaactagtta tccgtacgac gtaccagact acgcataagc
     2701 ggccgctcga gtctagaggg cccgtttaaa cccgctgatc agcctcgact gtgccttcta
     2761 gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca
     2821 ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc
     2881 attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata
     2941 gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg
     3001 gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     3061 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     3121 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     3181 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
     3241 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     3301 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     3361 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
     3421 tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg
     3481 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     3541 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     3601 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     3661 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     3721 ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     3781 cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc agcacgtgtt
     3841 gacaattaat catcggcata gtatatcggc atagtataat acgacaaggt gaggaactaa
     3901 accatggcca agttgaccag tgccgttccg gtgctcaccg cgcgcgacgt cgccggagcg
     3961 gtcgagttct ggaccgaccg gctcgggttc tcccgggact tcgtggagga cgacttcgcc
     4021 ggtgtggtcc gggacgacgt gaccctgttc atcagcgcgg tccaggacca ggtggtgccg
     4081 gacaacaccc tggcctgggt gtgggtgcgc ggcctggacg agctgtacgc cgagtggtcg
     4141 gaggtcgtgt ccacgaactt ccgggacgcc tccgggccgg ccatgaccga gatcggcgag
     4201 cagccgtggg ggcgggagtt cgccctgcgc gacccggccg gcaactgcgt gcacttcgtg
     4261 gccgaggagc aggactgaca cgtgctacga gatttcgatt ccaccgccgc cttctatgaa
     4321 aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat
     4381 ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa
     4441 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     4501 ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag
     4561 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     4621 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     4681 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     4741 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     4801 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     4861 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     4921 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     4981 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     5041 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     5101 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     5161 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     5221 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     5281 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     5341 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     5401 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     5461 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     5521 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     5581 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     5641 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     5701 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     5761 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     5821 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     5881 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     5941 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     6001 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     6061 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     6121 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     6181 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     6241 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     6301 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     6361 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     6421 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     6481 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     6541 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     6601 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     6661 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     6721 gtgccacctg acgtcgacg
//
