LOCUS       sequence-363934-        6077 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For the mammalian expression of the sleeping chironomid protein 
            pvLEA22mer_mutant_3 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 6077)
  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 6077)
  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_mutant_3_mCh-SspB (pBS1080)
FEATURES             Location/Qualifiers
     source          1..6077
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        23..127
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(128..196,197..988)
                     /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     complement(346..365)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1159..1747
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1648..1667
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1901..1918
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        join(2071..2076,2077..2094,2095..2101)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2109..2125
                     /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)."
     primer_bind     2114..2136
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2133..2149
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2133..2149
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    2198..2319
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2209..2228)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2263..2282
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(2449..2823)
                     /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"
     promoter        complement(2842..2889)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(2937..3266)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2951..3086
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3005..3024)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     3251..3271
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(3280..3708)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3390..3411)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3602..3621
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    3754..3978
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3967..3984
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(4004..4333)
                     /codon_start=1
                     /product="SspB from Haemophilus influenzae, with the 
                     mutations Y11K and A15E to disrupt the natural dimerization
                     of the protein"
                     /label=SspB nano
                     /note="dimerizes with iLID at high affinity to create iLID 
                     nano (Guntas et al., 2015)"
                     /translation="SSPKRPKLLREYYDWLVDNSFTPYLVVDATYLGVNVPVEYVKDGQ
                     IVLNLSASATGNLQLTNDFIQFNARFKGVSRELYIPMGAALAIYARENGDGVMFEPEEI
                     YDELNI"
     regulatory      5161..5170
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(5187..5206)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5188..5206)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(5251..5454)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(5280..5300)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        5455..5834
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     6006..6025
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
       61 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
      121 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
      181 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
      241 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
      301 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
      361 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
      421 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
      481 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
      541 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
      601 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
      661 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
      721 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
      781 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
      841 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
      901 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
      961 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     1021 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     1081 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     1141 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     1201 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     1261 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     1321 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     1381 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     1441 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     1501 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     1561 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     1621 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     1681 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     1741 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     1801 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
