LOCUS       sequence-254569-        6829 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Redshifted FRET-readout of Stathmin/OP18 interaction with 
            alpha-tubulin.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6829)
  AUTHORS   Van Geel O, Cheung S, Gadella TWJ
  TITLE     Combining optogenetics with sensitive FRET imaging to monitor local 
            microtubule manipulations.
  JOURNAL   Sci Rep. 2020 Apr 7;10(1):6034. doi: 10.1038/s41598-020-62874-3.
  PUBMED    32265472
REFERENCE   2  (bases 1 to 6829)
  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: "Sci Rep."; 
            date: "2020-04-7"; volume: "10(1):6034. doi"; pages: " 
            10.1038/s41598-020-62874-3"
COMMENT     Sequence Label: sYFP2-OP18-mScI
FEATURES             Location/Qualifiers
     source          1..6829
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        306..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     711..733
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     795..814
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     promoter        856..974
                     /label=rhaB promoter
                     /note="promoter of the E. coli rhaBAD operon, conferring 
                     tight induction with L-rhamnose and repression with 
                     D-glucose in the presence of RhaR and RhaS (Giacalone et 
                     al., 2006)"
     CDS             1049..1066
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1076..1093
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     regulatory      1112..1121
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1118..1120,1121..1123,1124..1834)
                     /codon_start=1
                     /product="monomeric optimized variant of EYFP, identical to
                     mVenus but with L68V (Kremers et al., 2006)"
                     /label=SYFP2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1163..1184)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1424..1443)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1771..1792
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2312..3004
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet-I
                     /note="matures faster than mScarlet"
                     /translation="VSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFSWDILSPQFMYGSRAFIKHPADIPDYYKQSFPEGFKWERVMNFEDGGA
                     VTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIKM
                     ALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRHS
                     TGGMDELYK"
     primer_bind     complement(2442..2460)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2710..2729
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(3094..3113)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3159..3214
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3213..3232)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     terminator      3302..3349
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_recomb     3362..4067
                     /label=baculovirus recombination region (ORF1629)
                     /note="contains part of ORF1629"
     3'UTR           3450..3822
                     /label=polyhedrin 3' UTR
                     /note="3' UTR from the Autographa californica 
                     nucleopolyhedrovirus polyhedrin gene (Suzuki et al., 2006)"
     CDS             complement(3479..4067)
                     /codon_start=2
                     /product="baculovirus capsid-associated protein"
                     /label=ORF1629
                     /note="required for viral replication"
                     /translation="IPKSSTTNLIADVLADTINRRRVAMAKSSSEATSNDEGWDDDDNR
                     PNKANTPDVKYVQALFNVFTSSQLYTNDSDERNTKAHNILNDVEPLLQNKTQTNIDKAR
                     LLLQDLASFVALSENPLDSPAIGSEKQPLFETNRNLFYKSIEDLIFKFRYKDAENHLIF
                     ALTYHPKDYKFNELLKYVQQLSVNQQRTESSA"
     protein_bind    complement(4076..4109)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     rep_origin      complement(4277..4864)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4356..4375)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4877..5692)
                     /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
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNGDRVFRLAQAQSRMNNGLVGA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     5600..5619
