LOCUS       sequence-254579-        6823 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Local FRET-readout of membrane-anchored Stathmin/OP18 interaction 
            with alpha-tubulin.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6823)
  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 6823)
  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: ROPY-2C2
FEATURES             Location/Qualifiers
     source          1..6823
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..204
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     155..175
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      243..252
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(249..251,252..254,255..965)
                     /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(294..315)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(555..574)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     902..923
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1573..1591)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1841..1860
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    3247..3368
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3284..3303)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3338..3357
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3375..3830)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3512..3533)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3724..3743
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3857..3961
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3963..4320
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3963..4154
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3986..4006)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4171..4306
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4355..5149
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             4355..5149
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4409..4428)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5019..5038
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5337..5356)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5381..5428
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5757..6345
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6246..6265
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        6520..6823
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
       61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      121 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      181 tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta gcgctaccgg
      241 tcgccaccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg
      301 agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg
      361 ccacctacgg caagctgacc ctgaagctga tctgcaccac cggcaagctg cccgtgccct
      421 ggcccaccct cgtgaccacc ctgggctacg gcgtgcagtg cttcgcccgc taccccgacc
      481 acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca
      541 ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg
      601 acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc
      661 tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcacc gccgacaagc
      721 agaagaacgg catcaaggcc aacttcaaga tccgccacaa catcgaggac ggcggcgtgc
      781 agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg
      841 acaaccacta cctgagctac cagtccaagc tgagcaaaga ccccaacgag aagcgcgatc
      901 acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt
      961 acaagtccgg actcagatct acgatggcat cttctgatat tcaggtgaaa gagctggaga
     1021 agcgcgcttc aggccaggct tttgagctga ttctcagccc tcggtcaaaa gaatctgtcc
     1081 ccgatttccc cctttccccc ccaaagaaga aggacctttc cctggaggaa atccagaaga
     1141 aattagaagc tgcagaagaa agacgcaagt ctcatgaggc ggaagtcttg aagcagctcg
     1201 cggagaagcg ggagcatgag aaggaggtgc tccagaaagc catcgaggag aacaacaact
     1261 tcagcaagat ggcggaggag aagctgaccc acaaaatgga ggctaacaaa gagaaccggg
     1321 aggcgcagat ggcggccaag ctggagcgct tgcgagagaa ggacaagcac gtggaagagg
     1381 tgcggaagaa caaagaatcc aaagaccccg cggatgagac tgaggctgac ggatccggtt
     1441 cagtgagcaa gggcgaggca gtgatcaagg agttcatgcg gttcaaggtg cacatggagg
     1501 gctccatgaa cggccacgag ttcgagatcg agggcgaggg cgagggccgc ccctacgagg
     1561 gcacccagac cgccaagctg aaggtgacca agggtggccc cctgcccttc tcctgggaca
     1621 tcctgtcccc tcagttcatg tacggctcca gggccttcat caagcacccc gccgacatcc
     1681 ccgactacta taagcagtcc ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg
     1741 aggacggcgg cgccgtgacc gtgacccagg acacctccct ggaggacggc accctgatct
     1801 acaaggtgaa gctcatcggc accaacttcc ctcctgacgg ccccgtaatg cagaagaaga
     1861 caatgggctg ggaagcgtcc accgagcggt tgtaccccga ggacggcgtg ctgaagggcg
     1921 acattaagat ggccctgcgc ctgaaggacg gcggccgcta cctggcggac ttcaagacca
     1981 cctacaaggc caagaagccc gtgcagatgc ccggcgccta caacgtcgac cgcaagttgg
     2041 acatcacctc ccacaacgag gactacaccg tggtggaaca gtacgaacgc tccgagggcc
     2101 gccactccac cggaagtagt tgcactgaac ccctaggcct gaaggataat accatcccca
     2161 acaagcagat cacagcctcc agctactaca aaacctgggg cctgagtgcc tttagctggt
     2221 ttccctacta cgcacgactg gataatcagg gcaagttcaa cgcctggacc gcccagacca
     2281 acagtgcctc tgagtggctg cagattgacc tgggctccca gaagcgagtc acgggcatca
     2341 tcacccaggg tgcccgagac tttggccaca ttcaatatgt ggctgcctac agggtggcct
     2401 atggtgatga tggtgtgacc tggactgagt acaaggaccc gggggcctca gaaagcaaga
     2461 ttttccctgg taacatggac aataattccc acaagaagaa catatttgag acgccgttcc
     2521 aggctcgctt cgtgcggatc cagcccgtgg cctggcacaa ccgtatcacc ctgcgagtgg
     2581 agctgctggg ctgtggtaag cttagatctt gcactgaacc cctaggcctg aaggataata
     2641 ccatccccaa caagcagatc acagcctcca gctactacaa aacctggggc ctgagtgcct
     2701 ttagctggtt tccctactac gcacgactgg ataatcaggg caagttcaac gcctggaccg
     2761 cccagaccaa cagtgcctct gagtggctgc agattgacct gggctcccag aagcgagtca
     2821 cgggcatcat cacccagggt gcccgagact ttggccacat tcaatatgtg gctgcctaca
     2881 gggtggccta tggtgatgat ggtgtgacct ggactgagta caaggacccg ggggcctcag
     2941 aaagcaagat tttccctggt aacatggaca ataattccca caagaagaac atatttgaga
     3001 cgccgttcca ggctcgcttc gtgcggatcc agcccgtggc ctggcacaac cgtatcaccc
     3061 tgcgagtgga gctgctgggc tgttaggaat tctgcagtcg acggtaccgc gggcccggga
     3121 tccaccggat ctagataact gatcataatc agccatacca catttgtaga ggttttactt
     3181 gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt
     3241 gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     3301 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     3361 gtatcttaac gcgtaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg
     3421 ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa
     3481 agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa
     3541 gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg
     3601 tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa
     3661 ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa
     3721 ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct
     3781 gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac
     3841 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     3901 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     3961 tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     4021 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     4081 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     4141 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     4201 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg
     4261 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     4321 gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc
     4381 aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat
     4441 cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt
     4501 caagaccgac ctgtccggtg ccctgaatga actgcaagac gaggcagcgc ggctatcgtg
     4561 gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag
     4621 ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc
     4681 tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc
     4741 tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga
     4801 agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga
     4861 actgttcgcc aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg
     4921 cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg
     4981 tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc
     5041 tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc
     5101 cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg
     5161 gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc
     5221 gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc
     5281 ctccagcgcg gggatctcat gctggagttc ttcgcccacc ctagggggag gctaactgaa
     5341 acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat
     5401 aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac
     5461 tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc
     5521 ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca
     5581 ggccctgcca tagcctcagg ttactcatat atactttaga ttgatttaaa acttcatttt
     5641 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     5701 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     5761 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     5821 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     5881 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     5941 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     6001 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     6061 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     6121 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     6181 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     6241 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     6301 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     6361 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     6421 tcccctgatt ctgtggataa ccgtattacc gccatgcatt agttattaat agtaatcaat
     6481 tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa
     6541 tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt
     6601 tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta
     6661 aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt
     6721 caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc
     6781 tacttggcag tacatctacg tattagtcat cgctattacc atg
//
