LOCUS       sequence-363940-        7141 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For the mammalian expression of the tardigrade protein CAHS3hypdu 
            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 7141)
  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 7141)
  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: CAHS3hypdu-mCh-2xFKBP (pBS1110)
FEATURES             Location/Qualifiers
     source          1..7141
                     /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"
     protein_bind    831..849
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    852..870
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     880..904
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             2432..2752
                     /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             2759..3079
                     /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             3086..3112
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(3160..3177)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3166..3390
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3436..3864
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3523..3542)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3733..3754
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3873..3893)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3878..4207
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4058..4193
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4120..4139
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4255..4302
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4321..4695
                     /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    4825..4946
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4862..4881)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4916..4935
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4995..5011)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4995..5011)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5008..5030)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5019..5035
                     /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(5043..5049,5050..5067,5068..5073))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5088..5109
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5226..5243)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5397..5985)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5477..5496)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6156..6947,6948..7016))
                     /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     6779..6798
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7017..7121)
                     /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 tccctatcag
      841 tgatagagat ctccctatca gtgatagaga tcgtcgacga gctcgtttag tgaaccgtca
      901 gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagacacc gggaccgatc
      961 cagcctccgg actctagcgt ttaaacttaa gcttggtacc gagctcggat ccgccaccat
     1021 gagtaattac caacaagaga gctcatacca atattcagat cgctccaaca acgggcaaca
     1081 acaagagcaa caggagaaga aagaggtaga acacagtagc tacacacata ccgacgtgaa
     1141 agtcaacatg ccaaatctga tcgcgccatt cattagtagc agcgccggcc tcgcccaaga
     1201 actcgttgga gaagggttcc aggcttctgt ttctaggatc accggagcat ccggtgaact
     1261 gaccgtcatt gacacagaag cagaaaccga agaggcacga agagatatgg aagccaaggc
     1321 ccgcgaacag gaactgctgt cacgccaatt cgagaaagag cttgagcgta aaacagaagc
     1381 ctacagaaag caacaagaag ttgaaaccga gaaaatacgg aaagaactcg agaaacaaca
     1441 cttgcgggac gtagaattcc gaaaagagtt gatggaacaa actattgaaa accagaaaag
     1501 gcaaattgat ctggaagctc gctacgcaaa gaaggaactg gaacgggagc ggaacaaagt
     1561 gaagcgggtt ttggagcgga gcaagtttca tacggatata caggttaata tggaagcagc
     1621 cgcaggttct acgcactccg gatcatcatc cgtagccgtt agtgaatctg agaaatttca
     1681 aaccaataac tccggaggcg gcggatccgg cggaatggtg tctaaaggcg aggaggataa
     1741 catggctata atcaaggaat ttatgaggtt caaggtgcat atggagggat ctgtgaacgg
     1801 tcacgagttc gaaatcgagg gcgaaggcga ggggcgaccc tatgaaggga cacagacggc
     1861 caaactgaag gtgaccaagg gtggccccct gcctttcgcc tgggacatcc tgtcacccca
     1921 gtttatgtac ggctctaagg cttacgttaa gcaccctgcc gatattccgg actacttgaa
     1981 actgagcttc cctgaagggt tcaagtggga acgggtaatg aattttgagg atggcggtgt
     2041 agtcacagtt acccaggaca gctctcttca ggacggagaa tttatctata aggttaaact
     2101 tcggggcact aacttcccat cagacggccc cgtgatgcag aagaaaacta tgggctggga
     2161 agccagctca gaacgcatgt atcccgagga cggagcactg aagggagaaa tcaagcagcg
     2221 actcaagctg aaggatgggg gacattatga tgctgaggtc aaaaccacct ataaggccaa
     2281 gaaacccgtt cagcttcctg gggcatacaa cgtgaatatc aaactggata ttacatccca
     2341 taacgaagac tataccatcg tggaacagta cgagcgggcc gagggcagac atagcacagg
     2401 gggcatggat gaattgtaca aggcgactag aggcgtccaa gtcgaaacca ttagtcccgg
     2461 cgatggcaga acatttccta aaaggggaca aacatgtgtc gtccattata caggcatgtt
     2521 ggaggacggc aaaaaggtgg acagtagtag agatcgcaat aaacctttca aattcatgtt
     2581 gggaaaacaa gaagtcatta ggggatggga ggagggcgtg gctcaaatgt ccgtcggcca
     2641 acgcgctaag ctcaccatca gccccgacta cgcatacggc gctaccggac atcccggaat
     2701 tattccccct cacgctacct tggtgtttga cgtcgaactg ttgaagctcg agactagagg
     2761 agtgcaggtg gagactatct ccccaggaga cgggcgcacc ttccccaagc gcggccagac
     2821 ctgcgtggtg cactacaccg ggatgcttga agatggaaag aaagttgatt cctcccggga
     2881 cagaaacaag ccctttaagt ttatgctagg caagcaggag gtgatccgag gctgggaaga
     2941 aggggttgcc cagatgagtg tgggtcagag agccaaactg actatatctc cagattatgc
     3001 ctatggtgcc actgggcacc caggcatcat cccaccacat gccactctcg tcttcgatgt
     3061 ggagcttcta aaactggaaa ctagttatcc gtacgacgta ccagactacg cataagcggc
     3121 cgctcgagtc tagagggccc gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt
     3181 gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc
     3241 ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt
     3301 ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca
     3361 ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct
     3421 ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     3481 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     3541 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     3601 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     3661 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     3721 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
     3781 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     3841 tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa
     3901 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     3961 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     4021 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     4081 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     4141 cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     4201 ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcagc acgtgttgac
     4261 aattaatcat cggcatagta tatcggcata gtataatacg acaaggtgag gaactaaacc
     4321 atggccaagt tgaccagtgc cgttccggtg ctcaccgcgc gcgacgtcgc cggagcggtc
     4381 gagttctgga ccgaccggct cgggttctcc cgggacttcg tggaggacga cttcgccggt
     4441 gtggtccggg acgacgtgac cctgttcatc agcgcggtcc aggaccaggt ggtgccggac
     4501 aacaccctgg cctgggtgtg ggtgcgcggc ctggacgagc tgtacgccga gtggtcggag
     4561 gtcgtgtcca cgaacttccg ggacgcctcc gggccggcca tgaccgagat cggcgagcag
     4621 ccgtgggggc gggagttcgc cctgcgcgac ccggccggca actgcgtgca cttcgtggcc
     4681 gaggagcagg actgacacgt gctacgagat ttcgattcca ccgccgcctt ctatgaaagg
     4741 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc
     4801 atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa
     4861 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     4921 ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc
     4981 ttggcgtaat catagtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     5041 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     5101 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     5161 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     5221 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     5281 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     5341 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     5401 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     5461 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     5521 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     5581 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     5641 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     5701 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     5761 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     5821 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     5881 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     5941 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     6001 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     6061 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     6121 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     6181 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     6241 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     6301 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     6361 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     6421 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     6481 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     6541 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     6601 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     6661 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     6721 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     6781 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     6841 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     6901 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     6961 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     7021 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     7081 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     7141 c
//