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     misc_recomb     5935..6762
                     /label=baculovirus recombination region (lef2/ORF603)
                     /note="contains ORF603 and part of lef2"
     CDS             5935..6059
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(6097..6702)
                     /codon_start=1
                     /product="baculovirus ORF603 protein"
                     /label=ORF603
                     /translation="MAVIFNNKQLLADNSIEKGGELFLFNGSYNILESYVNPVLLKNGV
                     IELEEAAYYAGNILYKTDDPKFIDYINLIIKATHSEELPENSTVVNYRKTMRSGTIHPI
                     KKDIYIYDNKKFTLYDRYIYGYDNNYVNFYEEKNEKEKEYEEEDDKASSLCENKIILSQ
                     INCESFENDFKYYLSDYNYAFSIIDNTTNVLVAFGLYR"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtccgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta cgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catgctgatg cggttttggc agtacaccaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta ataaccccgc cccgttgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagacgtcg tttagtgaac cgtcagatca ctagatgctt
      541 tattgcggta gtttatcaca gttaaattgc taacgccagt ctcgaactta acgtgcagaa
      601 gttggtcgtg aggcactggg caggtaagta tcgggccctt tgtgcggggg gagcggctcg
      661 gggctgtccg cggggggacg gctgccttcg ggggggacgg ggcagggcgg ggttcggctt
      721 ctggcgtgtg accggcggct ctagagcctc tgctaaccat gttcatgcct tcttcttttt
      781 cctacagctc ctgggcaacg tgctggttat tgtgctgtct catcattttg gcaaagaatt
      841 ggatcggacc gaaatcacca caattcagca aattgtgaac atcatcacgt tcatctttcc
      901 ctggttgcca aaggcccatt ttcctgtcag taacgagaag gtcgcgaatt caggcgcttt
      961 ttagactggt cgtagggaga ccacaacggt ttccctctag aaataatttt gtttaactat
     1021 aagaaggaga tataccatgc ggggttctca tcatcaccac catcacagca gcggcctggt
     1081 gccgcgcggc agctcgctag cgctaccggt cgccaccatg gtgagcaagg gcgaggagct
     1141 gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt
     1201 cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagctgat
     1261 ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgggctacgg
     1321 cgtgcagtgc ttcgcccgct accccgacca catgaagcag cacgacttct tcaagtccgc
     1381 catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa
     1441 gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg
     1501 catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag
     1561 ccacaacgtc tatatcaccg ccgacaagca gaagaacggc atcaaggcca acttcaagat
     1621 ccgccacaac atcgaggacg gcggcgtgca gctcgccgac cactaccagc agaacacccc
     1681 catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagctacc agtccaagct
     1741 gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc
     1801 cgggatcact ctcggcatgg acgagctgta caagtccgga ctcagatcta cgatggcatc
     1861 ttctgatatt caggtgaaag agctggagaa gcgcgcttca ggccaggctt ttgagctgat
     1921 tctcagccct cggtcaaaag aatctgtccc cgatttcccc ctttcccccc caaagaagaa
     1981 ggacctttcc ctggaggaaa tccagaagaa attagaagct gcagaagaaa gacgcaagtc
     2041 tcatgaggcg gaagtcttga agcagctcgc ggagaagcgg gagcatgaga aggaggtgct
     2101 ccagaaagcc atcgaggaga acaacaactt cagcaagatg gcggaggaga agctgaccca
     2161 caaaatggag gctaacaaag agaaccggga ggcgcagatg gcggccaagc tggagcgctt
     2221 gcgagagaag gacaagcacg tggaagaggt gcggaagaac aaagaatcca aagaccccgc
     2281 ggatgagact gaggctgacg gatccggttc agtgagcaag ggcgaggcag tgatcaagga
     2341 gttcatgcgg ttcaaggtgc acatggaggg ctccatgaac ggccacgagt tcgagatcga
     2401 gggcgagggc gagggccgcc cctacgaggg cacccagacc gccaagctga aggtgaccaa
     2461 gggtggcccc ctgcccttct cctgggacat cctgtcccct cagttcatgt acggctccag
     2521 ggccttcatc aagcaccccg ccgacatccc cgactactat aagcagtcct tccccgaggg
     2581 cttcaagtgg gagcgcgtga tgaacttcga ggacggcggc gccgtgaccg tgacccagga
     2641 cacctccctg gaggacggca ccctgatcta caaggtgaag ctccgcggca ccaacttccc
     2701 tcctgacggc cccgtaatgc agaagaagac aatgggctgg gaagcgtcca ccgagcggtt
     2761 gtaccccgag gacggcgtgc tgaagggcga cattaagatg gccctgcgcc tgaaggacgg
     2821 cggccgctac ctggcggact tcaagaccac ctacaaggcc aagaagcccg tgcagatgcc
     2881 cggcgcctac aacgtcgacc gcaagttgga catcacctcc cacaacgagg actacaccgt
     2941 ggtggaacag tacgaacgct ccgagggccg ccactccacc ggcggcatgg acgagctgta
     3001 caaatgagcg gccgccctca ggtgcaggct gcctatcaga aggtggtggc tggtgtggcc
     3061 aatgccctgg ctcacaaata ccactgagat cgatcttttt ccctctgcca aaaattatgg
     3121 ggacatcatg aagccccttg agcatctgac ttctggctaa taaaggaaat ttattttcat
     3181 tgcaatagtg tgttggaatt ttttgtgtct ctcactcgga aggacatatg ggagggcaaa
