LOCUS       sequence-363896-        6698 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For the mammalian expression of the synthetic protein 
            iLID::eGFP::FTH1 attached to SspB for light inducible binding to 
            iLID scaffolds. Systems like this can be used to induce condensate 
            formation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6698)
  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 6698)
  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: LLPS1-iLID::EGFP::FTH1 (pBS1042)
FEATURES             Location/Qualifiers
     source          1..6698
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(62..81)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        253..632
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        633..836
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     787..807
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     881..900
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        881..899
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             926..1363
                     /codon_start=1
                     /product="improved light-inducible dimer when used with 
                     SspB nano or SspB micro (Guntas et al., 2015)"
                     /label=iLID
                     /translation="LATTLERIEKNFVITDPRLPDNPIIFASDSFLQLTEYSREEILGR
                     NCRFLQGPETDRATVRKIRDAIDNQTEVTVQLINYTKSGKKFWNVFHLQPMRDYKGDVQ
                     YFIGVQLDGTERLHGAAEREAVCLIKKTAFQIAEAANDENYF"
     CDS             1394..2107
                     /codon_start=1
                     /product="S65T variant of Aequorea victoria green 
                     fluorescent protein (Heim et al., 1995)"
                     /label=GFP (S65T)
                     /note="excitable with blue light"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(1424..1452)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2047..2067
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(2724..2741)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2730..2954
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3000..3428
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3087..3106)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3297..3318
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3437..3457)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3442..3771
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3622..3757
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3684..3703
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3819..3866
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3885..4259
                     /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    4389..4510
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4426..4445)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4480..4499
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4559..4575)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4559..4575)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4572..4594)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4583..4599
                     /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(4607..4613,4614..4631,4632..4637))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(4790..4807)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4961..5549)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5041..5060)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5720..6511,6512..6580))
                     /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     6343..6362
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6581..6685)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 aaagtgccac ctgacgtcga cggatcggga gatctcccga tcccctatgg tgcactctca
       61 gtacaatctg ctctgatgcc gcatagttaa gccagtatct gctccctgct tgtgtgttgg
      121 aggtcgctga gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc ttgaccgaca
      181 attgcatgaa gaatctgctt agggttaggc gttttgcgct gcttcgcgat gtacgggcca
      241 gatatacgcg ttgacattga ttattgacta gttattaata gtaatcaatt acggggtcat
      301 tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg
      361 gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa
      421 cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact
      481 tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta
      541 aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt
      601 acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag tacatcaatg
      661 ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg
      721 ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc
      781 cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctctct
      841 ggctaactag agaacccact gcttactggc ttatcgaaat taatacgact cactataggg
      901 agacccaagc tggctagcca ccatgctggc aaccacactg gaacggatcg agaaaaattt
      961 cgtgattact gatccgagac tgcctgacaa cccaatcatt tttgcgagcg attccttcct
     1021 gcagctgaca gaatattctc gggaagagat cctggggcgc aattgccgtt ttctgcaggg
     1081 acccgagaca gaccgtgcca ctgttcggaa aatcagagat gctattgaca accagactga
     1141 agtgaccgtt cagctgatca attataccaa gagcggcaag aagttctgga acgtgttcca
     1201 cctgcagccg atgcgcgatt ataagggcga cgtccagtac ttcattggcg tgcagctgga
     1261 tggcaccgaa cgtcttcatg gcgccgctga gcgtgaggcg gtctgcctga tcaaaaagac
     1321 agcctttcag attgctgagg cagcgaacga cgaaaattac tttggcggct ccggctctgg
     1381 atccggagct accatgagta aaggagaaga acttttcact ggagttgtcc caattcttgt
     1441 tgaattagat ggtgatgtta atgggcacaa attttctgtc agtggagagg gtgaaggtga
     1501 tgcaacatac ggaaaactta cccttaaatt tatttgcact actggaaaac tacctgttcc
     1561 atggccaaca cttgtcacta ccctgactta tggtgttcaa tgcttttcaa gatacccaga