     1861 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
     1921 agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg
     1981 cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt
     2041 aagttagctc actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt
     2101 gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc
     2161 caagctctag ctagaggtcg acggtataca gacatgataa gatacattga tgagtttgga
     2221 caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt
     2281 gctttatttg taaccattat aagctgcaat aaacaagttg gggtgggcga agaactccag
     2341 catgagatcc ccgcgctgga ggatcatcca gccggcgtcc cggaaaacga ttccgaagcc
     2401 caacctttca tagaaggcgg cggtggaatc gaaatctcgt agcacgtgtc agtcctgctc
     2461 ctcggccacg aagtgcacgc agttgccggc cgggtcgcgc agggcgaact cccgccccca
     2521 cggctgctcg ccgatctcgg tcatggccgg cccggaggcg tcccggaagt tcgtggacac
     2581 gacctccgac cactcggcgt acagctcgtc caggccgcgc acccacaccc aggccagggt
     2641 gttgtccggc accacctggt cctggaccgc gctgatgaac agggtcacgt cgtcccggac
     2701 cacaccggcg aagtcgtcct ccacgaagtc ccgggagaac ccgagccggt cggtccagaa
     2761 ctcgaccgct ccggcgacgt cgcgcgcggt gagcaccgga acggcactgg tcaacttggc
     2821 catggtttag ttcctcacct tgtcgtatta tactatgccg atatactatg ccgatgatta
     2881 attgtcaaca cgtgctgatc agatccgaaa atggatatac aagctcccgg gagctttttg
     2941 caaaagccta ggcctccaaa aaagcctcct cactacttct ggaatagctc agaggcagag
     3001 gcggcctcgg cctctgcata aataaaaaaa attagtcagc catggggcgg agaatgggcg
     3061 gaactgggcg gagttagggg cgggatgggc ggagttaggg gcgggactat ggttgctgac
     3121 taattgagat gcatgctttg catacttctg cctgctgggg agcctgggga ctttccacac
     3181 ctggttgctg actaattgag atgcatgctt tgcatacttc tgcctgctgg ggagcctggg
     3241 gactttccac accctaactg acacacattc cacagaatta attcgcgtta aatttttgtt
     3301 aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag
     3361 aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga
     3421 acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg
     3481 aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc
     3541 ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg
     3601 aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc
     3661 gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtgg ggataccccc
     3721 tagagcccca gctggttctt tccgcctcag aagccataga gcccaccgca tccccagcat
     3781 gcctgctatt gtcttcccaa tcctccccct tgctgtcctg ccccacccca ccccccagaa
     3841 tagaatgaca cctactcaga caatgcgatg caatttcctc attttattag gaaaggacag
     3901 tgggagtggc accttccagg gtcaaggaag gcacggggga ggggcaaaca acagatggct
     3961 ggcaactaga aggcacagtc gaggctgatc agcgggttta accaatattc agctcgtcat
     4021 agatttcttc tggttcgaac atcacaccat cgccgttctc gcgagcgtaa atggccagag
     4081 cggcacccat cgggatatac agttcacgag acacgccctt aaagcgggcg ttgaactgga
     4141 taaaatcatt tgtcagttgc aggttgccgg tcgcacttgc agacagattc agcacgatct
     4201 gaccgtcttt cacatactcc acgggcacgt tcacgcccag gtatgtggca tccaccacca
     4261 gatatggggt aaagctgtta tcaaccagcc aatcgtaata ttcacgcagc agcttagggc
     4321 gtttcgggga gctacctcca gaggtaccac cggatccggc cgccttgtac aattcatcca
     4381 tgccccctgt gctatgtctg ccctcggccc gctcgtactg ttccacgatg gtatagtctt
     4441 cgttatggga tgtaatatcc agtttgatat tcacgttgta tgccccagga agctgaacgg
     4501 gtttcttggc cttataggtg gttttgacct cagcatcata atgtccccca tccttcagct
     4561 tgagtcgctg cttgatttct cccttcagtg ctccgtcctc gggatacatg cgttctgagc
     4621 tggcttccca gcccatagtt ttcttctgca tcacggggcc gtctgatggg aagttagtgc
     4681 cccgaagttt aaccttatag ataaattctc cgtcctgaag agagctgtcc tgggtaactg
     4741 tgactacacc gccatcctca aaattcatta cccgttccca cttgaaccct tcagggaagc
     4801 tcagtttcaa gtagtccgga atatcggcag ggtgcttaac gtaagcctta gagccgtaca
     4861 taaactgggg tgacaggatg tcccaggcga aaggcagggg gccacccttg gtcaccttca
     4921 gtttggccgt ctgtgtccct tcatagggtc gcccctcgcc ttcgccctcg atttcgaact
     4981 cgtgaccgtt cacagatccc tccatatgca ccttgaacct cataaattcc ttgattatag
     5041 ccatgttatc ctcctcgcct ttagacacca ttccgccgga tccgccgcct ccggactccc
     5101 cggctttctc cacggtggcg tcctttgcct ctcctgcctt ctccacggtt gcgtccttgg
     5161 ccatggtggc tagccagctt gggtctccct atagtgagtc gtattaattt cgataagcca
     5221 gtaagcagtg ggttctctag ttagccagag agctctgctt atatagacct cccaccgtac
     5281 acgcctaccg cccatttgcg tcaatggggc ggagttgtta cgacattttg gaaagtcccg
     5341 ttgattttgg tgccaaaaca aactcccatt gacgtcaatg gggtggagac ttggaaatcc
     5401 ccgtgagtca aaccgctatc cacgcccatt gatgtactgc caaaaccgca tcaccatggt
     5461 aatagcgatg actaatacgt agatgtactg ccaagtagga aagtcccata aggtcatgta
     5521 ctgggcataa tgccaggcgg gccatttacc gtcattgacg tcaatagggg gcgtacttgg
     5581 catatgatac acttgatgta ctgccaagtg ggcagtttac cgtaaatact ccacccattg
     5641 acgtcaatgg aaagtcccta ttggcgttac tatgggaaca tacgtcatta ttgacgtcaa
     5701 tgggcggggg tcgttgggcg gtcagccagg cgggccattt accgtaagtt atgtaacgcg
     5761 gaactccata tatgggctat gaactaatga ccccgtaatt gattactatt aataactagt
     5821 caataatcaa tgtcaacgcg tatatctggc ccgtacatcg cgaagcagcg caaaacgcct
     5881 aaccctaagc agattcttca tgcaattgtc ggtcaagcct tgccttgttg tagcttaaat
     5941 tttgctcgcg cactactcag cgacctccaa cacacaagca gggagcagat actggcttaa
     6001 ctatgcggca tcagagcaga ttgtactgag agtgcaccat aggggatcgg gagatctccc
     6061 gatccgtcga cgtcagg
//
