LOCUS       sequence-296450-        8898 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid contains mScarlet with CLASP. This consists of the plasma 
            membrane anchor (pm-LOVTRAP) and the cargo (Zdk-mScarlet-yeLANS). 
            CLASP modulates nuclear localization of mScarlet in response to bl.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8898)
  AUTHORS   Chen SY, Osimiri LC, Chevalier M, Bugaj LJ, Nguyen TH, Greenstein 
            RA, Ng AH, Stewart-Ornstein J, Neves LT, El-Samad H
  TITLE     Optogenetic Control Reveals Differential Promoter Interpretation of 
            Transcription Factor Nuclear Translocation Dynamics.
  JOURNAL   Cell Syst. 2020 Sep 4. pii: S2405-4712(20)30293-3. doi: 
            10.1016/j.cels.2020.08.009.
  PUBMED    32898473
REFERENCE   2  (bases 1 to 8898)
  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"; journalName: "Cell Syst. 
            2020 Sep 4. pii: S2405-4712(20)30293-3. doi: 
            10.1016/j.cels.2020.08.009."
COMMENT     Sequence Label: pCLASP_mScarlet_CLASP_RGS2membrane
FEATURES             Location/Qualifiers
     source          1..8898
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      2907..3094
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2962..2986)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     CDS             5858..6661
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     CDS             complement(7303..8118)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(7303..8118)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     8026..8045
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     8110..8129
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      join(8343..8898,1..33)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8832..8851
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
       61 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
      121 gtggataacc gtgcggccgc caatgctaaa ttcgagtgaa acacaggaag atcagaaaat
      181 cctcatttca tccatattaa caataatttc aaatgtttat ttgcattatt tgaaactagg
      241 caagacaagc aacgaaacgt ttttgaaaat tttgagtatt ttcaataaat ttgtagagga
      301 ctcagatatt gaaaaaaagc tacagcaatt aatacttgat aagaagagta ttgagaaggg
      361 caacggttca tcatctcatg gatctgcaca tgaacaaaca ccagagtcaa acgacgttga
      421 aattgaggct actgcgccaa ttgatgacaa tacagacgat gataacaaac cgaagttatc
      481 tgatgtagaa aaggattaaa gatgctaaga gatagtgatg atatttcata aataatgtaa
      541 ttctatatat gttaattacc ttttttgcga ggcatattta tggtgaagga taagttttga
      601 ccatcaaaga aggttaatgt ggctgtggtt tcagggtcca taaagcccac atggataaca
      661 ttacccctga attcgcatct agactgaact ggccgataat tgcagacgaa cgaagaggat
      721 gtccaatatt ttttttaagg aataaggata cttcaagact agattccccc ctgcattccc
      781 atcagaaccg taaaccttgg cgctttcctt gggaagtatt caagaagtgc cttgtccggt
      841 ttctgtggct cacaaaccag cgcgcccgat atggctttct tttcacttat gaatgtacca
      901 gtacgggaca attagaacgc tcctgtaaca atctctttgc aaatgtgggg ttacattcta
      961 accatgtcac actgctgacg aaattcaaag taaaaaaaaa tgggaccacg tcttgagaac
     1021 gatagatttt ctttatttta cattgaacag tcgttgtctc agcgcgcttt atgttttcat
     1081 tcatacttca tattataaaa taacaaaaga agaatttcat attcacgccc aagaaatcag