     1621 tcatatgaaa cggcatgact ttttcaagag tgccatgccc gaaggttatg tacaggaaag
     1681 aactatattt ttcaaagatg acgggaacta caagacacgt gctgaagtca agtttgaagg
     1741 tgataccctt gttaatagaa tcgagttaaa aggtattgat tttaaagaag atggaaacat
     1801 tcttggacac aaattggaat acaactataa ctcacacaat gtatacatca tggcagacaa
     1861 acaaaagaat ggaatcaaag ttaacttcaa aattagacac aacattgaag atggaagcgt
     1921 tcaactagca gaccattatc aacaaaatac tccaattggc gatggccctg tccttttacc
     1981 agacaaccat tacctgtcca cacaatctaa gctttcgaaa gatcccaacg aaaagagaga
     2041 ccacatggtc cttcttgagt ttgtaacagc tgctgggatt acacatggca tggatgaact
     2101 atacaaaggt ggatctggag gttcaggtgg aagtgcggcc ggtaccggca gcggaggaat
     2161 gacgaccgcg tccacctcgc aggtgcgcca gaactaccac caggactcag aggccgccat
     2221 caaccgccag atcaacctgg agctctacgc ctcctacgtt tacctgtcca tgtcttacta
     2281 ctttgaccgc gatgatgtgg ctttgaagaa ctttgccaaa tactttcttc accaatctca
     2341 tgaggagagg gaacatgctg agaaactgat gaagctgcag aaccaacgag gtggccgaat
     2401 cttccttcag gatatcaaga aaccagactg tgatgactgg gagagcgggc tgaatgcaat
     2461 ggagtgtgca ttacatttgg aaaaaaatgt gaatcagtca ctactggaac tgcacaaact
     2521 ggccactgac aaaaatgacc cccatttgtg tgacttcatt gagacacatt acctgaatga
     2581 gcaggtgaaa gccatcaaag aattgggtga ccacgtgacc aacttgcgca agatgggagc
     2641 gcccgaatct ggcttggcgg aatatctctt tgacaagcac accctgggag acagtgataa
     2701 tgaaagctga acccgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt
     2761 ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta
     2821 ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg
     2881 ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc
     2941 ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca
     3001 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     3061 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     3121 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     3181 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     3241 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     3301 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     3361 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     3421 cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     3481 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     3541 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     3601 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     3661 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag
     3721 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg
     3781 ggagcttgta tatccatttt cggatctgat cagcacgtgt tgacaattaa tcatcggcat
     3841 agtatatcgg catagtataa tacgacaagg tgaggaacta aaccatggcc aagttgacca
     3901 gtgccgttcc ggtgctcacc gcgcgcgacg tcgccggagc ggtcgagttc tggaccgacc
     3961 ggctcgggtt ctcccgggac ttcgtggagg acgacttcgc cggtgtggtc cgggacgacg
     4021 tgaccctgtt catcagcgcg gtccaggacc aggtggtgcc ggacaacacc ctggcctggg
     4081 tgtgggtgcg cggcctggac gagctgtacg ccgagtggtc ggaggtcgtg tccacgaact
     4141 tccgggacgc ctccgggccg gccatgaccg agatcggcga gcagccgtgg gggcgggagt
     4201 tcgccctgcg cgacccggcc ggcaactgcg tgcacttcgt ggccgaggag caggactgac
     4261 acgtgctacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg
     4321 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg
     4381 cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     4441 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     4501 atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt
     4561 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     4621 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaacttaca ttaattgcgt
     4681 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     4741 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     4801 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     4861 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     4921 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     4981 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     5041 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     5101 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     5161 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     5221 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     5281 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     5341 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     5401 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     5461 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     5521 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     5581 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     5641 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     5701 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     5761 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     5821 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     5881 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     5941 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     6001 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     6061 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     6121 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     6181 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     6241 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     6301 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     6361 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     6421 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     6481 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     6541 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     6601 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     6661 aaaataaaca aataggggtt ccgcgcacat ttccccga
//
