LOCUS       sequence-363927-        6110 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For the mammalian expression of the tardigrade protein 
            CAHS2_PARRC_98-130 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 6110)
  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 6110)
  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: CAHS2_PARRC_98-130_mCh-SspB (pBS1077)
FEATURES             Location/Qualifiers
     source          1..6110
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(51..70)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        242..621
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        622..825
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     776..796
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     870..889
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        870..888
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      906..915
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1776..2105
                     /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"
     primer_bind     complement(2125..2142)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2131..2355
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2401..2829
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2488..2507)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2698..2719
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2838..2858)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2843..3172
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3023..3158
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3085..3104
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3220..3267
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3286..3660
                     /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    3790..3911
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3827..3846)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3881..3900
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3960..3976)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3960..3976)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3973..3995)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3984..4000
                     /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(4008..4014,4015..4032,4033..4038))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(4191..4208)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4362..4950)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4442..4461)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5121..5912,5913..5981))
                     /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     5744..5763
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5982..6086)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tgacgtcgac ggatcgggag atctcccgat cccctatggt gcactctcag tacaatctgc
       61 tctgatgccg catagttaag ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag
      121 tagtgcgcga gcaaaattta agctacaaca aggcaaggct tgaccgacaa ttgcatgaag
      181 aatctgctta gggttaggcg ttttgcgctg cttcgcgatg tacgggccag atatacgcgt
      241 tgacattgat tattgactag ttattaatag taatcaatta cggggtcatt agttcatagc
      301 ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc
      361 aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg
      421 actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat
      481 caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc
      541 tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta catctacgta
      601 ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag
      661 cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt
      721 tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa
      781 atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctctctg gctaactaga
      841 gaacccactg cttactggct tatcgaaatt aatacgactc actataggga gacccaagct
      901 ggctagccac catggccgcc aagactagag agaacgagat gctgggccag cagtacgaga
      961 aagagttgga gcggaagtcc gaggcttaca gaaaacacca ggaggtggag gcctccggag
     1021 gcggcggatc cggcggaatg gtgtctaaag gcgaggagga taacatggct ataatcaagg
     1081 aatttatgag gttcaaggtg catatggagg gatctgtgaa cggtcacgag ttcgaaatcg
     1141 agggcgaagg cgaggggcga ccctatgaag ggacacagac ggccaaactg aaggtgacca
     1201 agggtggccc cctgcctttc gcctgggaca tcctgtcacc ccagtttatg tacggctcta
     1261 aggcttacgt taagcaccct gccgatattc cggactactt gaaactgagc ttccctgaag
     1321 ggttcaagtg ggaacgggta atgaattttg aggatggcgg tgtagtcaca gttacccagg
     1381 acagctctct tcaggacgga gaatttatct ataaggttaa acttcggggc actaacttcc
     1441 catcagacgg ccccgtgatg cagaagaaaa ctatgggctg ggaagccagc tcagaacgca
     1501 tgtatcccga ggacggagca ctgaagggag aaatcaagca gcgactcaag ctgaaggatg
     1561 ggggacatta tgatgctgag gtcaaaacca cctataaggc caagaaaccc gttcagcttc
     1621 ctggggcata caacgtgaat atcaaactgg atattacatc ccataacgaa gactatacca
     1681 tcgtggaaca gtacgagcgg gccgagggca gacatagcac agggggcatg gatgaattgt
     1741 acaaggcggc cggatccggt ggtacctctg gaggtagctc cccgaaacgc cctaagctgc
     1801 tgcgtgaata ttacgattgg ctggttgata acagctttac cccatatctg gtggtggatg
     1861 ccacatacct gggcgtgaac gtgcccgtgg agtatgtgaa agacggtcag atcgtgctga
     1921 atctgtctgc aagtgcgacc ggcaacctgc aactgacaaa tgattttatc cagttcaacg
     1981 cccgctttaa gggcgtgtct cgtgaactgt atatcccgat gggtgccgct ctggccattt
     2041 acgctcgcga gaacggcgat ggtgtgatgt tcgaaccaga agaaatctat gacgagctga
     2101 atattggtta aacccgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg
     2161 tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct
     2221 aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg
     2281 gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg
     2341 cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc
     2401 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     2461 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     2521 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     2581 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
     2641 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     2701 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
     2761 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
     2821 acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     2881 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
     2941 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
     3001 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     3061 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga
     3121 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc
     3181 gggagcttgt atatccattt tcggatctga tcagcacgtg ttgacaatta atcatcggca
     3241 tagtatatcg gcatagtata atacgacaag gtgaggaact aaaccatggc caagttgacc
     3301 agtgccgttc cggtgctcac cgcgcgcgac gtcgccggag cggtcgagtt ctggaccgac
     3361 cggctcgggt tctcccggga cttcgtggag gacgacttcg ccggtgtggt ccgggacgac
     3421 gtgaccctgt tcatcagcgc ggtccaggac caggtggtgc cggacaacac cctggcctgg
     3481 gtgtgggtgc gcggcctgga cgagctgtac gccgagtggt cggaggtcgt gtccacgaac
     3541 ttccgggacg cctccgggcc ggccatgacc gagatcggcg agcagccgtg ggggcgggag
     3601 ttcgccctgc gcgacccggc cggcaactgc gtgcacttcg tggccgagga gcaggactga
     3661 cacgtgctac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
     3721 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
     3781 gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     3841 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     3901 aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg
     3961 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     4021 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaacttac attaattgcg
     4081 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     4141 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     4201 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     4261 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     4321 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     4381 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     4441 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     4501 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     4561 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     4621 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     4681 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     4741 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     4801 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     4861 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     4921 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     4981 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     5041 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     5101 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     5161 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     5221 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     5281 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     5341 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     5401 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     5461 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     5521 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     5581 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     5641 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     5701 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     5761 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     5821 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     5881 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     5941 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     6001 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     6061 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
//