     1141 gctgctttcc aaatgcaatt gacacttcat tagccatcac acaaaactct ttcttgctgg
     1201 agcttctttt aaaaaagacc tcagtacacc aaacacgtta cccgacctcg ttattttacg
     1261 acaactatga taaaattctg aagaaaaaat aaaaaaattt tcatacttct tgcttttatt
     1321 taaaccattg aatgatttct tttgaacaaa actacctgtt tcaccaaagg aaatagaaag
     1381 aaaaaatcaa ttagaagaaa acaaaaaaca aaagatctat gggttctgtt gataataagt
     1441 ttaataaaga gaaaacaaga gctggagccg aaatacactc tttgcctaac ttaaatgtag
     1501 aacagaagtt cgcttttata gtatctttgt ttgatgatcc atcacaatct gccaatttgt
     1561 tggcagaggc taagaaatta aacgacgctc aagcaccaaa aggttctgta tcaaaaggcg
     1621 aggcggtcat taaagagttc atgaggttta aagtccatat ggaaggaagc atgaatggtc
     1681 acgagtttga gatagaggga gagggtgagg gaaggccgta cgaagggacg caaactgcga
     1741 agttgaaggt aacgaaaggc ggccctctgc cattctcttg ggacatcctt tcaccgcaat
     1801 ttatgtacgg ctctagggcc ttcacaaaac accccgccga tattccggat tactataaac
     1861 aaagttttcc tgagggcttc aagtgggaaa gagtaatgaa cttcgaggat gggggagctg
     1921 tgacggttac tcaagacact agcctggagg acggtactct tatatacaag gtaaaactaa
     1981 gagggacgaa ttttccgcca gacggccccg tgatgcaaaa aaaaactatg gggtgggaag
     2041 catccaccga gaggctgtac cccgaagacg gggtcctgaa gggtgatatt aaaatggcat
     2101 taagattaaa agatggcggg agatacttag ctgacttcaa gactacatat aaagccaaaa
     2161 aacctgtaca gatgcctgga gcttacaatg tggatcgtaa gctggatatt acatcccaca
     2221 acgaagatta taccgtggtg gagcagtacg agcgttcaga ggggcgtcac tccaccgggg
     2281 gtatggatga attatacaag agcggtctta gaagcagggc gcaagcgtca aattcagcgg
     2341 tagatggaac agcaggtcca ggtagtactg gcagcagagg atcccccgaa ttcggcattg
     2401 atctgagcgg cctgaccctg cagggttctg gaagtgggtc gttggctact acacttgaac
     2461 gtattgagaa gaactttgtc attactgacc caagattgcc agataatccc attatattcg
     2521 cgtccgatag tttcttgcag ttgacagaat atagccgtga agaaattttg ggaagaaact
     2581 gcaggtttct acaaggtcct gaaactgatc gcgcgacagt gagaaaaatt agagatgcca
     2641 tagataacca aacagaggtc actgttcagc tgattaatta tacaaagagt ggtaaaaagt
     2701 tctggaacct ctttcacttg cagcctatgc gagatcagaa gggagatgtc cagtacttta
     2761 ttggggttca gttggatgga actgagcatg tccgagatgc tgccgagaga gagggagtca
     2821 tgctgattaa gaaaactgca gaaaatattg atgctgctgc taagagatct tggtctatgg
     2881 ctttcgagct cggatcctaa ctcgaggcga atttcttatg atttatgatt tttattatta
     2941 aataagttat aaaaaaaata agtgtataca aattttaaag tgactcttag gttttaaaac
     3001 gaaaattctt attcttgagt aactctttcc tgtaggtcag gttgctttct caggtatagc
     3061 atgaggtcgc tcttattgac cacacctcta ccggcatgcc gagcaaatgc ctgcaaatcg
     3121 ctccccattt cgctgccaaa ccagatgtca acacagctac aacgaagagg atgtccaata
     3181 ttttttttaa ggaataagga tacttcaaga ctagattccc ccctgcattc ccatcagaac
     3241 cgtaaacctt ggcgctttcc ttgggaagta ttcaagaagt gccttgtccg gtttctgtgg
     3301 ctcacaaacc agcgcgcccg atatggcttt cttttcactt atgaatgtac cagtacggga
     3361 caattagaac gctcctgtaa caatctcttt gcaaatgtgg ggttacattc taaccatgtc
     3421 acactgctga cgaaattcaa agtaaaaaaa aatgggacca cgtcttgaga acgatagatt
     3481 ttctttattt tacattgaac agtcgttgtc tcagcgcgct ttatgttttc attcatactt
     3541 catattataa aataacaaaa gaagaatttc atattcacgc ccaagaaatc aggctgcttt
     3601 ccaaatgcaa ttgacacttc attagccatc acacaaaact ctttcttgct ggagcttctt
     3661 ttaaaaaaga cctcagtaca ccaaacacgt tacccgacct cgttatttta cgacaactat
