LOCUS       sequence-363953-        6622 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For the mammalian expression of the sleeping chironomid protein 
            PvLEA4_repeats_k3_26 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 6622)
  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 6622)
  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: PvLEA4_repeats_k3_26_mCh-2xFKBP (pBS1118)
FEATURES             Location/Qualifiers
     source          1..6622
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(58..77)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        249..628
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        629..832
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     783..803
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    834..852
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    855..873
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     883..907
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             1916..2236
                     /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             2243..2563
                     /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             2570..2596
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(2644..2661)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2650..2874
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2920..3348
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3007..3026)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3217..3238
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3357..3377)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3362..3691
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3542..3677
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3604..3623
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3739..3786
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3805..4179
                     /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    4309..4430
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4346..4365)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4400..4419
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4479..4495)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4479..4495)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4492..4514)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4503..4519
                     /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(4527..4533,4534..4551,4552..4557))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4572..4593
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4710..4727)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4881..5469)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4961..4980)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5640..6431,6432..6500))
                     /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     6263..6282
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6501..6605)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tgccacctga cgtcgacgga tcgggagatc tcccgatccc ctatggtgca ctctcagtac
       61 aatctgctct gatgccgcat agttaagcca gtatctgctc cctgcttgtg tgttggaggt
      121 cgctgagtag tgcgcgagca aaatttaagc tacaacaagg caaggcttga ccgacaattg
      181 catgaagaat ctgcttaggg ttaggcgttt tgcgctgctt cgcgatgtac gggccagata
      241 tacgcgttga cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt
      301 tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg
      361 accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc
      421 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc
      481 agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg
      541 gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat
      601 ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg
      661 tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag
      721 tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt
      781 gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctctccctat
      841 cagtgataga gatctcccta tcagtgatag agatcgtcga cgagctcgtt tagtgaaccg
      901 tcagatcgcc tggagacgcc atccacgctg ttttgacctc catagaagac accgggaccg
      961 atccagcctc cggactctag cgtttaaact taagcttggt accgagctcg gatccgccac
     1021 catgttccag accgccgccg acaagctggc cgccgccaag gacaccgtgg ttgacacctt
     1081 gggcagcgcc aaggacaccg tggtggagaa ggtggtgggc gccaaggacg ccaccgccga
     1141 gtacctggga gagaagaagg agcagatgag ggactccgga ggcggcggat ccggcggaat
     1201 ggtgtctaaa ggcgaggagg ataacatggc tataatcaag gaatttatga ggttcaaggt
     1261 gcatatggag ggatctgtga acggtcacga gttcgaaatc gagggcgaag gcgaggggcg
     1321 accctatgaa gggacacaga cggccaaact gaaggtgacc aagggtggcc ccctgccttt
     1381 cgcctgggac atcctgtcac cccagtttat gtacggctct aaggcttacg ttaagcaccc
     1441 tgccgatatt ccggactact tgaaactgag cttccctgaa gggttcaagt gggaacgggt
     1501 aatgaatttt gaggatggcg gtgtagtcac agttacccag gacagctctc ttcaggacgg
     1561 agaatttatc tataaggtta aacttcgggg cactaacttc ccatcagacg gccccgtgat
     1621 gcagaagaaa actatgggct gggaagccag ctcagaacgc atgtatcccg aggacggagc
     1681 actgaaggga gaaatcaagc agcgactcaa gctgaaggat gggggacatt atgatgctga
     1741 ggtcaaaacc acctataagg ccaagaaacc cgttcagctt cctggggcat acaacgtgaa
     1801 tatcaaactg gatattacat cccataacga agactatacc atcgtggaac agtacgagcg
     1861 ggccgagggc agacatagca cagggggcat ggatgaattg tacaaggcga ctagaggcgt
     1921 ccaagtcgaa accattagtc ccggcgatgg cagaacattt cctaaaaggg gacaaacatg
     1981 tgtcgtccat tatacaggca tgttggagga cggcaaaaag gtggacagta gtagagatcg
     2041 caataaacct ttcaaattca tgttgggaaa acaagaagtc attaggggat gggaggaggg
     2101 cgtggctcaa atgtccgtcg gccaacgcgc taagctcacc atcagccccg actacgcata
     2161 cggcgctacc ggacatcccg gaattattcc ccctcacgct accttggtgt ttgacgtcga
     2221 actgttgaag ctcgagacta gaggagtgca ggtggagact atctccccag gagacgggcg
     2281 caccttcccc aagcgcggcc agacctgcgt ggtgcactac accgggatgc ttgaagatgg
     2341 aaagaaagtt gattcctccc gggacagaaa caagcccttt aagtttatgc taggcaagca
     2401 ggaggtgatc cgaggctggg aagaaggggt tgcccagatg agtgtgggtc agagagccaa
     2461 actgactata tctccagatt atgcctatgg tgccactggg cacccaggca tcatcccacc
     2521 acatgccact ctcgtcttcg atgtggagct tctaaaactg gaaactagtt atccgtacga
     2581 cgtaccagac tacgcataag cggccgctcg agtctagagg gcccgtttaa acccgctgat
     2641 cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt
     2701 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat
     2761 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg
     2821 gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg
     2881 aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat
     2941 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     3001 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     3061 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     3121 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     3181 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     3241 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     3301 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa
     3361 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     3421 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     3481 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     3541 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     3601 ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg
     3661 aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt
     3721 cggatctgat cagcacgtgt tgacaattaa tcatcggcat agtatatcgg catagtataa
     3781 tacgacaagg tgaggaacta aaccatggcc aagttgacca gtgccgttcc ggtgctcacc
     3841 gcgcgcgacg tcgccggagc ggtcgagttc tggaccgacc ggctcgggtt ctcccgggac
     3901 ttcgtggagg acgacttcgc cggtgtggtc cgggacgacg tgaccctgtt catcagcgcg
     3961 gtccaggacc aggtggtgcc ggacaacacc ctggcctggg tgtgggtgcg cggcctggac
     4021 gagctgtacg ccgagtggtc ggaggtcgtg tccacgaact tccgggacgc ctccgggccg
     4081 gccatgaccg agatcggcga gcagccgtgg gggcgggagt tcgccctgcg cgacccggcc
     4141 ggcaactgcg tgcacttcgt ggccgaggag caggactgac acgtgctacg agatttcgat
     4201 tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg
     4261 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt
     4321 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     4381 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt
     4441 ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga
     4501 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     4561 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     4621 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     4681 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     4741 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     4801 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     4861 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     4921 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     4981 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     5041 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     5101 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     5161 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     5221 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     5281 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     5341 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     5401 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     5461 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     5521 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     5581 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     5641 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     5701 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     5761 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     5821 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     5881 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     5941 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     6001 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     6061 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     6121 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     6181 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     6241 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     6301 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     6361 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     6421 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     6481 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     6541 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     6601 ccgcgcacat ttccccgaaa ag
//
