LOCUS       sequence-209380-        5155 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  C-terminal half of pi-EB1, mCherry-tagged, dissociates from 
            microtubule ends in blue light.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5155)
  AUTHORS   van Haren J, Charafeddine RA, Ettinger A, Wang H, Hahn KM, Wittmann 
            T
  TITLE     Local control of intracellular microtubule dynamics by EB1 
            photodissociation.
  JOURNAL   Nat Cell Biol. 2018 Jan 29. pii: 10.1038/s41556-017-0028-5. doi: 
            10.1038/s41556-017-0028-5.
  PUBMED    29379139
REFERENCE   2  (bases 1 to 5155)
  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: "Nat Cell 
            Biol. 2018 Jan 29. pii: 10.1038/s41556-017-0028-5. doi: 
            10.1038/s41556-017-0028-5."
COMMENT     Sequence Label: pmCherry-Zdk1-EB1C
FEATURES             Location/Qualifiers
     source          1..5155
                     /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"
     CDS             600..1307
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(745..763)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     CDS             1320..1496
                     /codon_start=1
                     /product="""Zdark"" protein, derived from the Z subunit of 
                     protein A, that binds to the dark state of LOV2 and is 
                     released upon irradiation (Wang et al., 2016)"
                     /label=Zdk1
                     /translation="MVDNKFNKEKTRAGAEIHSLPNLNVEQKFAFIVSLFDDPSQSANL
                     LAEAKKLNDAQAPK"
     misc_feature    1767..1819
                     /label=MCS
                     /note="multiple cloning site"
     polyA_signal    1943..2064
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1980..1999)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2034..2053
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2071..2526)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2208..2229)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2420..2439
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2553..2657
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2659..3016
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2659..2850
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2682..2702)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2867..3002
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3051..3845
                     /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             3051..3845
                     /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(3105..3124)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3715..3734
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4033..4052)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4077..4124
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4453..5041
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4942..4961
                     /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 gctagcacca
      601 tggtgagcaa gggcgaggag gataacatgg ccatcatcaa ggagttcatg cgcttcaagg
      661 tgcacatgga gggctccgtg aacggccacg agttcgagat cgagggcgag ggcgagggcc
      721 gcccctacga gggcacccag accgccaagc tgaaggtgac caagggtggc cccctgccct
      781 tcgcctggga catcctgtcc cctcagttca tgtacggctc caaggcctac gtgaagcacc
      841 ccgccgacat ccccgactac ttgaagctgt ccttccccga gggcttcaag tgggagcgcg
      901 tgatgaactt cgaggacggc ggcgtggtga ccgtgaccca ggactcctcc ctgcaggacg
      961 gcgagttcat ctacaaggtg aagctgcgcg gcaccaactt cccctccgac ggccctgtaa
     1021 tgcagaagaa gactatgggc tgggaggcct cctccgagcg gatgtacccc gaggacggcg
     1081 ccctgaaggg cgagatcaag cagaggctga agctgaagga cggcggccac tacgacgctg
     1141 aggtcaagac cacctacaag gccaagaagc ccgtgcagct gcccggcgcc tacaacgtca
     1201 acatcaagtt ggacatcacc tcccacaacg aggactacac catcgtggaa cagtacgaac
     1261 gcgccgaggg ccgccactcc accggcggca tggacgagct gtacaagggt tctggatcca
     1321 tggtggataa caaattcaat aaagaaaaga cgcgtgccgg tgcggaaatc cattctctgc
     1381 caaacctgaa tgttgagcag aagtttgcct tcatcgtgag cctgtttgat gatccatctc
     1441 agagcgccaa tctgctggcc gaagccaaaa aactgaacga tgcccaggcc ccaaaaggtg
     1501 gttctggtgg tagcgtgggc aacggagacg acgaggcagc tgagttgatg cagcaggtca
     1561 acgtattgaa acttactgtt gaagacttgg agaaagagag ggatttctac ttcggaaagc
     1621 tacggaacat tgaattgatt tgccaggaga acgaggggga aaacgaccct gtattgcaga
     1681 ggattgtaga cattctgtat gccacagatg aaggctttgt gatacctgat gaagggggcc
     1741 cacaggagga gcaagaagag tattaactcg agctcaagct tcgaattctg cagtcgacgg
     1801 taccgcgggc ccgggatcca ccggatctag ataactgatc ataatcagcc ataccacatt
     1861 tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa
     1921 aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag
     1981 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     2041 gtccaaactc atcaatgtat cttaacgcgt aaattgtaag cgttaatatt ttgttaaaat
     2101 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa
     2161 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca
     2221 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
     2281 gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta
     2341 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg
     2401 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa
     2461 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
     2521 gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     2581 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     2641 attgaaaaag gaagagtcct gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg
     2701 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     2761 gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     2821 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac
     2881 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     2941 ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     3001 cctaggcttt tgcaaagatc gatcaagaga caggatgagg atcgtttcgc atgattgaac
     3061 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
     3121 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
     3181 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caagacgagg
     3241 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg
     3301 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt
     3361 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc
     3421 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag
     3481 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg
     3541 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc
     3601 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt
     3661 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg
     3721 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt
     3781 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct
     3841 tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg
     3901 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
     3961 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccctag
     4021 ggggaggcta actgaaacac ggaaggagac aataccggaa ggaacccgcg ctatgacggc
     4081 aataaaaaga cagaataaaa cgcacggtgt tgggtcgttt gttcataaac gcggggttcg
     4141 gtcccagggc tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg
     4201 cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc
     4261 caacgtcggg gcggcaggcc ctgccatagc ctcaggttac tcatatatac tttagattga
     4321 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     4381 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     4441 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     4501 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     4561 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     4621 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     4681 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     4741 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     4801 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     4861 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     4921 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     4981 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     5041 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     5101 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcca tgcat
//
