LOCUS       sequence-245334-        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"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      607..616
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(613..615,616..618,619..1329)
                     /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(658..679)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(919..938)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1266..1287
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1937..1955)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2205..2224
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    3611..3732
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3648..3667)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3702..3721
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3739..4194)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3876..3897)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4088..4107
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4221..4325
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4327..4684
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4327..4518
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4350..4370)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4535..4670
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4719..5513
                     /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             4719..5513
                     /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(4773..4792)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5383..5402
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5701..5720)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5745..5792
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6121..6709
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6610..6629
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
      661 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
      721 gatgccacct acggcaagct gaccctgaag ctgatctgca ccaccggcaa gctgcccgtg
      781 ccctggccca ccctcgtgac caccctgggc tacggcgtgc agtgcttcgc ccgctacccc
      841 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
      901 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
      961 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     1021 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat caccgccgac
     1081 aagcagaaga acggcatcaa ggccaacttc aagatccgcc acaacatcga ggacggcggc
     1141 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1201 cccgacaacc actacctgag ctaccagtcc aagctgagca aagaccccaa cgagaagcgc
     1261 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     1321 ctgtacaagt ccggactcag atctacgatg gcatcttctg atattcaggt gaaagagctg
     1381 gagaagcgcg cttcaggcca ggcttttgag ctgattctca gccctcggtc aaaagaatct
     1441 gtccccgatt tccccctttc ccccccaaag aagaaggacc tttccctgga ggaaatccag
     1501 aagaaattag aagctgcaga agaaagacgc aagtctcatg aggcggaagt cttgaagcag
     1561 ctcgcggaga agcgggagca tgagaaggag gtgctccaga aagccatcga ggagaacaac
     1621 aacttcagca agatggcgga ggagaagctg acccacaaaa tggaggctaa caaagagaac
     1681 cgggaggcgc agatggcggc caagctggag cgcttgcgag agaaggacaa gcacgtggaa
     1741 gaggtgcgga agaacaaaga atccaaagac cccgcggatg agactgaggc tgacggatcc
     1801 ggttcagtga gcaagggcga ggcagtgatc aaggagttca tgcggttcaa ggtgcacatg
     1861 gagggctcca tgaacggcca cgagttcgag atcgagggcg agggcgaggg ccgcccctac
     1921 gagggcaccc agaccgccaa gctgaaggtg accaagggtg gccccctgcc cttctcctgg
     1981 gacatcctgt cccctcagtt catgtacggc tccagggcct tcatcaagca ccccgccgac
     2041 atccccgact actataagca gtccttcccc gagggcttca agtgggagcg cgtgatgaac
     2101 ttcgaggacg gcggcgccgt gaccgtgacc caggacacct ccctggagga cggcaccctg
     2161 atctacaagg tgaagctcat cggcaccaac ttccctcctg acggccccgt aatgcagaag
     2221 aagacaatgg gctgggaagc gtccaccgag cggttgtacc ccgaggacgg cgtgctgaag
     2281 ggcgacatta agatggccct gcgcctgaag gacggcggcc gctacctggc ggacttcaag
     2341 accacctaca aggccaagaa gcccgtgcag atgcccggcg cctacaacgt cgaccgcaag
     2401 ttggacatca cctcccacaa cgaggactac accgtggtgg aacagtacga acgctccgag
     2461 ggccgccact ccaccggaag tagttgcact gaacccctag gcctgaagga taataccatc
     2521 cccaacaagc agatcacagc ctccagctac tacaaaacct ggggcctgag tgcctttagc
     2581 tggtttccct actacgcacg actggataat cagggcaagt tcaacgcctg gaccgcccag
     2641 accaacagtg cctctgagtg gctgcagatt gacctgggct cccagaagcg agtcacgggc
     2701 atcatcaccc agggtgcccg agactttggc cacattcaat atgtggctgc ctacagggtg
     2761 gcctatggtg atgatggtgt gacctggact gagtacaagg acccgggggc ctcagaaagc
     2821 aagattttcc ctggtaacat ggacaataat tcccacaaga agaacatatt tgagacgccg
     2881 ttccaggctc gcttcgtgcg gatccagccc gtggcctggc acaaccgtat caccctgcga
     2941 gtggagctgc tgggctgtgg taagcttaga tcttgcactg aacccctagg cctgaaggat
     3001 aataccatcc ccaacaagca gatcacagcc tccagctact acaaaacctg gggcctgagt
     3061 gcctttagct ggtttcccta ctacgcacga ctggataatc agggcaagtt caacgcctgg
     3121 accgcccaga ccaacagtgc ctctgagtgg ctgcagattg acctgggctc ccagaagcga
     3181 gtcacgggca tcatcaccca gggtgcccga gactttggcc acattcaata tgtggctgcc
     3241 tacagggtgg cctatggtga tgatggtgtg acctggactg agtacaagga cccgggggcc
     3301 tcagaaagca agattttccc tggtaacatg gacaataatt cccacaagaa gaacatattt
     3361 gagacgccgt tccaggctcg cttcgtgcgg atccagcccg tggcctggca caaccgtatc
     3421 accctgcgag tggagctgct gggctgttag gaattctgca gtcgacggta ccgcgggccc
     3481 gggatccacc ggatctagat aactgatcat aatcagccat accacatttg tagaggtttt
     3541 acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat
     3601 tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     3661 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     3721 caatgtatct taacgcgtaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt
     3781 tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat
     3841 caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat
     3901 taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac
     3961 tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc
     4021 ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga
     4081 gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca
     4141 cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg
     4201 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     4261 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     4321 agagtcctga ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc
     4381 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     4441 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     4501 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     4561 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     4621 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     4681 caaagatcga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
     4741 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     4801 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     4861 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca agacgaggca gcgcggctat
     4921 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     4981 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     5041 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     5101 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     5161 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     5221 ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc
     5281 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     5341 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     5401 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     5461 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
     5521 tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     5581 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     5641 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccctaggg ggaggctaac
     5701 tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca
     5761 gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg
     5821 gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt
     5881 ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc
     5941 ggcaggccct gccatagcct caggttactc atatatactt tagattgatt taaaacttca
     6001 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     6061 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     6121 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     6181 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     6241 cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt
     6301 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     6361 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     6421 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     6481 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     6541 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     6601 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     6661 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     6721 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     6781 gttatcccct gattctgtgg ataaccgtat taccgccatg cat
//
