LOCUS       sequence-363952-        6859 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For the mammalian expression of the riverbank grape plant protein 
            DHN_VrDHN1a 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 6859)
  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 6859)
  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: DHN_VrDHN1a_mCh-2xFKBP (pBS1117)
FEATURES             Location/Qualifiers
     source          1..6859
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(50..69)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        241..620
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        621..824
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     775..795
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    826..844
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    847..865
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     875..899
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      1008..1017
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2145..2465
                     /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             2472..2792
                     /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             2799..2825
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(2873..2890)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2879..3103
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3149..3577
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3236..3255)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3446..3467
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3586..3606)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3591..3920
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3771..3906
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3833..3852
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3968..4015
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4034..4408
                     /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    4538..4659
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4575..4594)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4629..4648
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4708..4724)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4708..4724)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4721..4743)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4732..4748
                     /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(4756..4762,4763..4780,4781..4786))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4801..4822
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4939..4956)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5110..5698)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5190..5209)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5869..6660,6661..6729))
                     /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     6492..6511
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6730..6834)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacgtcgacg gatcgggaga tctcccgatc ccctatggtg cactctcagt acaatctgct
       61 ctgatgccgc atagttaagc cagtatctgc tccctgcttg tgtgttggag gtcgctgagt
      121 agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga
      181 atctgcttag ggttaggcgt tttgcgctgc ttcgcgatgt acgggccaga tatacgcgtt
      241 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
      301 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      361 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      421 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      481 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      541 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      601 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      661 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      721 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      781 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctccct atcagtgata
      841 gagatctccc tatcagtgat agagatcgtc gacgagctcg tttagtgaac cgtcagatcg
      901 cctggagacg ccatccacgc tgttttgacc tccatagaag acaccgggac cgatccagcc
      961 tccggactct agcgtttaaa cttaagcttg gtaccgagct cggatccgcc accatggcct
     1021 accagcagga cccctgcgcc aaccccacca ggcagaccgg aaagaccgga ggacagaccg
     1081 accagtacgg caaccccgtg caccagaccg aggccctggg agcctacgga gctggaaccg
     1141 gaaccggcat gcacggagga gagcaccagc agcagcccca ccagcagcct ggagtgctga
     1201 acaggagcgg cagcagcagc agcgaggacg acggccaggg aggaaggagg aagaagggca
     1261 tgaaggagaa gatcaaggag aggatccccg gcatgggcag gaaggacgag cagaagcaga
     1321 ccagcgccac cagcaccccc ggacagggcc aacagcagaa gggcatgatg gagaagatca
     1381 aggagaagct gcccggcgcc cactccggag gcggcggatc cggcggaatg gtgtctaaag
     1441 gcgaggagga taacatggct ataatcaagg aatttatgag gttcaaggtg catatggagg
     1501 gatctgtgaa cggtcacgag ttcgaaatcg agggcgaagg cgaggggcga ccctatgaag
     1561 ggacacagac ggccaaactg aaggtgacca agggtggccc cctgcctttc gcctgggaca
     1621 tcctgtcacc ccagtttatg tacggctcta aggcttacgt taagcaccct gccgatattc
     1681 cggactactt gaaactgagc ttccctgaag ggttcaagtg ggaacgggta atgaattttg
     1741 aggatggcgg tgtagtcaca gttacccagg acagctctct tcaggacgga gaatttatct
     1801 ataaggttaa acttcggggc actaacttcc catcagacgg ccccgtgatg cagaagaaaa
     1861 ctatgggctg ggaagccagc tcagaacgca tgtatcccga ggacggagca ctgaagggag
     1921 aaatcaagca gcgactcaag ctgaaggatg ggggacatta tgatgctgag gtcaaaacca
     1981 cctataaggc caagaaaccc gttcagcttc ctggggcata caacgtgaat atcaaactgg
     2041 atattacatc ccataacgaa gactatacca tcgtggaaca gtacgagcgg gccgagggca
     2101 gacatagcac agggggcatg gatgaattgt acaaggcgac tagaggcgtc caagtcgaaa
     2161 ccattagtcc cggcgatggc agaacatttc ctaaaagggg acaaacatgt gtcgtccatt
     2221 atacaggcat gttggaggac ggcaaaaagg tggacagtag tagagatcgc aataaacctt
     2281 tcaaattcat gttgggaaaa caagaagtca ttaggggatg ggaggagggc gtggctcaaa
     2341 tgtccgtcgg ccaacgcgct aagctcacca tcagccccga ctacgcatac ggcgctaccg
     2401 gacatcccgg aattattccc cctcacgcta ccttggtgtt tgacgtcgaa ctgttgaagc
     2461 tcgagactag aggagtgcag gtggagacta tctccccagg agacgggcgc accttcccca
     2521 agcgcggcca gacctgcgtg gtgcactaca ccgggatgct tgaagatgga aagaaagttg
     2581 attcctcccg ggacagaaac aagcccttta agtttatgct aggcaagcag gaggtgatcc
     2641 gaggctggga agaaggggtt gcccagatga gtgtgggtca gagagccaaa ctgactatat
     2701 ctccagatta tgcctatggt gccactgggc acccaggcat catcccacca catgccactc
     2761 tcgtcttcga tgtggagctt ctaaaactgg aaactagtta tccgtacgac gtaccagact
     2821 acgcataagc ggccgctcga gtctagaggg cccgtttaaa cccgctgatc agcctcgact
     2881 gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg
     2941 gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg
     3001 agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg
     3061 gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga
     3121 accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg
     3181 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     3241 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     3301 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     3361 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     3421 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     3481 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     3541 aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt
     3601 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     3661 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     3721 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     3781 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     3841 cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg
     3901 gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc
     3961 agcacgtgtt gacaattaat catcggcata gtatatcggc atagtataat acgacaaggt
     4021 gaggaactaa accatggcca agttgaccag tgccgttccg gtgctcaccg cgcgcgacgt
     4081 cgccggagcg gtcgagttct ggaccgaccg gctcgggttc tcccgggact tcgtggagga
     4141 cgacttcgcc ggtgtggtcc gggacgacgt gaccctgttc atcagcgcgg tccaggacca
     4201 ggtggtgccg gacaacaccc tggcctgggt gtgggtgcgc ggcctggacg agctgtacgc
     4261 cgagtggtcg gaggtcgtgt ccacgaactt ccgggacgcc tccgggccgg ccatgaccga
     4321 gatcggcgag cagccgtggg ggcgggagtt cgccctgcgc gacccggccg gcaactgcgt
     4381 gcacttcgtg gccgaggagc aggactgaca cgtgctacga gatttcgatt ccaccgccgc
     4441 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     4501 gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa
     4561 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     4621 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac
     4681 ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     4741 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     4801 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     4861 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     4921 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     4981 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     5041 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     5101 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     5161 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     5221 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     5281 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     5341 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     5401 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     5461 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     5521 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     5581 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     5641 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     5701 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     5761 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     5821 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     5881 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     5941 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     6001 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     6061 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     6121 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     6181 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     6241 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     6301 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     6361 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     6421 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     6481 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     6541 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     6601 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     6661 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     6721 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     6781 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     6841 tccccgaaaa gtgccacct
//
