LOCUS       sequence-363956-        6517 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For the mammalian expression of the sleeping chironomid protein 
            pvLEA22mer_shuffle_3 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 6517)
  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 6517)
  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: pvLEA22mer_shuffle_3_mCh-2xFKBP (pBS1119)
FEATURES             Location/Qualifiers
     source          1..6517
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(45..64)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        236..615
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        616..819
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     770..790
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    821..839
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    842..860
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     870..894
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      1003..1012
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1819..2139
                     /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             2146..2466
                     /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             2473..2499
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(2547..2564)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2553..2777
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2823..3251
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2910..2929)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3120..3141
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3260..3280)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3425..3559
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3486..3505
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3621..3668
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3687..4061
                     /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    4191..4312
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4228..4247)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4282..4301
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4361..4377)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4361..4377)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4374..4396)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4385..4401
                     /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(4409..4415,4416..4433,4434..4439))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4454..4475
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4592..4609)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4763..5351)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4843..4862)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5522..6313,6314..6382))
                     /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     6145..6164
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6383..6487)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cgacggatcg ggagatctcc cgatccccta tggtgcactc tcagtacaat ctgctctgat
       61 gccgcatagt taagccagta tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc
      121 gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg acaattgcat gaagaatctg
      181 cttagggtta ggcgttttgc gctgcttcgc gatgtacggg ccagatatac gcgttgacat
      241 tgattattga ctagttatta atagtaatca attacggggt cattagttca tagcccatat
      301 atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac
      361 ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc
      421 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg
      481 tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat
      541 tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc
      601 atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt
      661 gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac
      721 caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc
      781 ggtaggcgtg tacggtggga ggtctatata agcagagctc tccctatcag tgatagagat
      841 ctccctatca gtgatagaga tcgtcgacga gctcgtttag tgaaccgtca gatcgcctgg
      901 agacgccatc cacgctgttt tgacctccat agaagacacc gggaccgatc cagcctccgg
      961 actctagcgt ttaaacttaa gcttggtacc gagctcggat ccgccaccat ggatggaacc
     1021 ggaaccaagg gagaggctga gaaggagaag aaggacaagg ccgccgaggc caagggctcc
     1081 ggaggcggcg gatccggcgg aatggtgtct aaaggcgagg aggataacat ggctataatc
     1141 aaggaattta tgaggttcaa ggtgcatatg gagggatctg tgaacggtca cgagttcgaa
     1201 atcgagggcg aaggcgaggg gcgaccctat gaagggacac agacggccaa actgaaggtg
     1261 accaagggtg gccccctgcc tttcgcctgg gacatcctgt caccccagtt tatgtacggc
     1321 tctaaggctt acgttaagca ccctgccgat attccggact acttgaaact gagcttccct
     1381 gaagggttca agtgggaacg ggtaatgaat tttgaggatg gcggtgtagt cacagttacc
     1441 caggacagct ctcttcagga cggagaattt atctataagg ttaaacttcg gggcactaac
     1501 ttcccatcag acggccccgt gatgcagaag aaaactatgg gctgggaagc cagctcagaa
     1561 cgcatgtatc ccgaggacgg agcactgaag ggagaaatca agcagcgact caagctgaag
     1621 gatgggggac attatgatgc tgaggtcaaa accacctata aggccaagaa acccgttcag
     1681 cttcctgggg catacaacgt gaatatcaaa ctggatatta catcccataa cgaagactat
     1741 accatcgtgg aacagtacga gcgggccgag ggcagacata gcacaggggg catggatgaa
     1801 ttgtacaagg cgactagagg cgtccaagtc gaaaccatta gtcccggcga tggcagaaca
     1861 tttcctaaaa ggggacaaac atgtgtcgtc cattatacag gcatgttgga ggacggcaaa
     1921 aaggtggaca gtagtagaga tcgcaataaa cctttcaaat tcatgttggg aaaacaagaa
     1981 gtcattaggg gatgggagga gggcgtggct caaatgtccg tcggccaacg cgctaagctc
     2041 accatcagcc ccgactacgc atacggcgct accggacatc ccggaattat tccccctcac
     2101 gctaccttgg tgtttgacgt cgaactgttg aagctcgaga ctagaggagt gcaggtggag
     2161 actatctccc caggagacgg gcgcaccttc cccaagcgcg gccagacctg cgtggtgcac
     2221 tacaccggga tgcttgaaga tggaaagaaa gttgattcct cccgggacag aaacaagccc
     2281 tttaagttta tgctaggcaa gcaggaggtg atccgaggct gggaagaagg ggttgcccag
     2341 atgagtgtgg gtcagagagc caaactgact atatctccag attatgccta tggtgccact
     2401 gggcacccag gcatcatccc accacatgcc actctcgtct tcgatgtgga gcttctaaaa
     2461 ctggaaacta gttatccgta cgacgtacca gactacgcat aagcggccgc tcgagtctag
     2521 agggcccgtt taaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt
     2581 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc
     2641 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg
     2701 tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga
     2761 tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc
     2821 ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     2881 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     2941 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     3001 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     3061 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     3121 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     3181 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     3241 taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     3301 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     3361 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     3421 atagtcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     3481 ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga
     3541 ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt
     3601 ttcggatctg atcagcacgt gttgacaatt aatcatcggc atagtatatc ggcatagtat
     3661 aatacgacaa ggtgaggaac taaaccatgg ccaagttgac cagtgccgtt ccggtgctca
     3721 ccgcgcgcga cgtcgccgga gcggtcgagt tctggaccga ccggctcggg ttctcccggg
     3781 acttcgtgga ggacgacttc gccggtgtgg tccgggacga cgtgaccctg ttcatcagcg
     3841 cggtccagga ccaggtggtg ccggacaaca ccctggcctg ggtgtgggtg cgcggcctgg
     3901 acgagctgta cgccgagtgg tcggaggtcg tgtccacgaa cttccgggac gcctccgggc
     3961 cggccatgac cgagatcggc gagcagccgt gggggcggga gttcgccctg cgcgacccgg
     4021 ccggcaactg cgtgcacttc gtggccgagg agcaggactg acacgtgcta cgagatttcg
     4081 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
     4141 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta
     4201 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     4261 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
     4321 gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt
     4381 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     4441 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     4501 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     4561 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     4621 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     4681 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     4741 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     4801 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     4861 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     4921 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     4981 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     5041 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     5101 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     5161 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     5221 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     5281 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     5341 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     5401 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     5461 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     5521 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     5581 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     5641 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     5701 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     5761 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     5821 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     5881 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     5941 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     6001 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     6061 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     6121 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     6181 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     6241 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     6301 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     6361 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     6421 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     6481 ttccgcgcac atttccccga aaagtgccac ctgacgt
//
