LOCUS       sequence-363959-        6622 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For the mammalian expression of the sleeping chironomid protein 
            PvLEA4_repeats_k5_1 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_k5_1_mCh-2xFKBP (pBS1122)
FEATURES             Location/Qualifiers
     source          1..6622
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(48..67)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        239..618
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        619..822
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     773..793
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    824..842
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    845..863
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     873..897
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             1906..2226
                     /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             2233..2553
                     /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             2560..2586
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(2634..2651)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2640..2864
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2910..3338
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2997..3016)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3207..3228
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3347..3367)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3352..3681
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3532..3667
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3594..3613
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3729..3776
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3795..4169
                     /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    4299..4420
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4336..4355)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4390..4409
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4469..4485)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4469..4485)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4482..4504)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4493..4509
                     /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(4517..4523,4524..4541,4542..4547))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4562..4583
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4700..4717)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4871..5459)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4951..4970)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5630..6421,6422..6490))
                     /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     6253..6272
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6491..6595)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cgtcgacgga tcgggagatc tcccgatccc ctatggtgca ctctcagtac aatctgctct
       61 gatgccgcat agttaagcca gtatctgctc cctgcttgtg tgttggaggt cgctgagtag
      121 tgcgcgagca aaatttaagc tacaacaagg caaggcttga ccgacaattg catgaagaat
      181 ctgcttaggg ttaggcgttt tgcgctgctt cgcgatgtac gggccagata tacgcgttga
      241 cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca
      301 tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac
      361 gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact
      421 ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa
      481 gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg
      541 cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta
      601 gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg
      661 tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg
      721 caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg
      781 ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctctccctat cagtgataga
      841 gatctcccta tcagtgatag agatcgtcga cgagctcgtt tagtgaaccg tcagatcgcc
      901 tggagacgcc atccacgctg ttttgacctc catagaagac accgggaccg atccagcctc
      961 cggactctag cgtttaaact taagcttggt accgagctcg gatccgccac catgttccag
     1021 accgccgccg acaaggccgc cgccgccaag gacaccgttg ttgacacctt gtgcgacgcc
     1081 aaggacaccg ccaaggagaa ggtggtgggc gccaaggacg ccggagccga ctacgctggc
     1141 gagaagaagg agcagatgag ggactccgga ggcggcggat ccggcggaat ggtgtctaaa
     1201 ggcgaggagg ataacatggc tataatcaag gaatttatga ggttcaaggt gcatatggag
     1261 ggatctgtga acggtcacga gttcgaaatc gagggcgaag gcgaggggcg accctatgaa
     1321 gggacacaga cggccaaact gaaggtgacc aagggtggcc ccctgccttt cgcctgggac
     1381 atcctgtcac cccagtttat gtacggctct aaggcttacg ttaagcaccc tgccgatatt
     1441 ccggactact tgaaactgag cttccctgaa gggttcaagt gggaacgggt aatgaatttt
     1501 gaggatggcg gtgtagtcac agttacccag gacagctctc ttcaggacgg agaatttatc
     1561 tataaggtta aacttcgggg cactaacttc ccatcagacg gccccgtgat gcagaagaaa
     1621 actatgggct gggaagccag ctcagaacgc atgtatcccg aggacggagc actgaaggga
     1681 gaaatcaagc agcgactcaa gctgaaggat gggggacatt atgatgctga ggtcaaaacc
     1741 acctataagg ccaagaaacc cgttcagctt cctggggcat acaacgtgaa tatcaaactg
     1801 gatattacat cccataacga agactatacc atcgtggaac agtacgagcg ggccgagggc
     1861 agacatagca cagggggcat ggatgaattg tacaaggcga ctagaggcgt ccaagtcgaa
     1921 accattagtc ccggcgatgg cagaacattt cctaaaaggg gacaaacatg tgtcgtccat
     1981 tatacaggca tgttggagga cggcaaaaag gtggacagta gtagagatcg caataaacct
     2041 ttcaaattca tgttgggaaa acaagaagtc attaggggat gggaggaggg cgtggctcaa
     2101 atgtccgtcg gccaacgcgc taagctcacc atcagccccg actacgcata cggcgctacc
     2161 ggacatcccg gaattattcc ccctcacgct accttggtgt ttgacgtcga actgttgaag
     2221 ctcgagacta gaggagtgca ggtggagact atctccccag gagacgggcg caccttcccc
     2281 aagcgcggcc agacctgcgt ggtgcactac accgggatgc ttgaagatgg aaagaaagtt
     2341 gattcctccc gggacagaaa caagcccttt aagtttatgc taggcaagca ggaggtgatc
     2401 cgaggctggg aagaaggggt tgcccagatg agtgtgggtc agagagccaa actgactata
     2461 tctccagatt atgcctatgg tgccactggg cacccaggca tcatcccacc acatgccact
     2521 ctcgtcttcg atgtggagct tctaaaactg gaaactagtt atccgtacga cgtaccagac
     2581 tacgcataag cggccgctcg agtctagagg gcccgtttaa acccgctgat cagcctcgac
     2641 tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct
     2701 ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct
     2761 gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg
     2821 ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag
     2881 aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc
     2941 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     3001 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     3061 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     3121 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     3181 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     3241 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     3301 aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag
     3361 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     3421 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     3481 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     3541 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     3601 gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt
     3661 ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat
     3721 cagcacgtgt tgacaattaa tcatcggcat agtatatcgg catagtataa tacgacaagg
     3781 tgaggaacta aaccatggcc aagttgacca gtgccgttcc ggtgctcacc gcgcgcgacg
     3841 tcgccggagc ggtcgagttc tggaccgacc ggctcgggtt ctcccgggac ttcgtggagg
     3901 acgacttcgc cggtgtggtc cgggacgacg tgaccctgtt catcagcgcg gtccaggacc
     3961 aggtggtgcc ggacaacacc ctggcctggg tgtgggtgcg cggcctggac gagctgtacg
     4021 ccgagtggtc ggaggtcgtg tccacgaact tccgggacgc ctccgggccg gccatgaccg
     4081 agatcggcga gcagccgtgg gggcgggagt tcgccctgcg cgacccggcc ggcaactgcg
     4141 tgcacttcgt ggccgaggag caggactgac acgtgctacg agatttcgat tccaccgccg
     4201 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
     4261 agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata
     4321 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     4381 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga
     4441 cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     4501 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     4561 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     4621 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     4681 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     4741 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     4801 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     4861 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     4921 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     4981 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     5041 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     5101 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     5161 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     5221 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     5281 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     5341 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     5401 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     5461 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     5521 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     5581 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     5641 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     5701 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     5761 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     5821 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     5881 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     5941 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     6001 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     6061 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     6121 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     6181 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     6241 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     6301 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     6361 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     6421 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     6481 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     6541 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     6601 ttccccgaaa agtgccacct ga
//