     3721 gataaaattc tgaagaaaaa ataaaaaaat tttcatactt cttgctttta tttaaaccat
     3781 tgaatgattt cttttgaaca aaactacctg tttcaccaaa ggaaatagaa agaaaaaatc
     3841 aattagaaga aaacaaaaaa caaaagatct atgaagagaa cattgttgaa ggattggaaa
     3901 accagattat catacttctt acagaactca tctacacctg gtaagccaaa gacaggtaag
     3961 aaatctaaac aacaaggcgg tggcggaagt ggcggaggag gttcgggcgg aagtggttct
     4021 gctgaaggtt ctgttgctag acaaccagac ttgttgactt gtgacgacga accaatccac
     4081 atcccaggtg ctatccaacc acacggtttg ttgttggctt tggctgctga catgactatc
     4141 gttgctggtt ctgacaactt gccagaattg actggtttgg ctatcggtgc tttgatcggt
     4201 agatctgctg ctgacgtttt cgactctgaa actcacaaca gattgactat cgctttggct
     4261 gaaccaggtg ctgctgttgg tgctccaatc actgttggtt tcactatgag aaaggacgct
     4321 ggtttcatcg gttcttggca cagacacgac caattgatct tcttggaatt ggaaccacca
     4381 caaagagatg ttgctgaacc acaagctttc ttcagaagaa ctaactctgc tatcagaaga
     4441 ttgcaagctg ctgaaacttt ggaatctgct tgtgctgctg ctgctcaaga agttagaaag
     4501 atcactggtt tcgacagagt tatgatctac agattcgctt ctgacttctc tggtgaagtt
     4561 atcgctgaag acagatgtgc tgaagttgaa tctaagttgg gtttgcacta cccagcttct
     4621 actgttccag ctcaagctag aagattgtac actatcaacc cagttagaat catcccagac
     4681 atcaactaca gaccagttcc agttactcca gacttgaacc cagttactgg tagaccaatc
     4741 gacttgtctt tcgctatctt gagatctgtt tctccagttc acttggaatt catgagaaac
     4801 atcggtatgc acggtactat gtctatctct atcttgagag gtgaaagatt gtggggtttg
     4861 atcgtttgtc accacagaac tccatactac gttgacttgg acggtagaca agcttgtgaa
     4921 ttggttgctc aagttttggc ttggcaaatc ggtgttatgg aagaaggatc cttagctact
     4981 acgctagagc gtatagaaaa aaatttcgtg atcaccgatc caagattgcc ggacaaccca
     5041 atcatatttg cttccgattc ctttttacaa ctgaccgaat acagtagaga ggagattctg
     5101 ggtaggaatt gcaggttcct tcagggtccc gagactgatc gtgcaacagt taggaaaatt
     5161 agagatgcga tagacagcca gacagaagtt accgtgcagt tgatcaatta taccaagagc
     5221 ggaaaaaaat tctggaactt atttcattta cagccaatgc gtgaccagaa gggcgacgta
     5281 cagtatttca ttggtgtgca actagacggg acagagcacg tacgtgacgc agcagaaaga
     5341 gagggcgtga tgttgataaa aaaaactgca gagaatatcg acgaggctgc gaaagagcta
     5401 taatggcatt gaattgaatt gaaatcgata gatcaatttt tttcttttct ctttccccat
     5461 cctttacgct aaaataatag tttattttat tttttgaata ttttttattt atatacgtat
     5521 atatagacta ttatttatct tttaatgatt attaagattt ttattaaaaa aaaattcgct
     5581 cctcttttaa tgcctttatg cagttttttt ttcccattcg atatttctat gtgctgagca
     5641 gttacagaga tgttacgaac cactagtgca ctgcagtaca cggtttcctt gaaatttttt
     5701 tgattcggta atctccgaac agaaggaaga acgaaggaag gagcacagac ttagattggt
     5761 atatatacgc atatgtagtg ttgaagaaac atgaaattgc ccagtattct taacccaact
     5821 gcacagaaca aaaacgtgca ggaaacgaag ataaatcatg tcgaaagcta catataagga
     5881 acgtgctgct actcatccta gtcctgttgc tgccaagcta tttaatatca tgcacgaaaa
     5941 gcaaacaaac ttgtgtgctt cattggatgt tcgtaccacc aaggaattac tggagttagt
     6001 tgaagcatta ggtcccaaaa tttgtttact aaaaacacat gtggatatct tgactgattt
     6061 ttccatggag ggcacagtta agccgctaaa ggcattatcc gccaagtaca attttttact
     6121 cttcgaagac agaaaatttg ctgacattgg taatacagtc aaattgcagt actctgcggg
     6181 tgtatacaga atagcagaat gggcagacat tacgaatgca cacggtgtgg tgggcccagg