     3241 tcatttaaaa catcagaatg agtatttggt ttagagtttg gcaacatatg cccatatgta
     3301 actagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgc tgaaagcatg
     3361 cggaggaaat tctccttgaa gtttccctgg tgttcaaagt aaaggagttt gcaccagacg
     3421 cacctctgtt cactggtccg gcgtattaaa acacgataca ttgttattag tacatttatt
     3481 aagcgctaga ttctgtgcgt tgttgattta cagacaattg ttgtacgtat tttaataatt
     3541 cattaaattt ataatcttta gggtggtatg ttagagcgaa aatcaaatga ttttcagcgt
     3601 ctttatatct gaatttaaat attaaatcct caatagattt gtaaaatagg tttcgattag
     3661 tttcaaacaa gggttgtttt tccgaaccga tggctggact atctaatgga ttttcgctca
     3721 acgccacaaa acttgccaaa tcttgtagca gcaatctagc tttgtcgata ttcgtttgtg
     3781 ttttgttttg taataaaggt tcgacgtcgt tcaaaatatt atgcgctttt gtatttcttt
     3841 catcactgtc gttagtatac aattgactcg acgtaaacac gttaaataga gcttggacat
     3901 atttaacatc gggcgtgtta gctttattag gccgattatc gtcgtcgtcc caaccctcgt
     3961 cgttagaagt tgcttccgaa gacgattttg ccatagccac acgacgccta ttaattgtgt
     4021 cggctaacac gtccgcgatc aaatttgtag ttgagctttt tggaattgcg atcgcataac
     4081 ttcgtatagc atacattata cgaagttata agctcggaac gctgcgctcg gtcgttcggc
     4141 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     4201 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     4261 ccgcgttgct ggcgtttttc cataggctcc gccccctgac gagcatcaca aaaatcgacg
     4321 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     4381 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     4441 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     4501 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     4561 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     4621 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     4681 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     4741 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     4801 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     4861 tcaagaagat cctttgttag aaaaactcat cgagcatcaa gtgaaactgc aatttattca
     4921 tatcaggatt atcaatacca tatttttgaa aaagccgttt ctgtaatgaa ggagaaaact
     4981 caccgaggca gttccatagg atggcaagat cctggtatcg gtctgcgatt ccgactcgtc
     5041 caacatcaat acaacctatt aatttcccct cgtcaaaaat aaggttatca agtgagaaat
     5101 caccatgagt gacgactgaa tccggtgaga atggcaaaag cttatgcatt tctttccaga
     5161 cttgttcaac aggccagcca ttacgctcgt catcaaaatc actcgcacca accaaaccgt
     5221 tattcattcg tgattgcgcc tgagcgagac gaaatacgcg atcgccgtta aaaggacaat
     5281 tacaaacagg aatcgaatgc aaccggcgca ggaacactgc cagcgcatca acaatatttt
     5341 cacctgaatc aggatattct tctaatacct ggaatgctgt tttccctggg atcgcagtgg
     5401 tgagtaacca tgcatcatca ggagtacgga taaaatgctt gatggtcgga agaggcataa
     5461 attccgtcag ccagtttagc ctgaccatct catctgtaac atcattggca acgctacctt
     5521 tgccatgttt cagaaacaac tctggcgcat cgggcttccc atacaatcga tagattgtcg
     5581 cacctgattg cccgacatta tcgcgagccc atttataccc atataaatca gcatccatgt
     5641 tggaatttaa tcgcggcctc gagcaagacg tttcccgttg aatatggctc atactcttcc
     5701 tttttcaata ttattgaagc atttatcagg gttattgtct catgtccgcg cgtttcctgc
     5761 atcttttaat caaatcccaa gatgtgtata aacgcgccgg tatgtatagg aagaggttta
     5821 tactaaactg ttacattgca aacgtggttt cgtgtgccaa gtgtgaaaac cgatgtttaa
     5881 tcaaggctct gacgcatttc tacaaccacg actccaagtg tgtgggtgaa gtcatgcatc
     5941 ttttaatcaa atcccaagat gtgtataaac caccaaactg ccaaaaaatg aaaactgtcg
     6001 acaagctctg tccgtttgct ggcaactgca agggtctcaa tcctatttgt aattattgaa
     6061 taataaaaca attataaatg tcaaatttgt tttttattaa cgatacaaac caaacgcaac
     6121 aagaacattt gtagtattat ctataattga aaacgcgtag ttataatcgc tgaggtaata
     6181 tttaaaatca ttttcaaatg attcacagtt aatttgcgac aatataattt tattttcaca
     6241 taaactagac gccttgtcgt cttcttcttc gtattccttc tctttttcat ttttctcttc
     6301 ataaaaatta acatagttat tatcgtatcc atatatgtat ctatcgtata gagtaaattt
     6361 tttgttgtca taaatatata tgtctttttt aatggggtgt atagtaccgc tgcgcatagt
     6421 ttttctgtaa tttacaacag tgctattttc tggtagttct tcggagtgtg ttgctttaat
     6481 tattaaattt atataatcaa tgaatttggg atcgtcggtt ttatacaata tgttgccggc
     6541 atagtacgca gcttcttcta gttcaattac accatttttt agcagcaccg gattaacata
     6601 actttccaaa atgttgtacg aaccgttaaa caaaaacagt tcacctccct tttctatact
     6661 attgtctgcg agcagttgtt tgttgttaaa aataacagcc attgtaatga gacgcacaaa
     6721 ctaatatcac aaactggaaa tgtctatcaa tatatagttg ctgatggccg gcctattaat
     6781 agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccg
//