     6241 tattgttagc ggtttgaagc aggcggcaga agaagtaaca aaggaaccta gaggcctttt
     6301 gatgttagca gaattgtcat gcaagggctc cctatctact ggagaatata ctaagggtac
     6361 tgttgacatt gcgaagagcg acaaagattt tgttatcggc tttattgctc aaagagacat
     6421 gggtggaaga gatgaaggtt acgattggtt gattatgaca cccggtgtgg gtttagatga
     6481 caagggagat gcattgggtc aacagtatag aaccgtggat gatgtggttt ctacaggatc
     6541 tgacattatt attgttggaa gaggactatt tgcaaaggga agggatgcta aggtagaggg
     6601 tgaacgttac agaaaagcag gctgggaagc atatttgaga agatgcggcc agcaaaacta
     6661 aaaaactgta ttataagtaa atgcatgtat actaaactca caaattagag cttcaattta
     6721 attatatcag ttattacccg agtagagcac ttgaatccac tgccccggga atctcggtcg
     6781 taatgatttc tataatgacg aaaaaaaaaa aattggaaag aaaaagcttc atggccttta
     6841 taaaaaggaa ctatccaata cctcgccaga accaagtaac agtattttac ggggcacaaa
     6901 tcaagaacaa taagacagga ctgtaaagat ggacgcattg aactccaaag aacaacaaga
     6961 gttccaaaaa gtagtggaac aaaagcaaat gaaggatttc atgcgtttgt actctaatct
     7021 ggtagaaaga tgtttcacag actgtgtcaa tgacttcaca acatcaaagc taaccaataa
     7081 ggaacaaaca tgcatcatga agtgctcaga aaagttcttg aagcatagcg aacgtgtagg
     7141 gcagcgtttc caagaacaaa acgctgcctt gggacaaggc ttgggccgat aaggtgtact
     7201 ggcgtatata tatctaatta tgtatctctg gtgtagccca tttttagcat gtaaatataa
     7261 agaccgagcg gccgcgtgtt acaaccaatt aaccaattct gattagaaaa actcatcgag
     7321 catcaaatga aactgcaatt tattcatatc aggattatca ataccatatt tttgaaaaag
     7381 ccgtttctgt aatgaaggag aaaactcacc gaggcagttc cataggatgg caagatcctg
     7441 gtatcggtct gcgattccga ctcgtccaac atcaatacaa cctattaatt tcccctcgtc
     7501 aaaaataagg ttatcaagtg agaaatcacc atgagtgacg actgaatccg gtgagaatgg
     7561 caaaagctta tgcatttctt tccagacttg ttcaacaggc cagccattac gctcgtcatc
     7621 aaaatcactc gcatcaacca aaccgttatt cattcgtgat tgcgcctgag cgaggcgaaa
     7681 tacgcgatcg ctgttaaaag gacaattaca aacaggaatc gaatgcaacc ggcgcaggaa
     7741 cactgccagc gcatcaacaa tattttcacc tgaatcagga tattcttcta atacctggaa
     7801 tgctgttttc ccggggatcg cagtggtgag taaccatgca tcatcaggag tacggataaa
     7861 atgcttgatg gtcggaagag gcataaattc cgtcagccag tttagtctga ccatctcatc
     7921 tgtaacatca ttggcaacgc tacctttgcc atgtttcaga aacaactctg gcgcatcggg
     7981 cttcccatac aatcgataga ttgtcgcacc tgattgcccg acattatcgc gagcccattt
     8041 atacccatat aaatcagcat ccatgttgga atttaatcgc ggcctggagc aagacgtttc
     8101 ccgttgaata tggctcataa caccccttgt attactgttt atgtaagcag acagttttat
     8161 tgttcatgat gatatatttt tatcttgtgc aatgtaacat cagagatttt gagacacaac
     8221 gtggctttgt tgaataaatc gaacttttgc tgagttgaag gatcagtcat gaccaaaatc
     8281 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     8341 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     8401 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     8461 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     8521 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     8581 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     8641 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     8701 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     8761 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     8821 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     8881 cttgagcgtc gatttttg
//
